Collagen 2

Manufactured by Affinity Biosciences

Sourced in United States, China

Collagen II is a type of collagen found in the cartilage and other connective tissues. It is a structural protein that provides strength and flexibility to these tissues. The product from Affinity Biosciences is a purified form of Collagen II, suitable for a variety of research and laboratory applications.

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Lab products found in correlation

Mmp13, by Proteintech (2 mentions) Bcl2 associated x (bax), by Proteintech (2 mentions) 4 6 diamidino 2 phenylindole dapi, by Abcam (1 mentions) Aggrecan, by Abcam (1 mentions) Tom20, by Cell Signaling Technology (1 mentions) Collagen 2, by Cell Signaling Technology (1 mentions) Erbb2, by Affinity Biosciences (1 mentions) Ptgs2, by Affinity Biosciences (1 mentions) Triton x 100, by Beyotime (1 mentions) Fluorescence microscope, by Olympus (1 mentions) Bovine serum albumin (bsa), by Sangon (1 mentions) Paraformaldehyde (pfa), by Sinopharm (1 mentions) Cy3 conjugated goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions) Ripa lysis buffer, by Beyotime (1 mentions) Pvdf membrane, by Merck Group (1 mentions) Bcl 2, by Abcam (1 mentions) β actin, by Cell Signaling Technology (1 mentions) Aggrecan, by Novus Biologicals (1 mentions) Caspase 9, by Proteintech (1 mentions) Ecl kit, by Affinity Biosciences (1 mentions)

Close spelling variants of this product

Rabbit antibodies against collagen type II (2 citations)

5 protocols using collagen 2

Immunostaining of Bovine Nucleus Pulposus Cells

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NPCs were fixed on coverslips with 4% paraformaldehyde for 20 min, then permeabilized and blocked with PBS containing 0.3% Triton X-100 and 5% BSA for 1 h at room temperature. The slides were then incubated with primary antibodies collagen II (1:200, Cell Signaling Technology), Aggrecan (1:200, Abcam), Tom20 (1:200, Cell Signaling Technology), LC3 (1:200, Cell Signaling Technology) at 4°C overnight. After washing with TBST, slides were incubated with Alexa Fluor-488 or Fluor-594 conjugated secondary antibody (1:300, Jackson ImmunoResearch Inc., United States) for 1 h at room temperature and labeled with 4′,6-Diamidino-2-Phenylindole (DAPI, Abcam) for 5 min. Finally, the staining was visualized in a confocal fluorescence microscope (Zeiss Confocal LSM 780, Germany) and fluorescence intensity was measured using ImageJ software.
Bovine caudal NP tissues were fixed in 4% paraformaldehyde for 24 h and dehydrated in graded sucrose solutions. Then, the tissues were embedded in OCT compound (Sakura, United States), and cut into 10 µm-thick cryosections. The following permeabilization, block and immunostaining protocols were identical to coverslips as described above. These following primary antibodies were used: Bax (1:200, Proteintech), collagen II (1:200, Affinity, United States), Mmp13 (1:200, Proteintech).

Zhang F., Wang S., Gao M., Li B., He Z., Tang T., Zhu Z., Liu S, & Zhou Z. (2022). Hyaluronic acid ameliorates intervertebral disc degeneration via promoting mitophagy activation. Frontiers in Bioengineering and Biotechnology, 10, 1057429.

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Histological and Immunohistochemical Evaluation of Cartilage

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The paraffin-embedded discs were cut into 4 μm-thick sections. For histology, the sections were deparaffinized, rehydrated, and then stained with hematoxylin and eosin (HE) and Safranin O-Fast Green (SO). In addition, the histopathological grading scores were assessed according to the grading scale [39 (link)]. For IHC staining, the sections were deparaffinized, rehydrated, and incubated with 3% H₂O₂ for 20 minutes at room temperature to eliminate endogenous peroxidase activity. Then, the antigen was retrieved by pressure cooking in 10 mmol/L citrate buffer, pH = 6. Subsequently, the sections were blocked with 10% goat serum albumin for 30 min at room temperature. The sections were then incubated with a primary antibody against Nrf2 (1 : 400, ProteinTech), collagen II (1 : 50, Affinity Biosciences), and MMP13 (1 : 300, ProteinTech) at 4°C overnight. Sections incubated with nonspecific IgG were used as negative controls. Next, the sections were incubated with horseradish peroxidase-conjugated goat anti-rabbit secondary antibody for 1 h at 37°C and counterstained with hematoxylin. For the gray-scale statistics of IHC staining, we used ImageJ to calculate the integrated optical density (IOD) of IHC images.

Wang P., Zhang S., Liu W., Chen S., Lv X., Hu B, & Shao Z. (2022). Selenium Attenuates TBHP-Induced Apoptosis of Nucleus Pulposus Cells by Suppressing Mitochondrial Fission through Activating Nuclear Factor Erythroid 2-Related Factor 2. Oxidative Medicine and Cellular Longevity, 2022, 7531788.

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Immunofluorescence Assay for Chondrocyte Markers

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The third-generation human NP cells was tested, remove the culture medium, wash twice with PBS buffer, incubate with 4% neutral paraformaldehyde fixative solution for 5 min at room temperature; wash twice with PBS buffer, 0.1% TritonX-100 cells were incubated with permeabilization solution at room temperature for 20 min; goat serum was blocked for 1 h; blocking solution was removed, Collagen II(1:200), Agreecan (1:200), ERBB2(1:150), PTGS2(1:200),and P16(1:200) (Affinity Biosciences, China) were added, and incubated overnight at 4 °C; primary antibody was removed and washed with PBS for 5 min×3 times, add fluorescent secondary antibody working solution and incubate for 1 h; discard the secondary antibody, wash with PBS for 5 min×3 times, add DAPI staining working solution and incubate for 20 min; observe under an inverted fluorescence microscope and take pictures for recording.

Li Z.C, & An F. (2023). ERBB2-PTGS2 axis promotes intervertebral disc degeneration by regulating senescence of nucleus pulposus cells. BMC Musculoskeletal Disorders, 24, 504.

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Chondrocyte Immunofluorescence Staining

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The chondrocytes were fixed with 4% paraformaldehyde (Sinopharm, Shanghai, China) for 15 min, permeabilized with 0.1% Triton X-100 (Beyotime, Shanghai, China) for 30 min, and blocked by 1% bovine serum albumin (Sangon, Shanghai, China) for 15 min. The samples were incubated with collagen II (1:100; Affinity, Changzhou, China) at 4 °C overnight, followed by incubation with Cy3-conjugated goat anti-rabbit IgG (1:200; Invitrogen, USA) for 1 h. Images were taken by a fluorescence microscope (Olympus, Japan).

Gao W., Liu R., Huang K., Fu W., Wang A., Du G., Tang H., Yin L, & Yin Z.S. (2024). CHMP5 attenuates osteoarthritis via inhibiting chondrocyte apoptosis and extracellular matrix degradation: involvement of NF-κB pathway. Molecular Medicine, 30, 55.

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Molecular Analysis of NP Cell Responses

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After the indicated treatments, NP cells were washed twice with PBS and lysed in radioimmunoprecipitation assay (RIPA) lysis buffer (Beyotime) supplemented with phosphatase and protease inhibitors. The lysates were electrophoresed by 10–12% sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) and transferred onto polyvinylidene difluoride (PVDF) membranes (EMD Millipore, Billerica, MA, USA). Then, the membranes were blocked with NcmBlot blocking buffer (NCM Biotech, Jiangsu, China) for 20 min at room temperature and incubated overnight at 4 °C with primary antibodies against HSP70 (1:1000, ABclonal), Mfn1 (1:1500, Proteintech), Mfn2 (1:1500, Proteintech), OPA1 (1:1000, Proteintech), DRP1 (1:1000, Affinity Biosciences), MFF (1:1000, Proteintech), Fis1 (1:1000, Proteintech), cleaved caspase 3 (1:1000, Affinity Biosciences, OH, USA), caspase 9 (1:1000, Proteintech), Bcl-2 (1:1000, Abcam, Cambridge, MA, USA), Bax (1:1000, Proteintech), Collagen II (1:500, Affinity Biosciences), Aggrecan (1:500, NOVUS, USA), SIRT3 (1:1000, ABclonal), and β-actin (1:2000, Cell Signaling Technology, Danvers, MA, USA). Then, the PVDF membranes were washed with TBST and incubated with horseradish peroxidase-conjugated secondary antibodies for 1 h at room temperature. Signals were detected using an ECL kit (Affinity Biosciences).

Hu B., Wang P., Zhang S., Liu W., Lv X., Shi D., Zhao L., Liu H., Wang B., Chen S, & Shao Z. (2022). HSP70 attenuates compression-induced apoptosis of nucleus pulposus cells by suppressing mitochondrial fission via upregulating the expression of SIRT3. Experimental & Molecular Medicine, 54(3), 309-323.

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