All experiments were performed with HeLa cells44 (link) (obtained from E.V. Makayev, Nanyang Technological University, Singapore) either stably or transiently transfected with a plasmid for bicistronic expression of mCherry–eDHFR and indicated Haloenzyme constructs (see below). CENPB and mitochondria experiments were performed with stable cell lines; centrosome and kinetochore experiments were performed by transient transfection. Stable cell lines were created by recombinase-mediated cassette exchange using the HILO recombinase-mediated cassette exchange system44 (link) (obtained from E.V. Makeyev, Nanyang Technological University, Singapore). Briefly: a monoclonal acceptor cell line with LoxP and Lox2272 recombination sites at a single chromosomal locus was cotransfected with a donor plasmid containing a transgenic cassette flanked by LoxP and Lox2272 sites and a second plasmid expressing Cre recombinase, followed by selection for a marker in the donor cassette. Cells were cultured in growth medium (Dulbecco’s Modified Eagle’s medium with 10% FBS and penicillin–streptomycin) at 37 °C in a humidified atmosphere with 5% CO2. Cells at ~ 60% confluency in a single well of a 6-well plate were transfected with 1 µg of donor plasmid + 10 ng of Cre plasmid pEM784 (see below for plasmid details) using 3 µl of Fugene 6 (Promega). Two days after transfection, 1 µg ml−1 puromycin was added to the growth medium for the selection of stable cell lines. Transient transfections were performed as above using 3 µl of Fugene 6 and 1 µg of plasmid DNA, 40 h before experiment.
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CENPB protein, human
CENPB protein, human
CENPB (Centromere Protein B) is a crucial component of the human centromere, playing a vital role in chromosome segregation and cell division.
This protein binds to specific DNA sequences within the centromeric region, contributing to the structural organization and function of the centromere.
CENPB has been implicated in various cellular processes, including chromosome stability, kinetochore assembly, and the regulation of gene expression.
Understanding the role of CENPB in human biology can provide important insights into the mechanisms underlying chromosomal abnormalities and related diseases.
Reseachers can leverage PubCompare.ai's AI-powered platform to discover the latest insights on CENPB, locate optimized research protocols, and identify the best products for their studies, ultimately boosting reproducibility and streamlining their research effrots.
This protein binds to specific DNA sequences within the centromeric region, contributing to the structural organization and function of the centromere.
CENPB has been implicated in various cellular processes, including chromosome stability, kinetochore assembly, and the regulation of gene expression.
Understanding the role of CENPB in human biology can provide important insights into the mechanisms underlying chromosomal abnormalities and related diseases.
Reseachers can leverage PubCompare.ai's AI-powered platform to discover the latest insights on CENPB, locate optimized research protocols, and identify the best products for their studies, ultimately boosting reproducibility and streamlining their research effrots.
Most cited protocols related to «CENPB protein, human»
Animals, Transgenic
Atmosphere
Cell Lines
Cells
CENPB protein, human
Centrosome
Chromosomes
Cre recombinase
Culture Media
Eagle
FuGene
HeLa Cells
Kinetochores
Mitochondria
Penicillins
Plasmids
Promega
Puromycin
Recombinase
Recombination, Genetic
Streptomycin
Tissue Donors
Transfection
Transients
The complete rat MeCP2 ORF as well as the DNA-binding domain of human CENPB (aa 1–169; Shelby et al., 1996 (link)) were fused in frame at the NH2 terminus of the enhanced YFP (pEYFP-N1 vector; CLONTECH Laboratories, Inc.). The YFP-containing part from MeCP2-YFP fusion was cut out and replaced by either enhanced GFP (isolated from pEGFP-N1 vector; CLONTECH Laboratories, Inc.) or mRFP1 (Campbell et al., 2002 (link)) to construct MeCP2-GFP and MeCP2-mRFP1, respectively. MeCP2 deletion mutants were generated from the above plasmids using conveniently located restriction enzyme sites or by PCR amplification using primers including compatible restriction sites (primer sequences are available upon request). The complete human DNA ligase I ORF was fused in frame at the COOH terminus of the DsRed1 gene (pDsRed1-C1; CLONTECH Laboratories, Inc.) (Easwaran et al., 2004 (link), 2005 (link)). The GFP-HP1α (human HP1α; Cheutin et al., 2003 (link)); GFP-MBD2 (human MBD2a; Tatematsu et al., 2000 (link)); MBD1-GFP, MBD4-GFP, and GFP-MBD3 (mouse cDNAs for MBD1, MBD4, and MBD3; Hendrich and Bird, 1998 (link)); and FLAG-HP1β (FLAG epitope–tagged mouse HP1β; Nielsen et al., 2001 (link)) were as described. Correct expression of all fusions in mammalian cells was checked by Western blot analysis as described before (Cardoso et al., 1997 (link); Easwaran et al., 2005 (link)).
Aves
CBX5 protein, human
Cells
CENPB protein, human
Cloning Vectors
Deletion Mutation
DNA, Complementary
DNA Restriction Enzymes
Epitopes
Genes
Homo sapiens
Ligase I, DNA
Mammals
MBD1 protein, human
MBD2 protein, human
MBD4 protein, human
MECP2 protein, human
methyl-CpG binding domain protein 3, human
Mice, Laboratory
Oligonucleotide Primers
Plasmids
Reading Frames
Western Blot
Cells
CENPB protein, human
Clone Cells
Genes
Homo sapiens
KIF18A protein, human
Kinesin
Plasmids
Reading Frames
RNA, Small Interfering
Trimethoprim-Sulfamethoxazole Combination
Detailed methods are described in online supplementary file 1 . We created 256 recombinant antibodies of ASCs in salivary glands from nine patients. The reactivity of these antibodies were investigated by ELISA and a newly developed procedure, antigen-binding beads assay. Antibodies against CBX5, CENP-A, CENP-B, CENP-C and MIS12 complex (MIS12C) were examined in 269 serum and 11 salivary gland tissues.
Antibodies
Antigens
Biological Assay
CENPB protein, human
centromere protein C
Enzyme-Linked Immunosorbent Assay
Patients
Salivary Glands
Serum
STS protein, human
Tissues
Cells were fixed for 15 min in PBS-buffered 4% paraformaldehyde, followed by permeablization for 5 min in Triton-X buffer68 (link). The following antibodies were used at a concentration of 1–2 μg ml−1 for immunofluorescence assays: anti-CENP-B (sc22788; Santa Cruz Biotechnology), anti-BubR1 (612503), anti-α-tubulin (T9026; Sigma-Aldrich), anti-p53 (sc126; Santa Cruz Biotechnology), anti-53BP1 (sc22760; Santa Cruz Biotechnology), anti-p62 (sc28359; Santa Cruz Biotechnology) and anti-LC3B (3868; Cell Signaling Technology). Images were captured with a Plan-APOCHROMAT × 100 oil lens under an Axiovert 200M microscope (Carl Zeiss).
alpha-Tubulin
Antibodies
Cells
CENPB protein, human
Fluorescent Antibody Technique, Direct
Lens, Crystalline
Microscopy
paraform
TP53BP1 protein, human
Most recents protocols related to «CENPB protein, human»
To select sequences for the first N-zip library, only genes with significant enrichment (adjusted P < 0.05) in the analyzed primary neuronal datasets were considered as candidates to generate a list of genes with reliable neurite localization. This selection was further restricted to genes for which an enrichment value could be calculated in our PCN system (log2FC not NA). Then, genes with (1) a significant enrichment in at least four datasets; (2) median log2FC > 1; and (3) either mean log2FC > 1 or a positive log2FC value in all datasets were chosen. Additionally, genes with a significant enrichment value in at least five datasets and either median log2FC > 1 or mean log2FC > 1 or a positive log2FC value in all datasets were chosen as well.
This initial unbiased set of genes was then manually refined by (1) excluding genes encoded by the mitochondrial genome as well as some genes with the annotated nuclear or mitochondrial function (Pola1, Ezh2, Smc4, Cenpb, Pink1 and Ncl); (2) adding genes with a known zipcode or neurite localization sequence (Camk2a94 (link), Actb5 (link), Bdnf9 (link), Arc11 (link),95 (link), Cdc42 (ref. 25 (link)), Map2 (ref. 96 (link)) and Bc1 (ref. 97 (link)); (3) adding genes that showed localization in non-primary31 (link) and in-house datasets as well as our PCN and fewer other primary datasets (Rab13, Net1, Hmgn5, 2410006H16Rik, Pfdn5, Tagln2, Pfdn1 and Cryab); and (4) restricting the genes encoding for ribosomal proteins and translation factors to a smaller subset with sufficiently large 3′ UTRs (Rplp2, Rpl12, Rpl39, Rpl37, Rpl14, Rps28, Rpsa, Rps24, Rps23, Rps18, Eef1b2, Eef1a1 and Eef1g).
This initial unbiased set of genes was then manually refined by (1) excluding genes encoded by the mitochondrial genome as well as some genes with the annotated nuclear or mitochondrial function (Pola1, Ezh2, Smc4, Cenpb, Pink1 and Ncl); (2) adding genes with a known zipcode or neurite localization sequence (Camk2a94 (link), Actb5 (link), Bdnf9 (link), Arc11 (link),95 (link), Cdc42 (ref. 25 (link)), Map2 (ref. 96 (link)) and Bc1 (ref. 97 (link)); (3) adding genes that showed localization in non-primary31 (link) and in-house datasets as well as our PCN and fewer other primary datasets (Rab13, Net1, Hmgn5, 2410006H16Rik, Pfdn5, Tagln2, Pfdn1 and Cryab); and (4) restricting the genes encoding for ribosomal proteins and translation factors to a smaller subset with sufficiently large 3′ UTRs (Rplp2, Rpl12, Rpl39, Rpl37, Rpl14, Rps28, Rpsa, Rps24, Rps23, Rps18, Eef1b2, Eef1a1 and Eef1g).
CDC42 protein, human
CENPB protein, human
DNA Library
EEF1A1 protein, human
EZH2 protein, human
factor A
Genes
Genes, vif
Genome, Mitochondrial
Lamr1 protein, human
METAP2 protein, human
Mitochondrial Inheritance
Neurites
Neurons
Ribosomal Proteins
Untranslated Regions
The correlation between ceRNA networks was analyzed by IBM SPSS software Version 26.0 (SPSS Inc., Chicago, IL, USA) and GraphPad Prism 9.0.0 (GraphPad, La Jolla, CA, USA). Correlations between ceRNA networks and clinical data, such as high-resolution CT (HRCT), antinuclear antibody profile (Scl-70, CENP-B, Ro-52), C-reactive protein (CRP), IL-10, lymphocyte percentage, and neutrophil percentage, were analyzed, and then the receiver operating characteristic curve (ROC) for SSc diagnosis was drawn.
Antibodies, Antinuclear
CENPB protein, human
C Reactive Protein
Diagnosis
IL10 protein, human
Lymphocyte
Neutrophil
prisma
Cells were cultured in growth medium and 50 ng/mL Nocodazole were added 16 h prior to fixation. Metaphase spreads were prepared as previously described. Telomere and centromeres were concomitantly hybridized with TelC-Cy3 (PNA Bio) and CENPB-Cy5 (PNA Bio) for 2 h at RT after denaturation at 75 °C. After dehydration, slides were stained with 1 µg/mL DAPI (KPL) and mounted with ProLong Gold before imaging.
Cells
CENPB protein, human
Centromere
Culture Media
DAPI
Dehydration
Gold
Metaphase
Nocodazole
Telomere
The study was cross-sectional and prospective in design. It was carried out between 2012 and 2020 within the population of outpatients and inpatients of the Department of Rheumatology and Internal Medicine in Poznan, Poland. The study population consisted of 96 adult patients with SSc, classified using the American College of Rheumatology and the European League Against Rheumatism (ACR-EULAR) 2013 criteria [12 (link)]. There were no exclusion criteria.
History taking and physical examination of all patients were carried out by the same physician (P.Ż.), who has experience in the care of SSc patients. Clinical data were collected using MEDS (Minimal Essential Data Set) in line with the EUSTAR (EULAR Scleroderma Trials and Research group) methodology [13 (link)], also by the same practitioner. The following features were assessed: gender, age, time of disease since the onset of Raynaud’s phenomenon, time of disease since the onset of other symptoms characteristic of SSc, duration of the disease since the diagnosis, skin involvement using the modified Rodnan skin score (mRSS), Raynaud’s phenomenon, digital ulcerations, synovitis, joint contractures, muscle weakness, esophageal symptoms (heartburn and dysphagia), intestinal manifestations (diarrhea, constipation, and bloating), arterial hypertension, scleroderma renal crisis (ever), dyspnea, and palpitations.
Specialist cardiac (echocardiography) and pulmonary examination (pulmonary function tests and high-resolution computed tomography) were carried out. Certified specialist physicians interpreted the results: a cardiologist (T.M.-K.) and a pulmonologist (T.P.), both with considerable experience in SSc diagnostics. A number of characteristics were sought: heart conduction blocks; palpitations, systolic and diastolic dysfunction; ejection fraction; pulmonary arterial hypertension using Doppler ultrasonography; total lung capacity; pulmonary restriction using pulmonary function tests; and the presence of SSc-related interstitial lung disease (ILD) using HRCT.
Sera of patients were secured and assessed for autoantibodies. An immunoblot test known as Systemic Sclerosis Profile (SSP) by EUROIMMUN Medizinische Labordiagnostika AG (Lübeck, Germany) was used to assess SSc-related autoantibodies (Scl-70, centromere protein A, centromere protein B, RNA Pol III 11 kDa, RNA Pol III 155 kDa, fibrillarin, Nor90, Th/To, PM-Scl100, PM-Scl75, Ku, PDGFR, and Ro-52) according to the manufacturer’s recommendations, including the cutoff for seropositivity. Similarly, we used an ANA profile immunoblot by the same manufacturer to detect Scl-70, CENP-B, and PM-Scl. In addition, we determined the erythrocyte sedimentation rate (ESR), C-reactive protein concentration, serum creatinine concentration, the estimated glomerular filtration rate (chronic kidney disease epidemiology collaboration equation; eGFR CKD), as well as the presence or absence of proteinuria.
Statistical analyses were conducted in Statistica 13.3 (TIBCO, Palo Alto, CA, USA) and Microsoft Excel 2016 (Redmond, WA, USA). The associations between autoantibody seropositivity and clinical manifestations or disease subset were assessed using Fisher’s exact test, whereas the comparison of the sensitivity of both immunoblots was made using McNemar’s chi-square test. The significance threshold was set at 0.05. In cases where the contingency table included at least one zero while calculating odds ratios, we used Haldane–Anscombe correction by adding 0.5 to all values in a table so that calculating OR was mathematically possible. Heatmap was created in R 4.1.3 (R Software Foundation, Vienna, Austria) using the pheatmap package 1.0.12, with default hierarchical clustering, after logarithmic transformation (base 10) of odds ratios.
This study obtained the approval of the Bioethics Committee at the Poznan University of Medical Sciences (No. 988/11). All patients participating in the study gave informed consent in writing in line with local law regulations.
History taking and physical examination of all patients were carried out by the same physician (P.Ż.), who has experience in the care of SSc patients. Clinical data were collected using MEDS (Minimal Essential Data Set) in line with the EUSTAR (EULAR Scleroderma Trials and Research group) methodology [13 (link)], also by the same practitioner. The following features were assessed: gender, age, time of disease since the onset of Raynaud’s phenomenon, time of disease since the onset of other symptoms characteristic of SSc, duration of the disease since the diagnosis, skin involvement using the modified Rodnan skin score (mRSS), Raynaud’s phenomenon, digital ulcerations, synovitis, joint contractures, muscle weakness, esophageal symptoms (heartburn and dysphagia), intestinal manifestations (diarrhea, constipation, and bloating), arterial hypertension, scleroderma renal crisis (ever), dyspnea, and palpitations.
Specialist cardiac (echocardiography) and pulmonary examination (pulmonary function tests and high-resolution computed tomography) were carried out. Certified specialist physicians interpreted the results: a cardiologist (T.M.-K.) and a pulmonologist (T.P.), both with considerable experience in SSc diagnostics. A number of characteristics were sought: heart conduction blocks; palpitations, systolic and diastolic dysfunction; ejection fraction; pulmonary arterial hypertension using Doppler ultrasonography; total lung capacity; pulmonary restriction using pulmonary function tests; and the presence of SSc-related interstitial lung disease (ILD) using HRCT.
Sera of patients were secured and assessed for autoantibodies. An immunoblot test known as Systemic Sclerosis Profile (SSP) by EUROIMMUN Medizinische Labordiagnostika AG (Lübeck, Germany) was used to assess SSc-related autoantibodies (Scl-70, centromere protein A, centromere protein B, RNA Pol III 11 kDa, RNA Pol III 155 kDa, fibrillarin, Nor90, Th/To, PM-Scl100, PM-Scl75, Ku, PDGFR, and Ro-52) according to the manufacturer’s recommendations, including the cutoff for seropositivity. Similarly, we used an ANA profile immunoblot by the same manufacturer to detect Scl-70, CENP-B, and PM-Scl. In addition, we determined the erythrocyte sedimentation rate (ESR), C-reactive protein concentration, serum creatinine concentration, the estimated glomerular filtration rate (chronic kidney disease epidemiology collaboration equation; eGFR CKD), as well as the presence or absence of proteinuria.
Statistical analyses were conducted in Statistica 13.3 (TIBCO, Palo Alto, CA, USA) and Microsoft Excel 2016 (Redmond, WA, USA). The associations between autoantibody seropositivity and clinical manifestations or disease subset were assessed using Fisher’s exact test, whereas the comparison of the sensitivity of both immunoblots was made using McNemar’s chi-square test. The significance threshold was set at 0.05. In cases where the contingency table included at least one zero while calculating odds ratios, we used Haldane–Anscombe correction by adding 0.5 to all values in a table so that calculating OR was mathematically possible. Heatmap was created in R 4.1.3 (R Software Foundation, Vienna, Austria) using the pheatmap package 1.0.12, with default hierarchical clustering, after logarithmic transformation (base 10) of odds ratios.
This study obtained the approval of the Bioethics Committee at the Poznan University of Medical Sciences (No. 988/11). All patients participating in the study gave informed consent in writing in line with local law regulations.
Adult
Autoantibodies
Cardiologists
CENPB protein, human
Centromere Protein 17K
Collagen Diseases
Constipation
Contracture
C Reactive Protein
Creatinine
Deglutition Disorders
Diagnosis
Diarrhea
Diastole
Dyspnea
Echocardiography
EGFR protein, human
Europeans
fibrillarin
Finger ulcers
Gender
Glomerular Filtration Rate
Heart
Heart Block
Heartburn
High Blood Pressures
Hypersensitivity
Idiopathic Pulmonary Arterial Hypertension
Immunoblotting
Inpatient
Intestines
Joints
Kidney
Lung
Lung Diseases
Muscle Weakness
Outpatients
Patients
Physical Examination
Physicians
Pulmonologists
Raynaud Phenomenon
Sedimentation Rates, Erythrocyte
Serum
Skin
Synovitis
Systemic Scleroderma
Systole
Tests, Pulmonary Function
Ultrasounds, Doppler
X-Ray Computed Tomography
Protocol full text hidden due to copyright restrictions
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Alkaline Phosphatase
anti-IgG
Antibodies
Antigens
Antigens, Nuclear
Buffers
CENPB protein, human
Diagnosis
DNA, Double-Stranded
Enzymes
Fluorescent Antibody Technique, Indirect
Histones
Homo sapiens
Immunoassay
Immunoblotting
Immunoglobulins
Nucleosomes
Patients
Phosphates
Proliferating Cell Nuclear Antigen
Radionuclide Imaging
Retinitis Pigmentosa 11
Serum
Top products related to «CENPB protein, human»
Sourced in United Kingdom
CENP-B is a protein that localizes to the centromere of chromosomes and plays a role in the organization and function of the centromere. It is involved in the maintenance of centromere structure and the regulation of chromosome segregation during cell division.
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Lipofectamine RNAiMAX is a transfection reagent designed for efficient delivery of small interfering RNA (siRNA) and short hairpin RNA (shRNA) into a wide range of cell types. It is a cationic lipid-based formulation that facilitates the uptake of these nucleic acids into the target cells.
Ab25734 is a laboratory equipment product. It is a device designed for use in scientific research and laboratory settings. The core function of this product is to perform specific tasks or operations required in the laboratory environment. No further details about its intended use or capabilities are provided.
Sourced in United States
Anti-CENP-B is a primary antibody that recognizes the centromere protein B (CENP-B), a protein component of the centromere complex. It can be used in various immunological applications to detect and study the CENP-B protein.
Sourced in United Kingdom
CENP-A is a histone H3 variant that serves as a central component of the centromere-specific nucleosome. It plays a crucial role in the recruitment and assembly of the kinetochore complex, which is essential for proper chromosome segregation during cell division.
Sourced in Germany
The Euroline ANA Profile 3 is a laboratory diagnostic test kit designed to detect and identify autoantibodies associated with various autoimmune disorders. The test utilizes a line assay format to provide qualitative and semi-quantitative results.
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FuGENE HD is a non-liposomal transfection reagent designed for efficient delivery of DNA, RNA, and other macromolecules into a variety of cell types. It offers high transfection efficiency and low cytotoxicity.
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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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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 "CENPB protein, human"
Centromere Protein B (CENPB) is a crucial component of the human centromere, playing a vital role in chromosome segregation and cell division.
This essential protein binds to specific DNA sequences within the centromeric region, contributing to the structural organization and function of this crucial cellular structure.
CENPB has been implicated in various cellular processes, including chromosome stability, kinetochore assembly, and the regulation of gene expression.
Understanding the role of CENPB in human biology can provide important insights into the mechanisms underlying chromosomal abnormalities and related diseases, such as aneuploidy and cancer.
Researchers can leverage PubCompare.ai's AI-powered platform to discover the latest insights on CENP-B, locate optimized research protocols from literature, pre-prints, and patents, and identify the best products for their studies, ultimately boosting reproducibility and streamlining their research efforts.
In addition to CENPB, other key proteins involved in centromere function include CENP-A, which is essential for centromere identity, and CENP-C, which plays a role in kinetochore assembly.
Effective tools for studying CENPB and related proteins include Lipofectamine RNAiMAX for siRNA transfection, Ab25734 for CENP-B detection, and Euroline ANA Profile 3 for autoantibody screening.
Researchers can also utilize FuGENE HD and Lipofectamine 2000 for plasmid transfection, and the RNeasy Mini Kit for RNA extraction and purification.
By leveraging these resources and the insights provided by PubCompare.ai, scientists can advance their understanding of the crucial role of CENPB in human biology and its implications for health and disease.
This essential protein binds to specific DNA sequences within the centromeric region, contributing to the structural organization and function of this crucial cellular structure.
CENPB has been implicated in various cellular processes, including chromosome stability, kinetochore assembly, and the regulation of gene expression.
Understanding the role of CENPB in human biology can provide important insights into the mechanisms underlying chromosomal abnormalities and related diseases, such as aneuploidy and cancer.
Researchers can leverage PubCompare.ai's AI-powered platform to discover the latest insights on CENP-B, locate optimized research protocols from literature, pre-prints, and patents, and identify the best products for their studies, ultimately boosting reproducibility and streamlining their research efforts.
In addition to CENPB, other key proteins involved in centromere function include CENP-A, which is essential for centromere identity, and CENP-C, which plays a role in kinetochore assembly.
Effective tools for studying CENPB and related proteins include Lipofectamine RNAiMAX for siRNA transfection, Ab25734 for CENP-B detection, and Euroline ANA Profile 3 for autoantibody screening.
Researchers can also utilize FuGENE HD and Lipofectamine 2000 for plasmid transfection, and the RNeasy Mini Kit for RNA extraction and purification.
By leveraging these resources and the insights provided by PubCompare.ai, scientists can advance their understanding of the crucial role of CENPB in human biology and its implications for health and disease.