The materials and methods are briefly summarized here; expanded materials and methods are included in the supplementary materials . Data were generated across human and microbial cells, including isolations of stool, saliva, skin, urine, blood, plasma, PBMCs, and immune cells that are CD4+, CD8+, and CD19+ enriched and lymphocyte-depleted (LD), from AutoMACS magnetic bead separation and validated by FACS (fig. S1 ). Molecular techniques included assessments of telomere length, telomerase activity, and chromosome aberration frequencies (qRT-PCR T:A, qRT-PCR TRAP, Telo-FISH, and dGH), WGBS, RNA-seq (polyA, riboRNA, and miRNA), mitochondrial quantification (qPCR and qRT-PCR), shotgun metagenome sequencing of fecal microbiome, targeted proteomics (LC-MS), untargeted proteomics (PECAN, MaxQuant for urine and SWATH-MS for plasma), targeted metabolomics (GC-MS), untargeted metabolomics (LC-MS), mitochondrial respiration (Seahorse XF), oxidative state measures (EPR), TCR and BCR (T cell and B cell receptor repertoire) profiling, 10 cognitive tests (motor praxis, visual object learning, fractal 2-back, abstract matching, line orientation, emotion recognition, matrix reasoning, digit symbol substitution, balloon analog risk, and psychomotor vigilance), vascular and ocular measures by ultrasound and optical coherence tomography, respectively, and a wide range of other biometrics (e.g., nutrition, height, and weight). Finally, a large set of biochemical profiles were measured pre-, in-, and postflight for both subjects: body mass, height, energy intake, vitamin levels (A, B6, B12, C, D, and E and 1-carbon metabolites), minerals (copper, ceruloplasmin, selenium, zinc, calcium, phosphorus, magnesium, and iodine), iron levels (ferritin, transferrin, transferrin receptors, Hgb, Hct, MCV, TIBC, and hepcidin), urine proteins (total, albumin, TTR, RBP, creatinine, metallothionein, 3-MH, nitrogen, and fibrinogen), bone markers (BSAP, PTH, OPG, RANKL, P1NP, sclerostin, and osteocalcin), collagen crosslinks (NTX, CTX, and DPD), oxidative stress and antioxidant capacity (8-OHdG, PGF2α, GPX, SOD, TAC, oxLDL, total lipid peroxides, heme, and glutathione), protein carbonyls (myeloperoxidase, lp-PLA2, neopterin, and beta-2 microglobulin), hormones and immune system markers (cytokines, testosterone, estradiol, DHEA/S, cortisol, IGF1, leptin, thyroid hormones, angiotensin, aldosterone, ANP, PRA, and insulin), and general urine chemistry (Na, K, and Cl ions; uric acid; cholesterol; triglyceride; HDL; LDL; phospholipids; renal stone risk; liver enzymes; hsCRP; NAD/P; and pH). Together, these data span 25 months for the flight subject twin (TW), who was compared with himself, either preflight, inflight, or postflight, and also with his twin control (HR) on Earth using generalized linear models (GLM), DESeq2, and fuzzy c-means clustering for longitudinal trends. All P values were corrected for multiple testing using a FDR of 0.05 or 0.01, and q values are reported in all tables.
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Leptin receptor, human
Leptin receptor, human
The leptin receptor is a cell surface receptor that binds to the hormone leptin.
Leptin is an important regulator of energy balance, food intake, and body weight.
The leptin receptor is expressed in various tissues, including the hypothalamus, where it mediates leptin's effects on appetite and metabolism.
Proper functioning of the leptin receptor is crucial for maintaining normal body weight and energy homeostasis.
Disfunctions in the leptin recepotr pathway have been implicated in obesity and related metabolic disorders.
Leptin is an important regulator of energy balance, food intake, and body weight.
The leptin receptor is expressed in various tissues, including the hypothalamus, where it mediates leptin's effects on appetite and metabolism.
Proper functioning of the leptin receptor is crucial for maintaining normal body weight and energy homeostasis.
Disfunctions in the leptin recepotr pathway have been implicated in obesity and related metabolic disorders.
Most cited protocols related to «Leptin receptor, human»
8-Hydroxy-2'-Deoxyguanosine
Albumins
Aldosterone
Angiotensins
Antioxidants
BETA MICROGLOBULIN 2
BLOOD
Blood Vessel
Bones
Calcium
Carbon
Cell Respiration
Cells
Ceruloplasmin
Cholesterol
Chromosome Aberrations
Cognitive Testing
Collagen
Copper
C Reactive Protein
Creatinine
Cytokine
Dehydroepiandrosterone Sulfate
Dinoprost
Emotions
Enzymes
Estradiol
Feces
Ferritin
Fibrinogen
Fingers
Fishes
Gas Chromatography-Mass Spectrometry
Glutathione
Heme
Hepcidin
Homo sapiens
Hormones
Human Body
Hydrocortisone
IGF1 protein, human
Insulin
Iodine
Ions
Iron
isolation
Kidney Calculi
Leptin
Lipid Peroxides
Liver
Lymphocyte
Magnesium
Metagenome
Metallothionein
Microbiome
MicroRNAs
Minerals
Mitochondria
Neopterin
Nitrogen
Osteocalcin
Oxidative Stress
oxidized low density lipoprotein
PAF 2-Acylhydrolase
PAX5 protein, human
Pecans
Peroxidase
Phospholipids
Phosphorus
Plasma
Poly A
procollagen Type I N-terminal peptide
Proteins
Receptors, Antigen, B-Cell
RNA-Seq
Saliva
Seahorses
Selenium
Skin
T-Lymphocyte
Telomerase
Telomere
Testosterone
Thyroid Hormones
TNFSF11 protein, human
Tomography, Optical Coherence
Transferrin
Transferrin Receptor
Triglycerides
Twins
Ultrasonics
Uric Acid
Urine
Vision
Vitamins
Wakefulness
Zinc
3-benzoyl dopamine
Biological Markers
CHI3L1 protein, human
Epidermal growth factor
Interleukin-6
Interstitial Collagenase
Leptin
liquid crystal polymer
Matrix Metalloproteinase 3
Resistin
TNFRSF1A protein, human
Vascular Cell Adhesion Molecule-1
VEGF protein, human
positional cloning approach and the comparative transfer approach were used to target various
gene families for discovery of gene networks associated with economically important traits in
beef cattle. A total of 71 genes that had mutations detected and genotyped successfully in our
beef reference population (see Animals, Phenotypes and Statistical Analysis) are illustrated in
Figure
classified into five gene families.
The first family involves nuclear encoded mitochondrial genes, such as aldehyde dehydrogenase
4 family, member A1 (ALDH4A1), amyloid beta (A4) precursor protein
(APP), ATP synthase, H+ transporting, mitochondrial F1 complex, O subunit
(ATP5O), BCL2-antagonist/killer 1 (BAK1), chromosome 21 open
reading frame 2 (C21orf2), collagen, type VI, alpha 1
(COL6A1), C-reactive protein, pentraxin-related (CRP),
enhancer of yellow 2 homolog (Drosophila) (ENY2), fatty acid binding protein 3
(FABP3) 5 (link), fatty acid binding protein 4
(FABP4) 6 (link), mitochondrial fission
regulator 1 (MTFR1), mitochondrial ribosomal protein L39
(MRPL39), polymerase (RNA) mitochondrial (DNA directed)
(POLRMT), Poly (A) polymerase associated domain containing 1
(PAPD1) 7 (link), RAB2A, member RAS oncogene
family (RAB2A), regulator of calcineurin 1 (RCAN1),
single-minded homolog 2 (Drosophila) (SIM2), superkiller viralicidic activity
2-like (S. cerevisiae) (SKIV2L), transcription factor A, mitochondrial
(TFAM) 8 (link), transcription factor B1,
mitochondrial (TFB1M), transcription factor B2, mitochondrial
(TFB2M), tumor necrosis factor (TNF superfamily, member 2)
(TNF), ubiquinol-cytochrome c reductase core protein I
(UQCRC1) 9 (link) and uncoupling protein 1
(UCP1).
The second family is related to the long chain fatty acids uptake gene complex, including
solute carrier family 2, member 2 (SLC2A2), solute carrier family 25, member 27
(SLC25A27) and solute carrier family 27, member 1 (SLC27A1),
member 2 (SLC27A2) and member 4 (SLC27A4).
The third family deals with the sauvagine/corticotropin-releasing factor/urotensin I family
and related families, such as corticotropin releasing hormone (CRH) 10 (link), CRH receptor 1 (CRHR1), CRH receptor 2
(CRHR2) 11 (link), urocortin 3
(UCN3), urotensin 2 (UTS2) and urotensin 2 receptor
(UTS2R) 12 (link).
The fourth family targets the lipogenesis/lipolysis enzymes, such as acetyl-Coenzyme A
acetyltransferase 2 (ACAT2), acyl-CoA synthetase long-chain family member 5
(ACSL5), 7-dehydrocholesterol reductase (DHCR7),
diacylglycerol O-acyltransferase homolog 1 (DGAT1) 5 (link), fibronectin type III domain containing 3B (FNDC3B),
3-hydroxy-3-methylglutaryl-Coenzyme A synthase 1 (soluble) (HMGCS1),
3-hydroxymethyl-3-methylglutaryl-Coenzyme A lyase (HMGCL), lipase,
hormone-sensitive (LIPE), patatin-like phospholipase domain containing 2
(PNPLA2), stearoyl-CoA desaturase (delta-9-desaturase) (SCD1)
13 (link) and sterol O-acyltransferase 1
(SOAT1).
The fifth family focuses on calpain/calpasatin or related genes, such as calpain 1
(CAPN1), calpain 3 (CAPN3), calpain 5
(CAPN5), calpain 7 (CAPN7), calpain 8
(CAPN8), calpain 9 (CAPN9), calpain 11
(CAPN11), calpain 12 (CAPN12), calpain 14
(CAPN14), calpain, small subunit 1 (CAPNS1), calpastatin
(CAST) 14 (link), dermatopontin
(DPT), neuromedin U (NMU), troponin I type 2 (skeletal, fast)
(TNNI2) and troponin T type 1 (skeletal, slow) (TNNT1).
In addition, ankyrin repeat and SOCS box-containing 3 (ASB3), chromodomain
helicase DNA binding protein 9 (CHD9), dopey family member 2
(DOPEY2), epidermal growth factor receptor pathway substrate 15
(EPS15), growth hormone 1 (GH1), histone cluster 1, H1t
(HIST1H1T), leptin (LEP), proteasome (prosome, macropain)
assembly chaperone 1 (PSMG1), thyroglobulin (TG) and tRNA
nucleotidyl transferase, CCA-adding, 1 (TRNT1) were also investigated in the
present study.
sequences for the candidate genes described above. We used cDNA sequences of the human orthologs
as references for BLAST searches to retrieve the orthologous cDNA sequences against the GenBank
database “nr” or the orthologous ESTs sequences against the GenBank database
“est_others” with a species option limited to Bos taurus.
The cDNA sequences in the “nr” database represent three categories: cDNA
sequences derived from a full-length cDNA library, known gene cDNA sequences or annotated cDNA
sequences compiled by the GenBank staff. We collected the longest cDNA sequence retrieved from
the “nr” database or a cDNA sequence assembled from several ESTs retrieved
from the “est_others” database to form a primary cDNA sequence for each
cattle gene. This sequence was then used to perform a species-specific BLAST search against the
“est_others” database in order to expand the primary sequence to a
full-length cDNA sequence. At the end, we used the full-length cDNA sequence to search for
genomic DNA contigs in the 7.15X bovine genome sequence database (see the Bovine Genome
Resources at NCBI). The cDNA sequences and genomic DNA sequences were aligned to
determine the genomic organizations of all genes investigated in the present study.
The online oligonucleotide design tool Primer3 (
following criteria: 18-25 bp in length, >50% in GC content and optimal Tm of either
60oC or 65oC. We mainly targeted the promoter region and the
3'untranslated region (UTR) of each gene to maximize the chance for mutation detection. The
sizes of the amplified products for most amplicons ranged from 400 - 600 bp, which is a
sufficient length for accurate sequencing analysis. PCR reactions were performed using 25 ng of
bovine genomic DNA as template in a final volume of 10 μL containing 12.5 ng of each
primer, 200 μM dNTPs, 1.5 - 3 mM MgCl2, 50 mM KCl, 20 mM Tris-HCl and 0.2U
of Platinum Taq polymerase (Invitrogen, Carlsbad, CA). The PCR conditions were carried out as
follows: 94oC for 2 min, 32 cycles of 94oC for 30 sec, 61oC for
30 sec and 72oC for 30 sec, followed by a further 5 min extension at 72oC.
PCR products were examined by electrophoresis through a 1.5% agarose gel with 1X TBE buffer to
determine the quality and quantity for DNA sequencing. Sequencing was performed on ABI 3730
sequencer in the Laboratory for Biotechnology and Bioanalysis (Washington State University).
Mutations were identified using six Wagyu-Limousin F1 animals (see animals below) and
138 of them were successfully genotyped on all animals using a Sequenom iPLEX assay service
provided by the Children's Hospital Oakland Research Institute, Oakland, California.
including 6 F1 bull, 113 F1 dams and 246 F2 progeny. We focused
on a total of 19 phenotypic measurements, which can be classified into three categories: carcass
measurements, including carcass weight (CW), ribeye area (REA), subcutaneous fat depth (SFD),
percentage of kidney, pelvic and heart fat (KPH) and beef marbling scores (BMS); eating quality,
including shear force of cooked steak (SFCS), taste panel myofibrillar tenderness (TPMT), taste
panel connective tissue (TPCT) content, taste panel overall tenderness (TPOT) rating, taste
panel juiciness (TPJN), taste panel flavor evaluation (TPFE); and fatty acid composition
including three indexes of Δ9 desaturase activity - R1 = 14:1 to
14:0, R2 = 16:1 to 16:0 and R3 = 18:1 to 18:0, relative amounts of
saturated (SFA), monounsaturated (MUFA) and poly unsaturated fatty acids (PUFA), conjugated
linoleic acid/100 g dry meat (CLA) and cholesterol/100 g dry meat (CHOL). Development/management
of the Wagyu-Limousin reference population and measurement/definition of these phenotypes were
described previously 8 (link),15 (link)-18 .
The HAPLOVIEW program 19 (link) was used to determine the
linkage disequilibrium (LD) of 138 markers located on 22 bovine chromosomes, thus leading to
selection of tag mutations for association analysis. The association between genotypes and
traits was evaluated using the general linear model (GLM) procedure of SPSS (version 16.0) (The
Predictive Analytic Company, Chicago, USA). The model was:
where yijklm is phenotypic observation of a quantitative trait for
animal m, sexi is the effect of the i-th sex
category (i=1,2), yearj is the effect of the j-th
harvest year (j=1,2), age is a covariate for age in days of
the animal at harvest, snpk represents the effects of each genotype
at the k-th SNP locus, and sirel is random effect of the l-th sire
producing animal m, and eijklm is a residual term
pertaining to animal m. In the model, we assumed that where is the variance of
sire effects, and A is an additive genetic relationship matrix among the sires, and
where
is
the residual variance. If the effects of sire, sex, year, or age were not significant
(P>0.05) after initial analysis, they were removed from the model for final analysis.
All single marker-trait associations that reached a significance level of P<0.05 were
initially included in further analysis. We discarded significant markers when there were 9 or
fewer animals in one genotype group or there were only two genotypes rather than three. Based on
the pairwise significance tests among three genotypes, we classified the remaining significant
associations into three quantitative trait modes (QTMs): 1) additive mode when PAa ≈ (PAA + Paa)/2; 2) dominant mode when PAa ≈ either PAA or Paa; and 3) overdominant mode when
PAa > or < both PAA and Paa, where
PAa = least square means of heterozygous animals, PAA = least square means
of homozygous animals with higher performance and Paa = least square means of
homozygous animals with lower performance. We then integrated these markers along with their
QTMs into a linear regression analysis using the linear regression procedure (SPSS for Windows,
version 16.0) in order to identify gene-gene combinations, i.e., gene-networks related to
carcass traits, eating quality and fatty acid composition in beef.
Protein extracts (100 μg) were subjected to immunoprecipitation using a protein A/G agarose column (Amicogen, Korea). Protein A/G agarose columns were separately pre-incubated with 1 μg of 216 different antisera for; growth and proliferation-related proteins (n = 10), cMyc/MAX/MAD signaling proteins (n = 3), p53/Rb/E2F signaling proteins (n = 5), epigenetic modification-related proteins (n = 6), protein translation-related proteins (n = 5), RAS signaling proteins (n = 17), growth factor-related proteins (n = 16), NFkB signaling proteins (n = 12), cellular protection-related proteins (n = 15), upregulated inflammatory proteins (n = 26), downregulated inflammatory proteins (n = 13), p53-mediated apoptosis-related proteins (n = 17), FAS-mediated apoptosis-related proteins (n = 8), angiogenesis-related proteins (n = 20), osteogenesis-related proteins (n = 12), antioxidant-related proteins (n = 8), oncogenic proteins (n = 15), and control housekeeping proteins (n = 3) (Table 2 ).
2 ). Protein percentages in total proteins in experimental and control groups were plotted. Analyses were repeated two to six times to achieve mean standard deviations of ≤±5%. Results were analyzed using the Chi-squared test.
The expressions of housekeeping proteins, that is, β-actin, α-tubulin, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were used as internal controls. Expressional changes of housekeeping proteins were adjusted to <±5% using a proportional basal line algorithm. To describe protein expressional changes, we tentatively defined a ≤±5% change as minimal, ±5–10% as slight, ±10–20% as meaningful, and a ≥±20% change as marked.
Antibodies used in the study.
| Signaling proteins | No. | Antibodies |
|---|---|---|
| Cellular proliferation | 10 | Ki-67 |
| cMyc/MAX/MAD signaling | 3 | cMyc*, MAX |
| p53/Rb/E2F signaling | 5 (2) | p53, Rb-1#, E2F-1*, (p21, CDK4) |
| Wnt/β-catenin signaling | 5 | Wnt1*, β-catenin*, APC*, snail*, TCF-1* |
| Epigenetic modification | 6 | DMAP1 |
| Protein translation signaling | 5 | DOHH |
| RAS signaling | 17 | NRAS$, KRAS$, STAT3*, SOS-1 |
| Growth factor signaling | 16 | FGF-1 |
| NFkB signaling | 12 (3) | NFkB |
| Upregulated inflammatory proteins | 26 (2) | IL-12 |
| Downregulated inflammatory proteins | 13 (1) | TNFα@, IL-1 |
| Cellular protection-related | 15 (2) | LC3, PLC- β2, PI3K, PKC |
| Antioxidant-related | 8 (3) | HO-1 |
| p53-mediated cellular apoptosis | 17 (1) | (p53*), PUMA |
| FAS-mediated cellular apoptosis | 8 (3) | FASL |
| Oncogenic proteins | 15 (2) | PTEN&, MUC1, MUC4, maspin*, BRCA1&, BRCA2&, NF-1 |
| Angiogenesis-related proteins | 20 (7) | HIF&, VEGF-A |
| Osteogenesis-related proteins | 12 (2) | OPG |
| Control housekeeping proteins | 3 | α-tubulin*, β-actin |
| Total | 216 (28) |
*Santa Cruz Biotechnology, USA; #DAKO, Denmark; $Neomarkers, CA, USA; @ZYMED, CA, USA; &Abcam, Cambridge, UK; the number of antibodies overlapped; ().
Abbreviations: AMPK; AMP-activated protein kinase, pAKT; v-akt murine thymoma viral oncogene homolog, p-Akt1/2/3 phosphorylated (p-Akt, Thr 308), APAF-1; apoptotic protease-activating factor 1, AP-1; activating protein-1, BAD; BCL2 associated death promoter, BAK; BCL2 antagonist/killer, BAX; BCL2 associated X, BCL-2; B-cell leukemia/lymphoma-2, BID; BH3 interacting-domain death agonist, c-caspase 3; cleaved-caspase 3, CD3; cluster of differentiation 3, CDK4; cyclin dependent kinase 4, CEA; carcinoembryonic antigen, CMG2: capillary morphogenesis protein 2, COX-1; cyclooxygenase-2, COX-2; cyclooxygenase-2, c-PARP; cleaved- PARP (poly-ADP ribose polymerase), DMAP1; DNA methyltransferase 1 associated protein, DMBT1; deleted in malignant brain tumors 1, DOHH; deoxyhypusine hydroxylase, DHS; deoxyhypusine synthase, E2F-1; transcription factor, eIF2AK3 (PERK); eukaryotic translation initiation factor 2 (protein kinase R (PKR)-like endoplasmic reticulum kinase), elF5A-1; eukaryotic translation initiation factor 5A-1, elF5A-2; eukaryotic translation initiation factor 5A-2, ERβ; estrogen receptor beta, ERK; extracellular signal-regulated protein kinases, ET-1: endothelin-1, FAS; CD95/Apo1, FASL; FAS ligand, FADD; FAS associated via death domain, FGF-1; fibroblast growth factor-1, FLIP; FLICE-like inhibitory protein, FLT-4; Fms-related tyrosine kinase 4, GADD45; growth arrest and DNA-damage-inducible 45, GAPDH; glyceraldehyde 3-phosphate dehydrogenase, GH; growth hormone, GHRH; growth hormone-releasing hormone, GST-1; glutathione S-transferase ω 1, HDAC-10; histone deacetylase 10, HIF-1α: hypoxia inducible factor-1α, HO-1; heme oxygenase 1, HER1; human epidermal growth factor receptor 1, HGFα; hepatocyte growth factor α, HSP-70; heat shock protein-70, IKK; ikappaB kinase, IGF-1; insulin-like growth factor 1, IGFIIR; insulin-like growth factor 2 receptor, IgK; immunoglobulin kappa (light chain), IL-1; interleukin-1, KDM4D; Lysine-specific demethylase 4D, JNK-1; Jun N-terminal protein kinase, KRAS; V-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog, LC3; microtubule-associated protein 1 A/1B-light chain 3, LYVE-1: lymphatic vessel endothelial hyaluronan receptor 1, MAX; myc-associated factor X, MBD4; methyl-CpG-binding domain protein 4, M-CSF; macrophage colony-stimulating factor, MDM2; mouse double minute 2 homolog, MDR; multiple drug resistance, MMP-1; matrix metalloprotease-1, MPM2; mitotic protein monoclonal 2, mTOR; mammalian target of rapamycin, cMyc; V-myc myelocytomatosis viral oncogene homolog, NFkB; nuclear factor kappa-light-chain-enhancer of activated B cells, NOS-1; nitric oxide synthase 1, NRAS; neuroblastoma RAS Viral Oncogene homolog, NRF2; nuclear factor (erythroid-derived)-like 2, p14, p16, p21, p27, p38, PAI-1; plasminogen activator inhibitor-1, PARP; poly-ADP ribose polymerase, PCNA; proliferating cell nuclear antigen, PDGF-A: platelet-derived growth factor-A, PLC-β2; 1-phosphatidylinositol-4,5-bisphosphate phosphodiesterse β-2, PI3K; phosphatidylinositol-3-kinase, PLK4; polo like kinase 4 or serine/threonine-protein kinase, PKC; protein kinase C, p-p38; phosphor-p38, PTEN; phosphatase and tensin homolog, RANKL; receptor activator of nuclear factor kappa-B ligand, Rb-1; retinoblastoma-1, RUNX2; Runt-related transcription factor-2, SMAD4; mothers against decapentaplegic, drosophila homolog 4, SOD-1; superoxide dismutase-1, SP-1; specificity protein 1, STAT3; signal transducer and activator of transcription-3, TGF-β1; transforming growth factor-β1, TERT; human telomerase reverse transcriptase, TNFα; tumor necrosis factor-α, β-actin, 14-3-3, VEGF vascular endothelial growth factor, VEGFR2: vascular endothelial growth factor receptor 2, p-VEGFR2: vascular endothelial growth factor receptor 2 (Y951), vWF: von Willebrand factor.
The expressions of housekeeping proteins, that is, β-actin, α-tubulin, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were used as internal controls. Expressional changes of housekeeping proteins were adjusted to <±5% using a proportional basal line algorithm. To describe protein expressional changes, we tentatively defined a ≤±5% change as minimal, ±5–10% as slight, ±10–20% as meaningful, and a ≥±20% change as marked.
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Most recents protocols related to «Leptin receptor, human»
The human leptin receptor ELISA kit (ab282876) was used to quantitate the levels of the human leptin receptor protein in cell extracts following the manufacturer’s instructions. Briefly, the assay employs capture antibodies conjugated to an affinity tag that is detected by a monoclonal antibody used to coat the ELISA wells, allowing formation of the antibody-analyte sandwich complex in one step.
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IUPAC of Ghrelin is H-Gly-Ser-Ser(octanoyl)-Phe-OH. Ligand Ghrelin used Receptor 7NA8 Chain R. The entry with the identifier 7NA8 in the RCSB Protein Data Bank (PDB) represents the structures of human Ghrelin receptor-Gi complexes with Ghrelin and a synthetic agonist. Ghrelin is a peptide hormone that plays a role in regulating appetite and energy balance. It binds to the ghrelin receptor, a G proteincoupled receptor (GPCR) located on the cell membrane. The complex formed between the ghrelin receptor and Gi protein is particularly interesting because it is involved in signaling pathways regulating various physiological processes, including hunger, satiety, and metabolism. The complex includes the ghrelin receptor, a transmembrane protein, and the Gi protein, a guanine nucleotide-binding protein.
The structure provides insights into the interaction between the ghrelin receptor and its ligands, Ghrelin, and a synthetic agonist. The ligands bind to specific regions on the ghrelin receptor, triggering a cascade of signaling events that regulate cellular responses 16 . Leptin is a cytokine crucial in regulating body weight and energy balance. Mutations in the gene encoding leptin or its receptor can lead to obesity, infertility, and diabetes in mice. The crystal structure of the mutant form of human leptin (leptin-E100) was determined using Xray diffraction. The resolution of the structure is 2.40 Å, which provides detailed information about the arrangement of atoms in the protein.
The structure provides insights into the interaction between the ghrelin receptor and its ligands, Ghrelin, and a synthetic agonist. The ligands bind to specific regions on the ghrelin receptor, triggering a cascade of signaling events that regulate cellular responses 16 . Leptin is a cytokine crucial in regulating body weight and energy balance. Mutations in the gene encoding leptin or its receptor can lead to obesity, infertility, and diabetes in mice. The crystal structure of the mutant form of human leptin (leptin-E100) was determined using Xray diffraction. The resolution of the structure is 2.40 Å, which provides detailed information about the arrangement of atoms in the protein.
Serum interleukin 1 receptor antagonist (IL-1ra) concentrations were measured with the Human IL-1ra/IL-1F3 Quantikine ELISA kits (R & D Systems, Minneapolis, USA). Serum TNF (TNF-α) and IL-6 concentrations were measured with the Proinflammatory Panel 1 Human Kit (Meso Scale Diagnostics, Maryland, USA), and C-reactive protein (CRP) was analyzed with Vascular Injury Panel 1 Human Kit (Meso Scale Diagnostics, Maryland, USA). The Human Leptin, Insulin Kit (Meso Scale Diagnostics, Maryland, USA) was used to analyze serum leptin concentrations. All serum samples were tested in duplicate. The intra- and inter-assay coefficients of variation (CV) were 3.1% and 15.6% (IL-1ra), 4.2% and 7.0% (IL-6), 3.1% and 12.1% (TNF-α), 2.5% and 10.0% (CRP), and 6.6% and 8.9% (leptin), respectively.
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Venous blood samples were collected at least 2 h postprandially. Local anesthetic topical cream was used prior to sampling. The tubes were centrifuged for 10 min at 1300× g. Serum was separated into Cryo tubes and then stored at −80 °C at the Biobank in Lund or Umeå until analysis. Hemoglobin (Hb) was analyzed directly at the University and Regional Laboratories of Skåne or Norrland’s University Hospital with the Sysmex XN-10 (Sysmex Corporation, Chuoko, Japan). The tubes were transported from the Biobanks frozen on dry ice to the respective laboratory for analysis. Serum (s) insulin, C-peptide, and IGF-1 were analyzed at the University and Regional Laboratories of Skåne, insulin and C-peptide with the Cobas 601 instrument (Roche Diagnostics, Rotkreuz, Switzerland) and IGF-1 by the IDS-iSYS assay (Immunodiagnostic System Ltd., Boldon, Tyne & Wear, UK). Serum leptin and leptin receptor were analyzed by ELISA (Human Leptin ELISA kit, EMD Millipore; Merck KGaA, Darmstadt, Germany, and Human Leptin R Quantikine® ELISA, R&D Systems Inc., Minneapolis, MN, USA) at the Pediatric Research Laboratory at Umeå University.
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Most of the material used in this study was purchased as we reported earlier38 (link),55 (link)–57 (link). Phosphate Buffered Saline (PBS), nitrocellulose membranes, streptavidin-horseradish peroxidase (HRP) conjugate, Ponceau S solution, LY294002 hydrochloride, hydrogen peroxide solution, human leptin (L4146), recombinant human AChE (C1682, UniProt accession ID: C9JD78), chelerythrine chloride, and PD98059 were purchased from Sigma-Aldrich. LEP Human siRNA (AM16708), leptin mouse monoclonal antibody (MA5-23740), α-tubulin mouse monoclonal antibody (DM1A), 3,3′,5,5′-tetramethylbenzidine (TMB), Halt Protease and Phosphatase Inhibitor Cocktail, BCA protein assay kit, SuperSignal West Pico luminol (chemiluminescence) reagent, and lipofectamine 2000 transfection reagent were from ThermoFisher. Donkey anti-mouse IgG (HRP) (ab205724), goat anti-AChE antibody (ab31276), and rabbit anti-Goat IgG H&L (HRP) (ab6741) were purchased from Abcam. Anti-Human Mouse sAPPα (2B3) IgG MoAb was purchased from IBL America. SignalSilence Control siRNA (Unconjugated, 6568) was purchased from Cell Signaling Technology. Anti-Aβ mouse (6E10, 1–16) antibody, anti-Aβ42 mouse antibody that is reactive to the C-terminus of Aβ42, anti-Aβ40 mouse antibody that is reactive to the C-terminus of Aβ40, and biotin anti-Aβ mouse (6E10, 1–16) antibody were from BioLegend. Ob-R siRNA (sc-36115) and Ob-R/leptin receptor antibody (sc-8391) were purchased from Santa Cruz.
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Recombinant human leptin is a protein produced using recombinant DNA technology. It is structurally and functionally identical to the natural human leptin protein.
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Leptin and LEPR antibodies are laboratory reagents used in research applications. Leptin is a hormone involved in regulating energy balance, appetite, and metabolism. LEPR is the receptor for leptin. These antibodies can be used to detect and study the expression and distribution of leptin and its receptor in biological samples.
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Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.
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The Cell Counting Kit-8 (CCK-8) is a colorimetric assay used to measure the number of viable cells in cell proliferation and cytotoxicity assays. It utilizes a water-soluble tetrazolium salt that is reduced by cellular dehydrogenases, resulting in the formation of a colored formazan dye. The amount of formazan dye is directly proportional to the number of living cells in the culture, which can be quantified by measuring the absorbance of the solution.
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The Bicinchoninic acid (BCA) protein assay kit is a colorimetric detection and quantification method for determining the total protein concentration in a sample. The assay is based on the reduction of Cu2+ to Cu1+ by protein in an alkaline medium, and the subsequent chelation of the Cu1+ ion with bicinchoninic acid, resulting in a purple-colored reaction that absorbs light at 562 nm.
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Hoechst 33258 is a fluorescent dye that binds to the minor groove of DNA. It has excitation and emission wavelengths of 352 nm and 461 nm, respectively.
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Ab8227 is a mouse monoclonal antibody that recognizes the human CD19 protein. CD19 is a cell surface antigen expressed on B cells and some B cell malignancies. This antibody can be used for the detection and analysis of CD19-positive cells in various applications, such as flow cytometry and immunohistochemistry.
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Recombinant human leptin is a laboratory product that is a synthetic version of the human leptin protein. Leptin is a hormone produced by adipose tissue that regulates energy balance and metabolism. This recombinant leptin can be used for research purposes to study the biological functions of leptin.
More about "Leptin receptor, human"
The leptin receptor, also known as LEPR or OB-R, is a crucial cell surface receptor that binds to the hormone leptin.
Leptin is a key regulator of energy balance, food intake, and body weight.
The leptin receptor is expressed in various tissues, particularly the hypothalamus, where it mediates leptin's effects on appetite and metabolism.
Proper functioning of the leptin receptor is essential for maintaining normal body weight and energy homeostasis.
Dysfunctions in the leptin receptor pathway have been implicated in obesity and related metabolic disorders.
Recombinant human leptin, which mimics the structure and function of natural leptin, is a valuable tool for studying the leptin receptor and its signaling pathways.
Prism 6, a data analysis software, can be used to visualize and interpret the results of leptin receptor research.
Leptin and LEPR antibodies are also commonly used to detect and quantify the expression of the leptin receptor in different cell types and tissues.
Cell culture experiments investigating the leptin receptor often utilize fetal bovine serum (FBS) as a growth supplement.
The Cell Counting Kit-8 (CCK-8) is a convenient method for assessing cell viability and proliferation in these experiments.
Additionally, the Bicinchoninic acid (BCA) protein assay kit can be used to measure the total protein content in cell lysates, which is important for normalizing experimental data.
The Hoechst 33258 dye is a fluorescent stain that can be used to visualize and quantify the nuclei of cells, providing insights into cellular processes related to the leptin receptor.
By incorporating these related terms, abbreviations, and experimental techniques, researchers can optimize their leptin receptor studies for enhanced reproducibility and accuracy, as highlighted by the Metadescription of PubCompare.ai.
This comprehensive understanding of the leptin receptor and its associated tools and methodologies can lead to more reliable and informative results in the pursuit of understanding obesity and related metabolic disorders.
Leptin is a key regulator of energy balance, food intake, and body weight.
The leptin receptor is expressed in various tissues, particularly the hypothalamus, where it mediates leptin's effects on appetite and metabolism.
Proper functioning of the leptin receptor is essential for maintaining normal body weight and energy homeostasis.
Dysfunctions in the leptin receptor pathway have been implicated in obesity and related metabolic disorders.
Recombinant human leptin, which mimics the structure and function of natural leptin, is a valuable tool for studying the leptin receptor and its signaling pathways.
Prism 6, a data analysis software, can be used to visualize and interpret the results of leptin receptor research.
Leptin and LEPR antibodies are also commonly used to detect and quantify the expression of the leptin receptor in different cell types and tissues.
Cell culture experiments investigating the leptin receptor often utilize fetal bovine serum (FBS) as a growth supplement.
The Cell Counting Kit-8 (CCK-8) is a convenient method for assessing cell viability and proliferation in these experiments.
Additionally, the Bicinchoninic acid (BCA) protein assay kit can be used to measure the total protein content in cell lysates, which is important for normalizing experimental data.
The Hoechst 33258 dye is a fluorescent stain that can be used to visualize and quantify the nuclei of cells, providing insights into cellular processes related to the leptin receptor.
By incorporating these related terms, abbreviations, and experimental techniques, researchers can optimize their leptin receptor studies for enhanced reproducibility and accuracy, as highlighted by the Metadescription of PubCompare.ai.
This comprehensive understanding of the leptin receptor and its associated tools and methodologies can lead to more reliable and informative results in the pursuit of understanding obesity and related metabolic disorders.