Product: Collagen II Antibody
Catalog: AF0135
Description: Rabbit polyclonal antibody to Collagen II
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Bovine, Horse, Sheep, Rabbit, Chicken, Xenopus
Mol.Wt.: 140kDa, 30~50kd(possible isoform); 142kD(Calculated).
Uniprot: P02458
RRID: AB_2833318

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:3000, IHC 1:50-1:200, IF/ICC: 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Chicken(86%), Xenopus(86%)
Clonality:
Polyclonal
Specificity:
Collagen II Antibody detects endogenous levels of total Collagen II.
RRID:
AB_2833318
Cite Format: Affinity Biosciences Cat# AF0135, RRID:AB_2833318.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

Alpha 1 type II collagen;Alpha-1 type II collagen;AOM;Cartilage collagen;Chondrocalcin;CO2A1_HUMAN;COL11A3;Col2a1;Collagen II alpha 1 polypeptide;SEDC;col 2;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
P02458 CO2A1_HUMAN:

Isoform 2 is highly expressed in juvenile chondrocyte and low in fetal chondrocyte.

Description:
COL2A1 Type II collagen is specific for cartilaginous tissues. It is essential for the normal embryonic development of the skeleton, for linear growth and for the ability of cartilage to resist compressive forces. Belongs to the fibrillar collagen family. Homotrimers of alpha 1(II) chains. 3 isoforms of the human protein are produced by alternative splicing
Sequence:
MIRLGAPQTLVLLTLLVAAVLRCQGQDVQEAGSCVQDGQRYNDKDVWKPEPCRICVCDTGTVLCDDIICEDVKDCLSPEIPFGECCPICPTDLATASGQPGPKGQKGEPGDIKDIVGPKGPPGPQGPAGEQGPRGDRGDKGEKGAPGPRGRDGEPGTPGNPGPPGPPGPPGPPGLGGNFAAQMAGGFDEKAGGAQLGVMQGPMGPMGPRGPPGPAGAPGPQGFQGNPGEPGEPGVSGPMGPRGPPGPPGKPGDDGEAGKPGKAGERGPPGPQGARGFPGTPGLPGVKGHRGYPGLDGAKGEAGAPGVKGESGSPGENGSPGPMGPRGLPGERGRTGPAGAAGARGNDGQPGPAGPPGPVGPAGGPGFPGAPGAKGEAGPTGARGPEGAQGPRGEPGTPGSPGPAGASGNPGTDGIPGAKGSAGAPGIAGAPGFPGPRGPPGPQGATGPLGPKGQTGEPGIAGFKGEQGPKGEPGPAGPQGAPGPAGEEGKRGARGEPGGVGPIGPPGERGAPGNRGFPGQDGLAGPKGAPGERGPSGLAGPKGANGDPGRPGEPGLPGARGLTGRPGDAGPQGKVGPSGAPGEDGRPGPPGPQGARGQPGVMGFPGPKGANGEPGKAGEKGLPGAPGLRGLPGKDGETGAAGPPGPAGPAGERGEQGAPGPSGFQGLPGPPGPPGEGGKPGDQGVPGEAGAPGLVGPRGERGFPGERGSPGAQGLQGPRGLPGTPGTDGPKGASGPAGPPGAQGPPGLQGMPGERGAAGIAGPKGDRGDVGEKGPEGAPGKDGGRGLTGPIGPPGPAGANGEKGEVGPPGPAGSAGARGAPGERGETGPPGPAGFAGPPGADGQPGAKGEQGEAGQKGDAGAPGPQGPSGAPGPQGPTGVTGPKGARGAQGPPGATGFPGAAGRVGPPGSNGNPGPPGPPGPSGKDGPKGARGDSGPPGRAGEPGLQGPAGPPGEKGEPGDDGPSGAEGPPGPQGLAGQRGIVGLPGQRGERGFPGLPGPSGEPGKQGAPGASGDRGPPGPVGPPGLTGPAGEPGREGSPGADGPPGRDGAAGVKGDRGETGAVGAPGAPGPPGSPGPAGPTGKQGDRGEAGAQGPMGPSGPAGARGIQGPQGPRGDKGEAGEPGERGLKGHRGFTGLQGLPGPPGPSGDQGASGPAGPSGPRGPPGPVGPSGKDGANGIPGPIGPPGPRGRSGETGPAGPPGNPGPPGPPGPPGPGIDMSAFAGLGPREKGPDPLQYMRADQAAGGLRQHDAEVDATLKSLNNQIESIRSPEGSRKNPARTCRDLKLCHPEWKSGDYWIDPNQGCTLDAMKVFCNMETGETCVYPNPANVPKKNWWSSKSKEKKHIWFGETINGGFHFSYGDDNLAPNTANVQMTFLRLLSTEGSQNITYHCKNSIAYLDEAAGNLKKALLIQGSNDVEIRAEGNSRFTYTALKDGCTKHTGKWGKTVIEYRSQKTSRLPIIDIAPMDIGGPEQEFGVDIGPVCFL

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Horse
100
Bovine
100
Sheep
100
Rabbit
100
Xenopus
86
Chicken
86
Dog
67
Pig
0
Zebrafish
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P02458 As Substrate

Site PTM Type Enzyme
Y41 Phosphorylation
K490 Ubiquitination
S536 Phosphorylation
K542 Acetylation
K764 Acetylation
K773 Acetylation
T1307 Phosphorylation
S1416 Phosphorylation

Research Backgrounds

Function:

Type II collagen is specific for cartilaginous tissues. It is essential for the normal embryonic development of the skeleton, for linear growth and for the ability of cartilage to resist compressive forces.

PTMs:

The N-telopeptide is covalently linked to the helical COL2 region of alpha 1(IX), alpha 2(IX) and alpha 3(IX) chain. The C-telopeptide is covalently linked to an another site in the helical region of alpha 3(IX) COL2.

Contains mostly 4-hydroxyproline. Prolines at the third position of the tripeptide repeating unit (G-X-P) are 4-hydroxylated in some or all of the chains.

Contains 3-hydroxyproline at a few sites. This modification occurs on the first proline residue in the sequence motif Gly-Pro-Hyp, where Hyp is 4-hydroxyproline.

Lysine residues at the third position of the tripeptide repeating unit (G-X-Y) are 5-hydroxylated in some or all of the chains.

O-glycosylated on hydroxylated lysine residues. The O-linked glycan consists of a Glc-Gal disaccharide.

Subcellular Location:

Secreted>Extracellular space>Extracellular matrix.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Isoform 2 is highly expressed in juvenile chondrocyte and low in fetal chondrocyte.

Subunit Structure:

Homotrimers of alpha 1(II) chains.

Family&Domains:

The C-terminal propeptide, also known as COLFI domain, have crucial roles in tissue growth and repair by controlling both the intracellular assembly of procollagen molecules and the extracellular assembly of collagen fibrils. It binds a calcium ion which is essential for its function (By similarity).

Belongs to the fibrillar collagen family.

Research Fields

· Cellular Processes > Cellular community - eukaryotes > Focal adhesion.   (View pathway)

· Environmental Information Processing > Signal transduction > PI3K-Akt signaling pathway.   (View pathway)

· Environmental Information Processing > Signaling molecules and interaction > ECM-receptor interaction.   (View pathway)

· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.

· Organismal Systems > Digestive system > Protein digestion and absorption.

References

1). Hu B et al. HSP70 attenuates compression-induced apoptosis of nucleus pulposus cells by suppressing mitochondrial fission via upregulating the expression of SIRT3. EXPERIMENTAL AND MOLECULAR MEDICINE 2022 Mar;54(3):309-323. (PubMed: 35338257) [IF=12.8]

2). Kou L et al. Opsonized nanoparticles target and regulate macrophage polarization for osteoarthritis therapy: A trapping strategy. Journal of Controlled Release 2022 Jul;347:237-255. (PubMed: 35489544) [IF=10.8]

3). Zheng K et al. Co-culture pellet of human Wharton’s jelly mesenchymal stem cells and rat costal chondrocytes as a candidate for articular cartilage regeneration: in vitro and in vivo study. Stem Cell Research & Therapy 2022 Jul 30;13(1):386. (PubMed: 35907866) [IF=7.5]

Application: IHC    Species: Rat    Sample:

Fig. 3Histological and immunochemistry staining of pellets with semiquantitative analysis. A H–E and Safranin-O staining of pellets and partial enlargement in five groups. B Immunochemistry staining of COLII and COLX of pellets and partial enlargement in five groups. C–E Semiquantitative analysis of AOI of each staining (n = 4). Significant difference symbols: *p < 0.05, **p < 0.01 compared to CCs group, #p < 0.05, ##p < 0.01 compared to hWJMSCs group

4). Zhao R et al. Lysyl oxidase inhibits TNF-α induced rat nucleus pulposus cell apoptosis via regulating Fas/FasL pathway and the p53 pathways. LIFE SCIENCES 2020 Sep 23;118483. (PubMed: 32979358) [IF=6.1]

Application: WB    Species: rat    Sample: NP cells

Fig. 3. |LOX improved the ECM expression pattern in TNF-α-treated rat NP cells. A. and B. Relative mRNA expressions of type II collagen and aggrecan in rat NP cells with different treatments were analyzed by RT-qPCR. C. Protein expressions of type II collagen and aggrecan in rat NP cells with different treatments were analyzed by western blot.

5). Zhou J et al. MicroRNA-145 overexpression attenuates apoptosis and increases matrix synthesis in nucleus pulposus cells. LIFE SCIENCES 2019 Feb 20 (PubMed: 30797016) [IF=6.1]

Application: WB    Species: rat    Sample: NP cells

Fig. 3. |miR-145 increased matrix metabolism in rat NP cells.(A, B) Real-time PCR analysis was performed to evaluate the mRNA expression of aggrecan (A) and collagen IIa (B) after a single transfection with agomir-145 or miR-155 inhibitors. (C, D, E) Western blotting (C) and subsequent densitometric analysis (D) demonstrated that aggrecan (C, E) and collagen IIa (D, E) protein were significantly upregulated in rat NP cells after transfection with agomir-145.

6). Ma X et al. L-Glutamine alleviates osteoarthritis by regulating lncRNA-NKILA expression through the TGF-β1/SMAD2/3 signalling pathway. Clinical Science 2022 Jun 22; (PubMed: 35730575) [IF=6.0]

7). Zheng X et al. PD184352 exerts anti-inflammatory and antioxidant effects by promoting activation of the Nrf2/HO-1 axis. Biochemical Pharmacology 2023 Apr 05; (PubMed: 37028460) [IF=5.8]

8). Wei X et al. Nerve growth factor promotes ASIC1a expression via the NF-κB pathway and enhances acid-induced chondrocyte apoptosis. International Immunopharmacology 2020 Mar 5;82:106340 (PubMed: 32146316) [IF=5.6]

Application: IHC    Species: rat    Sample: articular chondrocytes

Fig. 1. |NGF up-regulated ASIC1a expression in rat articular chondrocytes. (A) Extracorporeal culture and identification of primary articular chondrocytes in rats. (i) The biosynthesis of intracellular glycosaminoglycan stained positive with toluidine blue. (ii) Immunohistochemical staining of collagen.

9). Guo Q et al. PDLIM2 protects articular chondrocytes from lipopolysaccharide-induced apoptosis, degeneration and inflammatory injury through down-regulation of nuclear factor (NF)-κB signaling. International Immunopharmacology 2020 Aug 14;88:106883. (PubMed: 32805696) [IF=5.6]

Application: WB    Species: Mouse    Sample: chondrocytes

Fig. 2. Up-regulation of PDLIM2 attenuates LPS-induced injury in chondrocytes. Chondrocytes were transfected with empty vector (EV) or PDLIM2 expression vector for 48 h and then stimulated with 10 μg/ml of LPS for 6 h. (A) The relative mRNA expression of PDLIM2 was determined by RT-qPCR. (B, C) Protein expression of PDLIM2 was examined by Western blotting. (D) The effect of PDLIM2 overexpression on cell viability was detected by MTT assay. The effect of PDLIM2 overexpression on cell apoptosis was assessed by (E, F) TUNENL and (G, H) Annexin V-FITC/PI apoptosis assays. (I) The effect of PDLIM2 overexpression on MMP-3, MMP-13, COL2A1 and ACAN was evaluated by Western blotting. Quantitative analysis of protein expression of (J) MMP-3, (K) MMP-13, (L) COL2A1 and (M) ACAN. N = 3, **p < 0.01 and ***p < 0.001.

10). Min L et al. A network pharmacology strategy to investigate the anti-osteoarthritis mechanism of main lignans components of Schisandrae Fructus. International Immunopharmacology 2021 Jun 25;98:107873. (PubMed: 34182246) [IF=5.6]

Application: WB    Species: Human    Sample: SW1353 cells

Fig. 6. (A) Analysis of the cytotoxicity induced by the 6 lignans in SW1353 cells. (B) Effects of the 6 lignans on collagen II gene expression in IL-1βtreated SW1353 cells. (C) Effects of the 6 lignans on MMP13 gene expression in IL-1β-treated SW1353 cells. (D) Effects of schisanhenol and gammaschisandrin on collagen II gene expression in IL-1βtreated SW1353 cells. (E) Effect of schisanhenol on MMP13 and collagen II protein expression in IL-1βtreated SW1353 cells. All the data are represented as the mean ± SD. Ns represents not significant, *p < 0.05, **p < 0.01 and ***p < 0.001, versus the control group; #p < 0.05, ##p < 0.01 and ###p < 0.001, versus the IL-1β-treated group.

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