Anti col2a1

Manufactured by Santa Cruz Biotechnology

Sourced in United Kingdom

Anti-COL2A1 is a primary antibody that specifically recognizes the COL2A1 protein. COL2A1 is a gene that encodes the alpha 1 chain of type II collagen, a major structural component of cartilage. This antibody can be used to detect and quantify COL2A1 expression in various biological samples.

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COL2A1 (11 citations) Anti-aggrecan and anti-collagen type II alpha 1 (COL2A1) antibodies (2 citations)

4 protocols using anti col2a1

Immunofluorescent Detection of Chondrogenic Markers

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After washing paraffin sections with PBS, they were incubated in hydrogen peroxidase for 10 min to minimize nonspecific background staining. To detect chondrogenic markers, the sections were incubated with anti-COL2A1, anti-AGGRECAN, and anti-COL10A1 (1:100, Santa Cruz Biotechnology) antibodies at 4 °C for at least 12 h. The attachment of secondary antibodies and fluorescent protein-conjugated secondary antibodies was performed following previously described methods^{59 (link)}. Nuclei were stained with 4ʹ,6-diamidino-2-phenylindole (Sigma). An inverted fluorescence microscope (IX-71; Olympus, Tokyo, Japan) was used to acquire images and expression levels were quantified using ImageJ.

Choi S.M., Lee K.M., Ryu S.B., Park Y.J., Hwang Y.G., Baek D., Choi Y., Park K.H., Park K.D, & Lee J.W. (2018). Enhanced articular cartilage regeneration with SIRT1-activated MSCs using gelatin-based hydrogel. Cell Death & Disease, 9(9), 866.

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Western Blot Analysis of Chondrocyte Markers

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Cell pellets were lysed and quantified as previously reported^{56 (link)}. The samples (10–30 μg of protein) were separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) (Sigma) and transferred onto polyvinylidene difluoride (PVDF) membranes (Amersham Pharmacia, Escondido, CA, USA). Briefly, after blocking with 5% skim milk, membranes were incubated with primary anti-SOX9 (1:3000, Millipore, Billerica, MA, USA), anti-RUNX2, anti-P16 (1:3000, Abcam, Cambridge, UK), anti-COL2A1, anti-AGGRECAN, anti-OCT4, anti-MMP13, anti-COL1A1, anti-P21, anti-P53 (1:500, Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-NANOG (1:1000, BD biosciences, San Jose, CA, USA), anti-SOX2 (1:1000, Cell signaling Technology, Inc., Danver MA, USA), anti-HSP90 or anti-β-ACTIN (1:1,000, Santa Cruz Biotechnology) overnight at 4 °C. Then, membranes were incubated with secondary HRP-conjugated antibodies (1:5000, Santa Cruz Biotechnology) for 1 h at room temperature.

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Evaluating Chondroprotective Effects of High Molecular Weight Hyaluronic Acid

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LPS (Escherichia coli 055: B5) was obtained from Sigma-Aldrich (St. Louis, MO, USA). HMW-HA (0.8–1.5 × 10⁶ Da, 1.8 × 10⁶ Da) was obtain from Meilunbio (Dalian, China). CCK-8 and One Step TUNEL Apoptosis Assay Kit were purchased from Beyotime (Shanghai, China). DAPI solution and FITC conjugated Goat Anti-Rabbit IgG (H + L) was obtained from Servicebio (Wuhan, China). All primary antibodies used in this study were from Proteintech Group (Chicago, IL, USA) except the anti-β-actin (Medical Discovery Leader (MDL) Biotechnology, Beijing, China), anti-IL-1β (Abcam, Cambridge, UK) and anti-COL2A1 (Santa Cruz Biotechnology, Dallas, TX, USA). Bafilomycin A1 (Baf-A1) was purchased from Selleck (Houston, TX, USA). The reagents used for synthesis of o-HA derivatives were of analytical grade.

Huang H., Ding X., Xing D., Lin J., Li Z, & Lin J. (2022). Hyaluronic Acid Oligosaccharide Derivatives Alleviate Lipopolysaccharide-Induced Inflammation in ATDC5 Cells by Multiple Mechanisms. Molecules, 27(17), 5619.

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Histological and Immunohistochemical Assessment of Joint Development

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After sampling for genotyping, the limbs were isolated and immediately fixed in 4% buffered paraformaldehyde for 24 h, decalcified in 14% EDTA (pH 7.4) for 3 days then embedded in paraffin; 6 mm sagittal joint sections were processed for H&E staining.
Immunostaining was performed using a standard protocol as previously described 25 . Primary antibodies were: anti-MINK1 (NO.NBP1-22990, Novus Biologicals), anti-COL2A1 (NO.sc-52658, Santa Cruz Biotechnology Inc.), anti-Aggrecan Neoepitope (NO.NBP100-74350, Novus Biologicals), anti-pSMAD2 (NO.3108, Cell Signaling Technology), anti-osterix (NO.ab22552, Abcam), antiosteocalcin (NO.AB10911, Millipore), anti-VEGF (NO.ab46154, Abcam), anti-MMP13 (NO. ab39012, Abcam), anti-ADAMTS-5 (NO. ab41037, Abcam), anti-COLX (NO.ab58632, Abcam) and anti-nestin (NO.MAB353, Millipore). Stained specimens were photographed digitally under confocal microscopy (BX61W1-FV1000; Olympus) or digital slide scanners (Pannormic MIDI; 3DHISTECH). Mouse IgG or rabbit IgG were used for negative controls when performing immunohistochemistry or immunofluorescence staining (Supplementary Fig. S7).

Yu D., Hu J., Sheng Z., Fu G., Wang Y., Chen Y., Pan Z., Zhang X., Wu Y., Sun H., Dai J., Lu L, & Ouyang H. (2020). Dual roles of misshapen/NIK-related kinase (MINK1) in osteoarthritis subtypes through the activation of TGFβ signaling. Osteoarthritis and cartilage, 28(1).

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