To facilitate cross-species comparison of methylomes of multiple species, we converted the methylation data of mouse and chimpanzee to the corresponding locations in the human genome. The liftOver tool provided by UCSC Genome Browser http://genome.ucsc.edu/cgi-bin/hgLiftOver is used to directly convert the methylation level file at individual cytosines (output from the program methcounts ) to the human genome. Next we rerun the hmr and pmd programs on the converted methylation data file. Since the program amr works on the mapped read file, we directly liftover the list of AMRs with the liftOver program.
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Amino Acid
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AREG protein, human
AREG protein, human
AREG (Amphiregulin) is a member of the epidermal growth factor (EGF) family of proteins that plays a key role in cell proliferation, differentiation, and survival.
It is involved in various physiological and pathological processes, including tissue development, wound healing, and cancer progression.
AREG binds to and activates the EGF receptor (EGFR), triggering downstream signaling cascades that regulate gene expression and cellular behavior.
Researchers studying AREG protein and its functions can utilize PubCompare.ai, a powerful tool that helps identify the most effective protocols and procedures from literature, preprints, and patents.
This AI-driven comparison platform can enhance the reproducibility and accuracy of AREG protein experiments, allowing researchers to discover the best products and methodologies to advance their studies.
It is involved in various physiological and pathological processes, including tissue development, wound healing, and cancer progression.
AREG binds to and activates the EGF receptor (EGFR), triggering downstream signaling cascades that regulate gene expression and cellular behavior.
Researchers studying AREG protein and its functions can utilize PubCompare.ai, a powerful tool that helps identify the most effective protocols and procedures from literature, preprints, and patents.
This AI-driven comparison platform can enhance the reproducibility and accuracy of AREG protein experiments, allowing researchers to discover the best products and methodologies to advance their studies.
Most cited protocols related to «AREG protein, human»
AREG protein, human
Cytosine
Epigenome
Genome
Genome, Human
Methylation
Mice, House
Pan troglodytes
Species Specificity
Amino Acids, Essential
AREG protein, human
beta-D-Galactoside alpha 2-6-Sialyltransferase
Bos taurus
Cell Lines
Cells
Cloning Vectors
Dietary Supplements
epidermal growth factor receptor VIII
Fetal Bovine Serum
Gentamicin
Glucose
Glutamine
Insulin
Madin Darby Canine Kidney Cells
Parent
Penicillins
Puromycin
Serum
Short Hairpin RNA
Streptomycin
AREG protein, human
Childbirth
Climate
Climate Change
Dietary Supplements
Genetic Heterogeneity
Head
Head of Household
Households
Migrants
Motivation
Negroid Races
Range of Motion, Articular
To identify allele-specific methylated regions, we use the linkage information of the methylation status between adjacent cytosines in a read. The separation of reads into two alleles and the testing of whether a certain region fits the allele-specific model is carried out with the statistical method described by Fang et al. [17] . Additionally, a single-site profile for an allele-specific methylation “score” can be computed along the genome by testing for significance of linkage between methylation status in reads covering adjacent CpGs. The programs for identifying AMRs and computing allelic scores are amrfinder and allelicmeth.
Alleles
AREG protein, human
cytidylyl-3'-5'-guanosine
Cytosine
Genome
Methylation
Seizures
AREG protein, human
Biological Assay
Deoxyribonuclease I
Digestion
DNA, Complementary
EREG protein, human
Freezing
Gene Expression
Heparin-binding EGF-like Growth-Factor
Reverse Transcription
ribosomal protein P0
RPLP0 protein, human
Skin
TGFA protein, human
Most recents protocols related to «AREG protein, human»
Both cell lysates and EV preparations, which were lysed in RIPA or 1X sample buffer, were electrophoretically separated using 10% mini‐PROTEAN precast gels (BioRad). Gels were loaded for western analysis with EV lysates extracted from the same protein mass of secreting cells, so that changes in band intensity on the blots with glutamine depletion reflected a net change in secretion of the marker on a per cell basis (see Fan et al., 2020 (link)). Protein preparations were ultimately dissolved in either reducing (containing 5% β‐mercaptoethanol) or non‐reducing (for CD63 and CD81 detection) sample buffer and were heated to 90°C–100°C for 10 min before loading with a pre‐stained protein ladder (Bio‐Rad). Proteins were wet‐transferred to polyvinylidene difluoride (PVDF) membranes at 100 V for 1 h using a Mini Trans‐Blot Cell (Bio‐Rad). Membranes were then blocked with either 5% milk (CD63 detection) or 5% BSA in TBS buffer with Tween‐20 (TBST) for 30 min and probed overnight at 4°C with primary antibody diluted in blocking buffer. The membranes were washed for 3 × 10 min with TBST, then probed with the relevant secondary antibodies for 1 h at 22°C, washed for 3 × 10 min again, and then the signals detected using the enhanced chemiluminescent detection reagent (Clarity, BioRad) and the Touch Imaging System (BioRad). Relative band intensities were quantified by ChemiDoc software (Bio‐Rad) or ImageJ. Signals were normalised to cell lysate protein (Fan et al., 2020 (link)).
Antibody suppliers, catalogue numbers and concentrations used were: rabbit anti‐CHMP1a (Proteintech #15761‐1‐AP, 1:500), rabbit anti‐CHMP1b (Proteintech #14639‐1‐AP, 1:500), rabbit anti‐IST1 (Biorad #VPA00314, 1:500), mouse anti‐CHMP5 (Santa Cruz #sc‐374338, 1:500), rabbit anti‐4E‐BP1 (Cell Signaling Technology #9644, rabbit anti‐p‐4E‐BP1‐Ser65 (Cell Signaling Technology #9456, 1:1000), rabbit anti‐S6 (Cell Signaling Technology #2217, 1:4000), rabbit anti‐p‐S6‐Ser240/244 S6 (Cell Signalling Technology #5364, 1:4000), rabbit anti‐Caveolin‐1 (Cell Signaling Technology #3238, 1:500), goat anti‐AREG (R&D Systems #AF262, 1:200), mouse anti‐Tubulin (Sigma #T8328, 1:4000), mouse anti‐CD81 (Santa Cruz #23962, 1:500), mouse anti‐CD63 (BD Biosciences # 556019, 1:500), rabbit anti‐Syntenin‐1 antibody (Abcam ab133267, 1:500), rabbit anti‐Tsg101 (Abcam ab125011, 1:500), mouse anti‐Rab11 (BD Biosciences #610657, 1:500), sheep anti‐TGN46 (BioRad; AHP500G, 1:1000), rabbit anti‐EEA1 (Cell Signalling Technology #3288, 1:1000) , anti‐mouse IgG (H+L) HRP conjugate (Promega #W4021, 1:10000), anti‐rabbit IgG (H+L) HRP conjugate (Promega #W4011, 1:10000), anti‐goat IgG (H+L) HRP conjugate (R&D Systems #HAF109, 1:100).
Antibody suppliers, catalogue numbers and concentrations used were: rabbit anti‐CHMP1a (Proteintech #15761‐1‐AP, 1:500), rabbit anti‐CHMP1b (Proteintech #14639‐1‐AP, 1:500), rabbit anti‐IST1 (Biorad #VPA00314, 1:500), mouse anti‐CHMP5 (Santa Cruz #sc‐374338, 1:500), rabbit anti‐4E‐BP1 (Cell Signaling Technology #9644, rabbit anti‐p‐4E‐BP1‐Ser65 (Cell Signaling Technology #9456, 1:1000), rabbit anti‐S6 (Cell Signaling Technology #2217, 1:4000), rabbit anti‐p‐S6‐Ser240/244 S6 (Cell Signalling Technology #5364, 1:4000), rabbit anti‐Caveolin‐1 (Cell Signaling Technology #3238, 1:500), goat anti‐AREG (R&D Systems #AF262, 1:200), mouse anti‐Tubulin (Sigma #T8328, 1:4000), mouse anti‐CD81 (Santa Cruz #23962, 1:500), mouse anti‐CD63 (BD Biosciences # 556019, 1:500), rabbit anti‐Syntenin‐1 antibody (Abcam ab133267, 1:500), rabbit anti‐Tsg101 (Abcam ab125011, 1:500), mouse anti‐Rab11 (BD Biosciences #610657, 1:500), sheep anti‐TGN46 (BioRad; AHP500G, 1:1000), rabbit anti‐EEA1 (Cell Signalling Technology #3288, 1:1000) , anti‐mouse IgG (H+L) HRP conjugate (Promega #W4021, 1:10000), anti‐rabbit IgG (H+L) HRP conjugate (Promega #W4011, 1:10000), anti‐goat IgG (H+L) HRP conjugate (R&D Systems #HAF109, 1:100).
2-Mercaptoethanol
anti-IgG
Antibodies
Antibodies, Anti-Idiotypic
AREG protein, human
Buffers
Caveolin 1
Cells
Domestic Sheep
EIF4EBP1 protein, human
Gels
Glutamine
Goat
Immunoglobulins
Milk, Cow's
Mus
polyvinylidene fluoride
Promega
Proteins
Proto-Oncogene Mas
Rabbits
Radioimmunoprecipitation Assay
secretion
Syntenin-1
Tissue, Membrane
Touch
TSG101 protein, human
Tubulin
Tween 20
Western blot was performed as previously described [71 (link)]. In brief, total proteins of cultured cells were lysed in lysis buffer (150 mM NaCl, 50 mM Tris- HCl, pH 7.5, 5 mM EDTA, 1% Triton X-100, 10 mM NaF, 1 mM Na3VO3, 0.1% SDS, and 1% Sodium deoxycholate) supplemented with protease inhibitor cocktail (4693116001, Roche, Basel, Switzerland). The supernatants collected from cultured epithelial Ishikawa cells with serum-free culture medium were dialyzed in 2 L distilled water for 2 h at room temperature with stirring and lyophilized in a vacuum lyophilizer (FreeZone Plus, LABCONCO, Kansas, MO, USA) for 8 h. The lyophilized powder was dissolved in lysis buffer. The BCA kit (23225, Thermo Scientific, Shanghai, China) were used to measure protein concentration. Protein lysates (5–10 μg) were electrophoresed in SDS-PAGE gels and transferred onto polyvinylidene fluoride (PVDF) membranes (IPVH00010, Millipore, Billerica, MA, USA). Membranes were blocked with 5% non-fat milk (A600669, Sangon, Shanghai, China) for 1 h at room temperature and then incubated with the specific primary antibody overnight at 4 °C. After the membranes were incubated with horseradish peroxidase (HRP)-conjugated secondary antibody (1:5000, Invitrogen, Carlsbad, CA, USA) for 1 h at room temperature, the signals were detected with an ECL Chemiluminescent Kit (WBKLS0100, Millipore, Burlington, MA, USA) on Tanon Imaging System (5200, Tanon, Shanghai, China). The primary antibodies used in this study included anti-Tubulin (#2144, 1:1000, Cell Signaling, Danvers, MA, USA), anti-GAPDH (#sc-32233, 1:1000, Santa Cruz, CA, USA), anti-ODC1 (#28728-1-AP, 1:1000, Proteintech, Rosemont, IL, USA), anti-IL4I1 (#ab222102, 1:1000, Abcam, Cambridge, UK), anti-Integrin β3 (#13166s, 1:1000, Cell Signaling Technology), anti-HOXA10 (#Sc-28620, 1:1000, Santa Cruz), anti-FOXO1 (#2880, 1:1000, Cell Signaling Technology), anti-AREG (#A1860, 1:500, Abclonal, Woburn, MA, USA), anti-Lamin A/C (#2032s, 1:1000, Cell Signaling Technology), anti-AHR (#NB100-2289SS, 1:1000, Novus Biologicals, Englewood, CO, USA).
Antibodies
AREG protein, human
Biological Factors
Buffers
Cultured Cells
Culture Media
Deoxycholic Acid, Monosodium Salt
Edetic Acid
Epithelial Cells
GAPDH protein, human
Gels
Horseradish Peroxidase
Immunoglobulins
Integrins
LMNA protein, human
Milk, Cow's
Novus
polyvinylidene fluoride
Powder
Protease Inhibitors
Proteins
SDS-PAGE
Serum
Sodium Chloride
Tissue, Membrane
Triton X-100
Tromethamine
Tubulin
Vacuum
Western Blotting
RIPA buffer containing 50 mM tris (pH 7.4), 150 mM NaCl, 1% NP-40, 0.5% sodium deoxycholate and 0.1% SDS was used for cell lysis (Beyotime). Then, 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was to achieve protein separation. Polyvinylidene difluoride (PVDF) membrane (Millipore) was used for protein transfer. After blocking with a 5% blocking solution, the PVDF membrane was incubated with the primary antibody overnight at 4 °C. Following incubation with appropriate secondary antibodies, treatment with Luminata Crescendo Western HRP substrate (Millipore) was performed as well as exposure and digital imaging. The primary antibodies of TPM2 (1:1000, Proteintech, 28587-1-AP), PDLIM7 (1:1000, Proteintech, 10221-1-AP), YAP1 (1:1000, Proteintech, 13584-1-AP), p-YAP1(S127) (1:1000, Abcam, ab76252), NKX3.1 (1:1000, Abcam, ab196020), PSA (1:1000, Abcam, ab76113), c-Myc (1:1000, Cell Signaling Technology, 18583), AREG (1:1000, Proteintech, 16036-1-AP), PCAN (1:1000, Proteintech, 10205-2-AP), HA tag polyclonal antibody (1:1000, Proteintech, 51064-2-AP), GST tag polyclonal antibody (1:1000, Proteintech, 10000-0-AP), MYC tag polyclonal antibody (1:1000, Proteintech, 16286-1-AP) and GAPDH (1:2000, Proteintech, 10494-1-AP). The Beyotime cytoplasmic protein extraction kit was used to extract nuclear proteins based on the manufacturer’s instructions.
Antibodies
AREG protein, human
Buffers
Cells
Cytoplasm
Deoxycholic Acid, Monosodium Salt
Fingers
GAPDH protein, human
Immunoglobulins
Nonidet P-40
Nuclear Proteins
Oncogenes, myc
polyvinylidene fluoride
Proteins
Radioimmunoprecipitation Assay
SDS-PAGE
Sodium Chloride
Tissue, Membrane
Tromethamine
YAP1 protein, human
Human UM cell line (MUM-2B) was obtained from the Fuheng Biology Inc., (Fuheng, Shanghai, China). For MUM-2B cell culture, DMEM (keyGEN bioTECH, China) with 10% fetal bovine serum (FBS) was used. Cells were transfected with the synthesized siRNAs (GenePharma, China) targeting AREG by the Lipofectamine3000 based on the manufacturer’s protocol. The siRNA-AREG sequences are provided in Supplementary Table S2 .
AREG protein, human
Cell Culture Techniques
Cell Lines
Cells
Fetal Bovine Serum
Homo sapiens
RNA, Small Interfering
The cell total RNA of was collected using RNA easy reagent (Vazyme, China) and cDNA was obtained using a PrimeScript RT Reagent Kit (Takara, Japan). Then, qRT-PCR was performed through a ChamQ SYBR qPCR Master Mix (Vazyme, China). The relative expression levels of m RNA were normalized to GAPDH. The primer sequences of AREG and GAPDH are shown in Supplementary Table S1 .
AREG protein, human
DNA, Complementary
GAPDH protein, human
Oligonucleotide Primers
Transcription, Genetic
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The RNeasy Mini Kit is a laboratory equipment designed for the purification of total RNA from a variety of sample types, including animal cells, tissues, and other biological materials. The kit utilizes a silica-based membrane technology to selectively bind and isolate RNA molecules, allowing for efficient extraction and recovery of high-quality RNA.
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TRIzol reagent is a monophasic solution of phenol, guanidine isothiocyanate, and other proprietary components designed for the isolation of total RNA, DNA, and proteins from a variety of biological samples. The reagent maintains the integrity of the RNA while disrupting cells and dissolving cell components.
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The High-Capacity cDNA Reverse Transcription Kit is a laboratory tool used to convert RNA into complementary DNA (cDNA) molecules. It provides a reliable and efficient method for performing reverse transcription, a fundamental step in various molecular biology applications.
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TRIzol is a monophasic solution of phenol and guanidine isothiocyanate that is used for the isolation of total RNA from various biological samples. It is a reagent designed to facilitate the disruption of cells and the subsequent isolation of RNA.
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The IScript cDNA Synthesis Kit is a reagent kit used for the reverse transcription of RNA into complementary DNA (cDNA). The kit contains all the necessary components to perform this reaction, including a reverse transcriptase enzyme, reaction buffer, and oligo(dT) primers.
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The StepOnePlus Real-Time PCR System is a compact, flexible, and easy-to-use instrument designed for real-time PCR analysis. It can be used to detect and quantify nucleic acid sequences.
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The RNeasy kit is a laboratory equipment product that is designed for the extraction and purification of ribonucleic acid (RNA) from various biological samples. It utilizes a silica-membrane-based technology to efficiently capture and isolate RNA molecules.
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TaqMan Gene Expression Assays are a set of pre-designed and pre-optimized qPCR assays for accurately quantifying gene expression levels. They provide a sensitive and reliable method for measuring targeted mRNA transcripts in a variety of sample types.
Sourced in United Kingdom
Recombinant AREG is a protein produced in a laboratory using recombinant DNA technology. It is the human Amphiregulin (AREG) protein.
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Lipofectamine 2000 is a cationic lipid-based transfection reagent designed for efficient and reliable delivery of nucleic acids, such as plasmid DNA and small interfering RNA (siRNA), into a wide range of eukaryotic cell types. It facilitates the formation of complexes between the nucleic acid and the lipid components, which can then be introduced into cells to enable gene expression or gene silencing studies.
More about "AREG protein, human"
Amphiregulin (AREG) is a member of the epidermal growth factor (EGF) family of proteins, playing a crucial role in cell proliferation, differentiation, and survival.
It is involved in various physiological and pathological processes, such as tissue development, wound healing, and cancer progression.
AREG binds to and activates the EGF receptor (EGFR), triggering downstream signaling cascades that regulate gene expression and cellular behavior.
Researchers studying AREG protein and its functions can utilize PubCompare.ai, a powerful AI-driven comparison platform that helps identify the most effective protocols and procedures from literature, preprints, and patents.
This tool can enhance the reproducibility and accuracy of AREG protein experiments, allowing researchers to discover the best products and methodologies to advance their studies.
To extract and analyze AREG RNA and protein expression, researchers may employ techniques like the RNeasy Mini Kit, TRIzol reagent, and High-Capacity cDNA Reverse Transcription Kit.
These tools enable efficient RNA extraction, cDNA synthesis, and real-time PCR analysis using the StepOnePlus Real-Time PCR System and TaqMan Gene Expression Assays.
Moreover, researchers can investigate the functional aspects of AREG by working with recombinant AREG protein and utilizing transfection methods like Lipofectamine 2000 to introduce AREG-encoding plasmids into cell lines.
These approaches can help elucidate the signaling pathways and biological processes regulated by AREG.
In summary, the AREG protein is a versatile and influential member of the EGF family, with significant implications in various physiological and pathological contexts.
By leveraging PubCompare.ai and employing established molecular biology techniques, researchers can optimize their AREG protein research, enhance reproducibility, and drive their studies forward with greater efficiency and accuracy.
It is involved in various physiological and pathological processes, such as tissue development, wound healing, and cancer progression.
AREG binds to and activates the EGF receptor (EGFR), triggering downstream signaling cascades that regulate gene expression and cellular behavior.
Researchers studying AREG protein and its functions can utilize PubCompare.ai, a powerful AI-driven comparison platform that helps identify the most effective protocols and procedures from literature, preprints, and patents.
This tool can enhance the reproducibility and accuracy of AREG protein experiments, allowing researchers to discover the best products and methodologies to advance their studies.
To extract and analyze AREG RNA and protein expression, researchers may employ techniques like the RNeasy Mini Kit, TRIzol reagent, and High-Capacity cDNA Reverse Transcription Kit.
These tools enable efficient RNA extraction, cDNA synthesis, and real-time PCR analysis using the StepOnePlus Real-Time PCR System and TaqMan Gene Expression Assays.
Moreover, researchers can investigate the functional aspects of AREG by working with recombinant AREG protein and utilizing transfection methods like Lipofectamine 2000 to introduce AREG-encoding plasmids into cell lines.
These approaches can help elucidate the signaling pathways and biological processes regulated by AREG.
In summary, the AREG protein is a versatile and influential member of the EGF family, with significant implications in various physiological and pathological contexts.
By leveraging PubCompare.ai and employing established molecular biology techniques, researchers can optimize their AREG protein research, enhance reproducibility, and drive their studies forward with greater efficiency and accuracy.