COL2A1 Antibody
Description
Definition and Role of COL2A1 Antibody
The COL2A1 antibody targets the alpha-1 chain of type II collagen, a fibrillar collagen predominantly found in cartilage, the vitreous humor of the eye, and intervertebral discs . Its primary function in research is to study cartilage maintenance, degenerative diseases like osteoarthritis (OA), and genetic disorders such as Stickler syndrome .
Applications in Research
The COL2A1 antibody is validated for use in multiple techniques, including:
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Western blotting (WB): To quantify protein levels in cartilage lysates .
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Immunohistochemistry (IHC): To localize COL2A1 in tissue sections .
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Immunofluorescence (IF/ICC): To visualize collagen synthesis in cultured chondrocytes .
Key Research Findings:
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COL2A1 signaling suppresses chondrocyte hypertrophy, a hallmark of OA progression, by inhibiting BMP-SMAD1 pathways .
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Antibody-mediated detection of COL2A1 degradation correlates with cartilage erosion in OA models .
Therapeutic and Diagnostic Implications
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Osteoarthritis: COL2A1 antibodies are used to monitor collagen degradation in joint cartilage, aiding in early OA diagnosis .
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Regenerative Medicine: Studies with COL2A1 antibodies have identified pathways promoting cartilage repair in mesenchymal stem cells .
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Genetic Disorders: Mutations in COL2A1 linked to Stickler syndrome and achondrogenesis are detectable via antibody-based assays .
Product Specs
Target Background
Type II collagen is a cartilage-specific protein crucial for normal skeletal embryonic development, linear growth, and cartilage's resistance to compressive forces.
COL2A1 Gene References and Associated Functions
- Differentiating Stickler Syndrome: Routine ocular exams may not easily distinguish Stickler syndrome (STL) from early-onset high myopia. Recognizing atypical phenotypes and newly identified signs aids in atypical STL identification, particularly in pediatric ophthalmology. PMID: 30181686
- Expanding the COL2A1 Mutation Spectrum: This study expands the known COL2A1 mutation spectrum, potentially improving genetic counseling and therapeutic strategy development for Stickler syndrome patients. PMID: 30015854
- COL2A1 and Kashin-Beck Disease: Findings suggest COL2A1's potential role as a susceptibility gene for Kashin-Beck disease (KBD), possibly influencing KBD-related hand growth and development impairments. PMID: 28059113
- COL2A1 Mutation and Avascular Necrosis: The COL2A1 mutation (c.3508G>A) is linked to avascular necrosis of the femoral head in a Chinese family. PMID: 29750297
- Misfolded COL2A1 Proteins and ER Stress: Retention of misfolded R740C and R789C proteins triggers endoplasmic reticulum (ER) stress. These proteins exhibit significantly reduced melting temperatures. PMID: 29439465
- COMP Mutation and Intracellular Protein Accumulation: A novel COMP mutation may result in mutant protein intracellular accumulation. Decreased plasma COMP and increased plasma CTX-II might serve as diagnostic markers for pseudoachondroplasia (PSACH), though their presymptomatic applicability remains unclear. PMID: 29104872
- Type II Collagen Degradation and Vitamin D Receptor Polymorphisms: Higher circulating Type II Collagen Degradation Marker (CTx-II) levels were observed in patients with specific VDR polymorphisms (bb, Aa, TT genotypes; F and T alleles) compared to healthy controls in the context of osteochondrosis. PMID: 28961166
- COL2A1 Mutation and Metaphyseal Abnormalities: A c.G1636A (p.G546S) COL2A1 mutation in a family was associated with varying metaphyseal changes. This glycine-to-serine substitution is linked to "dappling" and "corner fracture" metaphyseal abnormalities, offering insights into the phenotypic spectrum of type II collagenopathies. PMID: 28738883
- Novel COL2A1 Mutations in Chinese Stickler Syndrome Patients: Three novel and two known COL2A1 gene mutations were identified in Chinese Stickler syndrome patients, expanding the known mutation spectrum. PMID: 27390512
- Novel COL2A1 Mutations in Spondyloepiphyseal Dysplasia Congenita: Three novel heterozygous COL2A1 mutations (Gly537Asp, Gly909Ser, and Gly1149Val) were identified in unrelated Chinese families with Spondyloepiphyseal dysplasia congenita. PMID: 27059630
- Integrin alpha10beta1 and Collagen Recognition: Unique charge-dependent constraints on collagen recognition by integrin alpha10beta1 have been described. PMID: 27569273
- Novel Nonsense Mutation in COL2A1: A novel nonsense mutation in COL2A1 exon 2 shows incomplete penetrance and/or variable age of onset with extraocular manifestations. PMID: 28095098
- COL2A1 Intron 2 Variant and Splicing: This report examines how a COL2A1 intron 2 de novo variant and polymorphism affect exon 2 inclusion in the COL2A1 transcript and identifies potential trans-acting splicing factors. A significant difference in the frequency of the COL2A1 variant rs1635532 was found between patients with rhegmatogenous retinal detachment and controls. PMID: 27406592
- COL2A1 Testing in Spondylometaphyseal Dysplasia: Given the overlap between spondylometaphyseal dysplasia (SMD) corner fracture type and type II collagenopathies, COL2A1 molecular testing is warranted in patients with corner fracture-like lesions in the context of SMD. PMID: 27888646
- Type II Collagen Expression and Intervertebral Disc Degeneration: High type II collagen expression is associated with intervertebral disc degeneration. PMID: 28559201
- Endoplasmic Reticulum Stress and Osteoarthritic Chondrocytes: ER stress contributes to senescence and apoptosis of osteoarthritic chondrocytes, characterized by increased ADAMTS5 and MMP13 expression and decreased COL2A1 expression. PMID: 28728848
- ELF3 and COL2A1 Transcriptional Regulation: This study highlights ELF3's contribution to COL2A1 transcriptional regulation. PMID: 27310669
- COL2A1 Variants in Stickler Syndrome: Approximately 46% of Stickler syndrome patients carried a COL2A1 variant, with varying molecular spectra across different phenotypes. PMID: 26626311
- SOX9/5/6 and Col2a1 Transcription: The SOX9/5/6 combination enhances Col2a1 transcription through novel enhancers in introns 1 and 6. PMID: 27881681
- COL2A1 Mutation Analysis in Idiopathic Osteonecrosis: Analysis of COL2A1 mutations in idiopathic osteonecrosis of the femoral head was conducted among patients from 22 Japanese hospitals. PMID: 27183340
- COL2A1 and Aggrecan Polymorphisms in Intervertebral Disc Degeneration: Genotype and allele frequencies of COL2A1 polymorphisms (rs1793953 and rs2276454) and Aggrecan VNTR polymorphisms differed significantly between patients with intervertebral disc degeneration and controls, particularly at higher Pfirrmann grades (III, IV, V). PMID: 27991836
- miR-133a and Type II Collagen Loss in Intervertebral Disc Degeneration: Type II collagen loss is induced by miR-133a downregulation in intervertebral disc degeneration. PMID: 26656045
- COL2A1 mRNA Expression and Osteoarthritis: Decreased COL2A1 mRNA expression is associated with osteoarthritis. PMID: 27428952
- COL2A1 Defects in Osteoarthritis: COL2A1 defects in osteoarthritis are not limited to exon 2 mutations; c.2678dupC (p.Ala895Serfs*49) and c.3327+1G>C are examples. PMID: 26709265
- COL2A1 Mutations and Bone Dysplasia: Mutations in COL2A1 cause a wide clinical spectrum of bone dysplasias, ranging from lethal skeletal abnormalities to isolated arthritis. PMID: 26443184
- miR-27b and Type II Collagen Expression: miR-27b overexpression promotes type II collagen expression in nucleus pulposus cells. PMID: 26583473
- Novel COL2A1 Mutation and Mild Spondyloepiphyseal Dysplasia: Whole exome sequencing identified a novel COL2A1 mutation causing mild spondyloepiphyseal dysplasia, mimicking autosomal dominant brachyolmia. PMID: 26586363
- Dysspondyloenchondromatosis and COL2A1 Mutation: Dysspondyloenchondromatosis is associated with COL2A1 mutation. PMID: 26250472
- COL2A1 Mutation and Kniest Dysplasia/Chondrosarcoma: Constitutive COL2A1 gene mutations are associated with Kniest dysplasia and chondrosarcoma. PMID: 26345137
- Serum CTX-II Levels in Human Brucellosis: Serum CTX-II levels were higher in human brucellosis patients than in healthy controls, with significantly higher levels in males, suggesting cartilage and bone changes. PMID: 27109135
- Novel COL2A1 Variant and Type II Collagenopathy: A novel COL2A1 variant (c.619G>A, p.Gly207Arg) causes a distinct type II collagenopathy with features of progressive pseudorheumatoid dysplasia and spondyloepiphyseal dysplasia, Stanescu type. PMID: 26183434
- COL2A1 and SLC6A10P Expression and Disease Progression: Subjects expressing certain levels of COL2A1 and SLC6A10P showed a median disease progression of 95 months, compared to 16 months for other expression levels. PMID: 26311224
- Novel COL2A1 Mutation and Spondyloepiphyseal Dysplasia Congenita: A novel mutation, c.620G>A (p.Gly207Glu), in the collagen type II alpha-1 gene, is associated with spondyloepiphyseal dysplasia congenita; genotype-phenotype relationships are discussed. PMID: 26030151
- Col2a1 Mutant Mouse Model and Platyspondylic Lethal Skeletal Dysplasia: A novel Col2a1 mutant mouse (p.Tyr1391Ser) displays skeletal dysplasia resembling platyspondylic lethal skeletal dysplasia, Torrance type, due to ER stress-mediated apoptosis. PMID: 26545783
- Double De Novo COL2A1 Mutations and Spondyloepiphyseal Dysplasia Congenita: A unique case of spondyloepiphyseal dysplasia congenita with mild coxa vara is caused by double de novo COL2A1 mutations (p.G504S, p.G612A) on the same allele. PMID: 25900302
- Novel COL2A1 Mutation and Spondyloepiphyseal Dysplasia Congenita: c.2224G>A (p.Gly687Ser) is a novel COL2A1 mutation associated with spondyloepiphyseal dysplasia congenita. PMID: 25967556
- R989C COL2A1 Mutation and Severe Spondyloepiphyseal Dysplasia Congenita: Six unrelated patients with the R989C COL2A1 mutation exhibited a severe spondyloepiphyseal dysplasia congenita phenotype. PMID: 25735649
- Urine C2C and Trace Element Levels in Knee Osteoarthritis: Analysis of urine C2C and trace element levels in patients with knee osteoarthritis. PMID: 24728947
- Novel COL2A1 Mutation and Kniest Dysplasia: A novel missense mutation (c.905C>T, p.Ala302Val) in COL2A1 is associated with Kniest dysplasia. PMID: 26037341
- miR-93, MMP3, and Type II Collagen Expression: miR-93 affects nucleus pulposus cell type II collagen expression by targeting MMP3, implicated in intervertebral disc degeneration. PMID: 25818544
- Novel COL2A1 Mutation and Spondyloepiphyseal Dysplasia Congenita in a Chinese Family: A novel missense mutation, c.2224G>A (p.Gly687Ser), in COL2A1 is associated with spondyloepiphyseal dysplasia congenita in a Chinese family. PMID: 25863096
- COL2A1 Somatic Alterations in Chondrosarcoma and Enchondroma: Somatic COL2A1 alterations were found in 19.3% of chondrosarcoma and 31.7% of enchondroma cases. PMID: 25024164
- COL2A1 and Degenerative Lumbar Scoliosis: COL2A1 is associated with the risk of degenerative lumbar scoliosis in the Korean population. PMID: 25436060
- COL2A1 Mutation and Osteonecrosis of the Femoral Head: A COL2A1 mutation causes osteonecrosis of the femoral head in a family. PMID: 25050885
- Genetic and Environmental Influences on Collagen IIA Synthesis: Approximately 45% of collagen IIA synthesis (serum collagen IIA N-terminal propeptide) is attributable to genetics, while individual and shared environments contribute 24% and 31%, respectively. PMID: 25008205
- T Cell Specificity Antibody to Type II Collagen: Data indicate T cell specificity antibody to the CII259-273 T cell epitope in B10.DR4.Ncf1*/* mice following immunization with human type II collagen (CII). PMID: 23116329
- COL2A1 Mutations and Type II Collagenopathies: Mutations in COL2A1 cause a range of type II collagenopathies, manifesting as inheritable skeletal disorders. PMID: 25124518
- COL2A1 Polymorphism and Multicystic Dysplastic Kidney: An association was observed between COL2A1 gene polymorphisms and multicystic dysplastic kidney (MP), suggesting COL2A1 as a potential susceptibility gene. PMID: 24386886
- uCTX-II Levels and Knee Structural Changes: Higher urinary C-terminal telopeptide of type II collagen (uCTX-II) levels in middle-aged women without clinical knee disease were associated with early knee structural changes (cartilage defects, tibial bone expansion, bone marrow lesions) at baseline. PMID: 24971869
- Novel COL2A1 Variants and Collagenopathies: New variants in COL2A1 resulting in related collagenopathies are described. PMID: 24949742
Customer Reviews
Applications : Immunoblotting
Sample type: cells
Review: IL-1β significantly decreased aggrecan,collagen II, SOX9, and FN1 mRNA and protein levels.
Q&A
What is COL2A1 and where is it primarily expressed?
COL2A1 encodes the alpha-1 chain of type II collagen, a fibrillar collagen that forms the structural framework of cartilaginous tissues. It is predominantly expressed in chondrocytes and is essential for normal skeletal development and cartilage function . COL2A1 is also found in the vitreous humor of the eye . Recent research has identified expression in additional tissues including embryonic stem cell muscle, fetal sternum, and blood according to multiple studies . The protein is primarily localized to the extracellular matrix and extracellular space .
Which COL2A1 antibody type should I select for my experiment?
The choice between monoclonal and polyclonal antibodies depends on your specific research goals:
Polyclonal antibodies (such as A00517-1, bs-10589R):
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Recognize multiple epitopes, providing stronger signal amplification
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Ideal for detecting proteins in native conformation
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Better for immunoprecipitation and some ELISA formats
Monoclonal antibodies (such as sc-518017):
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Recognize a single epitope, offering higher specificity
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More consistent lot-to-lot reproducibility
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Better for distinguishing closely related proteins
For detecting specific COL2A1 isoforms (IIA vs. IIB), specialized antibodies targeting the exon junction regions are recommended .
What is the observed molecular weight of COL2A1 and why might it vary?
While the calculated molecular weight of COL2A1 is approximately 142 kDa , Western blot often reveals bands at 180-190 kDa . This discrepancy is due to:
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Post-translational modifications (particularly glycosylation)
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The presence of pro-collagen domains in unprocessed forms
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Different isoforms due to alternative splicing (IIA vs. IIB)
When running Western blots, it's advisable to use 5-20% gradient gels to properly resolve this high molecular weight protein .
What are the recommended dilutions for different applications?
Based on multiple manufacturers' guidelines:
| Application | Recommended Dilution | Reference |
|---|---|---|
| Western blot | 1:300-5000 or 0.1-0.5 μg/ml | |
| IHC-Paraffin | 1:200-5000 | |
| IHC-Frozen | 0.5-1 μg/ml | |
| Immunofluorescence | 1:50-200 | |
| ELISA | 1:500-50,000 | |
| Immunoprecipitation | 1:100 |
Always perform titration experiments to determine optimal concentration for your specific sample and conditions .
Which positive controls are recommended for validating COL2A1 antibody specificity?
For reliable validation, the following positive controls have been verified across multiple studies:
Tissue controls:
Negative controls:
Including both positive and negative controls is essential for confirming antibody specificity and optimizing experimental conditions .
What is the optimal sample preparation protocol for COL2A1 Western blotting?
For successful detection of COL2A1 by Western blot:
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Sample preparation:
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Electrophoresis conditions:
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Transfer conditions:
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Antibody incubation:
This protocol has been validated with COL2A1 antibodies showing specific bands at approximately 180-190 kDa .
How can COL2A1 antibodies be used to study cartilage development and disease models?
COL2A1 antibodies are powerful tools for investigating cartilage development and pathology:
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Developmental studies:
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Disease models:
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Regenerative medicine applications:
How are COL2A1 antibodies utilized in tumor biology research?
Recent studies have revealed unexpected roles for COL2A1 in cancer:
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Melanoma tumor repopulating cells (TRCs):
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COL2A1 expression is significantly upregulated (17-20 fold) in melanoma TRCs compared to parental B16F1 control cells
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Immunofluorescence microscopy with anti-COL2A1 antibodies confirms approximately 14-fold higher signal in TRCs
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The expression pattern correlates with metastatic potential, with higher expression in more metastatic B16F10 cells compared to B16F1 cells
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Potential biomarker applications:
This represents a novel application area where COL2A1 antibodies can contribute to understanding cancer stem cell biology and potential therapeutic targets .
What techniques can be used to distinguish between type IIA and IIB procollagen isoforms?
Distinguishing between COL2A1 isoforms requires specialized approaches:
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Isoform-specific antibodies:
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Experimental models:
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Combined approaches:
These techniques allow researchers to track chondrocyte differentiation and study developmental processes in cartilage formation.
Why might I observe unexpected tissue staining with COL2A1 antibodies?
Unexpected COL2A1 expression in non-cartilaginous tissues may occur due to:
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Genuine expression in non-canonical tissues:
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Cross-reactivity issues:
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Technical considerations:
When observing unexpected staining, it's advisable to confirm findings using alternative detection methods such as RT-PCR or in situ hybridization .
What causes multiple bands in Western blot analysis of COL2A1?
Multiple bands in COL2A1 Western blots can result from:
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Protein processing stages:
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Alternative splicing:
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Degradation products:
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Proteolytic cleavage during sample preparation
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Disease-associated degradation (e.g., in osteoarthritis samples)
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Technical issues:
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Sample overheating during preparation
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Insufficient reducing conditions
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Incomplete denaturation of the triple-helical structure
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To minimize these issues, use freshly prepared samples with protease inhibitors, proper reducing agents, and optimize gel concentration based on the expected molecular weight range .
How can I validate antibody specificity and address cross-reactivity concerns?
To ensure antibody specificity and minimize cross-reactivity:
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Comprehensive validation approach:
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Cross-adsorption techniques:
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Specificity testing:
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ELISA testing against purified standards of different collagen types
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Western blot analysis of tissues with known expression patterns
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Peptide competition assays to confirm epitope specificity
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High-quality COL2A1 antibodies should show negligible cross-reactivity (<1%) with collagen types I, III, IV, V, or VI .
These validation steps are essential for generating reliable data and avoiding misinterpretation of results, particularly in tissues with complex extracellular matrix composition.
