MA5-14238

antibody from Invitrogen Antibodies

Targeting: MMP13 CLG3

Immunoprecipitation

Antibody data

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Validation data

MMP13 protein structure - MA5-14238 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of PC-3 (Lane 1), COLO205 (Lane 2), HeLa (Lane 3) and MDA-MB-231 (Lane 4). The blots were probed with Anti-MMP-13 Mouse Monoclonal Antibody (Product # MA5-14238, 2-4 µg/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Secondary Antibody, HRP conjugate (Product # 62-6520, 1:4000 dilution). A 53 kDa band corresponding to MMP-13 was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0301BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of PC-3 (Lane 1), COLO205 (Lane 2), HeLa (Lane 3) and MDA-MB-231 (Lane 4). The blots were probed with Anti-MMP-13 Mouse Monoclonal Antibody (Product # MA5-14238, 2-4 µg/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Secondary Antibody, HRP conjugate (Product # 62-6520, 1:4000 dilution). A 53 kDa band corresponding to MMP-13 was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0301BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-MMP13 Monoclonal Antibody (VIIIA2) (Product # MA5-14238) and a 50 kDa band corresponding to the activated form of MMP-13 was observed in all the tested cell lines. Whole cell lysates (30 µg lysate) of HeLa (Lane 1), COLO 205 (Lane 2), MDA-MB-231 (Lane 3), Jurkat (Lane 4), PC-3 (Lane 5), U-2 OS (Lane 6) and Hep G2 (Lane 7) were electrophoresed using NuPAGE® 10 % Bis-Tris gel (Product # NP0302BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blots were probed with the primary antibody (3 µg/mL) and detected by chemiluminescence Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5 TAP2 efficiently suppresses the expression of proinflammatory cytokines and MMP13. ( a ) Primary human chondrocytes were incubated with TAP2 (10 muM, 3 h) prior to LPS treatment (0.1 ng/mL, 3 h) or ( b ) primary human synoviocytes were incubated with TAP2 (100 muM, 3 h) prior to LPS treatment (100 ng/mL, 3 h). Then, total RNA was isolated from the cells and utilized for the analysis of the inflammatory mediators, such as TNF-alpha, IL-1beta, IL-6, MMP3, or MMP13, in chondrocytes and TNF-alpha, IL-1beta, and IL-6 in synoviocytes by quantitative PCR. The data are expressed as the mean +- SEM (one-way ANOVA test, **** P < 0.0001, *** P < 0.001, ** P < 0.01, * P < 0.05 vs. LPS + TAP2). ( c ) On day 3 post-injection of PBS and TAP2, the knee joint tissues of MIA-induced arthritic rats (n = 3) were incubated with DHE solution (1 muM) to visualize ROS or immunostained with an anti-MMP13 antibody. The intensity of ROS attaining or MMP13 was quantified in cartilages using ImageJ. The data are expressed as the mean +- SEM (one-way ANOVA test, **** P < 0.0001, *** P < 0.001 vs. MIA). ( d ) Total RNA from the knee joint cartilage of MIA-induced arthritic rats, MIA arthritis rats treated with TAP2, and control animals was isolated and analyzed by quantitative PCR (n = 3 per each group). The data are expressed as the mean +- SEM (one-way ANOVA test, ** P < 0.01, * P < 0.05 vs. MIA).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig 6 Expression of MMP-13 in human lung adenocarcinoma. Immunohistochemical staining was performed using Clone VIIIA2 diluted 1:20 (Lab Vision Corp., CA). A) Solid predominant adenocarcinoma. MMP-13 index staining (brown) is high in tumor cells (black arrow) and in fibroblasts (yallow arrow) (x100). B) Minimally invasive adenocarcinoma. MMP-13 staining is high in tumor cells (black arrow), and high in fibroblasts (yellow arrow) (x100). C) Adenocarcinoma in-situ . MMP-13 staining is high in tumor cells (black arrow), and high in fibroblasts (yellow arrow) (x100). D) Solid predominant adenocarcinoma and normal lung. Note the absence of MMP-13 staining in normal lung (*). Scale bar: 500 mum. Inserts in the x100 IHCs show x400 magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 4. Immunohistochemical analysis results for MMP-13 in the in vivo model of OA. MMP-13 staining was determined by IHC in in tissue samples from mice in the Sham, OA and OA + Res groups. Strong staining was considered to indicate a MMP-13-positive Result. (Scale bar=10 um). MMP-13, matrix metallopeptidase 13; OA, osteoarthritis; Res, Resveratrol; IHC, immunohistochemistry.