- MA5-26235 - Invitrogen Antibodies TRIM38 antibody

Lu Z, Deng M, Ma G, Chen L
PeerJ 2022;10:e13815

Lu Z, Hao C, Qian H, Zhao Y, Bo X, Yao Y, Ma G, Chen L
iScience 2022 Aug 19;25(8):104780

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of TRIM38 in HepG2, HeLa, SVT2, A549, COS7, Jurkat, MDCK, PC12 and MCF7 cells using 35 µg per lane. Samples were separated by SDS-PAGE and probed with TRIM38 (Product # MA5-26235) monoclonal antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of TRIM38 in COS7 cells. Cells were transfected with a plasmid overexpressing TRIM38 and probed with a TRIM38 monoclonal antibody (Product # MA5-26235).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry was performed on paraffin-embedded human endometrium tissue. To expose target proteins, 10mM citric buffer, pH6.0, 120°C for 3min was used. Following antigen retrieval, tissues were probed with a TRIM38 monoclonal antibody (Product # MA5-26235).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry was performed on paraffin-embedded adenocarcinoma of human ovary tissue. To expose target proteins, 10mM citric buffer, pH6.0, 120°C for 3min was used. Following antigen retrieval, tissues were probed with a TRIM38 monoclonal antibody (Product # MA5-26235).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on paraffin-embedded carcinoma of human thyroid tissue. To expose target proteins, 10mM citric buffer, pH6.0, 120°C for 3min was used. Following antigen retrieval, tissues were probed with a TRIM38 monoclonal antibody (Product # MA5-26235).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: 10.7717/peerj.13815/fig-1 Figure 1 TRIM38 expression and its roles in H9c2 cells exposed to H/R. H9c2 cells were exposed to 6 h of hypoxia, followed by 6 h of reoxygenation. (A) Representative immunoblotting analysis and quantification of TRIM38, Bax, Bcl-2, C-caspase3, and caspase3 protein levels in H9c2 cells insulted by H/R or not. (B) Calculation of the ratio of Bax/Bcl2 and C-caspase3/caspase3. n = 3 for each group. ** p < 0.01 and *** p < 0.001 versus control. Unpaired two-tailed student t test for A and B.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: 10.7717/peerj.13815/fig-2 Figure 2 TRIM38 attenuates apoptosis and oxidative stress in H/R model. (A) Representative immunoblotting analysis (Left) and quantification (right) of TRIM38 expression in cultured H9c2 cells transfected with AdGFP or AdTRIM38. n = 4 for each group. (B) Representative immunoblotting analysis (Left) and quantification (right) of Bax, Bcl-2, C-caspase3, and caspase3 protein levels in H9c2 cells transfected with AdGFP or AdTRIM38 after H/R or not. (C) Calculation of the ratio of Bax/Bcl2 and C-caspase3/caspase3 in Control or H/R-treated H9c2 cells. (D) Effects of TRIM38 overexpression on LDH release in H/Rmodel. (E) Cellular MDA content in H/R model. (F) Cellular SOD activities in H/R model. (G) Cellular GSH-Px activities in H/R model. n = 3 for each group. * p < 0.05, ** p < 0.01 and *** p < 0.001. Unpaired two-tailed student t test for A. One-way ANOVA followed by post-hoc tests for B-G.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: 10.7717/peerj.13815/fig-3 Figure 3 TRIM38 knockdown enhanced apoptosis in H9c2 Cells After H/R. (A) Representative immunoblotting analysis (Left) and quantification (right) of TRIM38 expression in cultured H9c2 cells transfected with AdshRNA or AdshTRIM38. (B) Representative immunoblotting analysis (Left) and quantification (right) of Bax, Bcl-2, C-caspase3, and caspase3 protein levels in H9c2 cells transfected with AdshRNA or AdshTRIM38 after H/R or not. (C) Calculation of the ratio of Bax/Bcl2 and C-caspase3/caspase3 in Control or H/R-treated H9c2 cells. n = 3 for each group. * p < 0.05, ** p < 0.01 and *** p < 0.001. One-way ANOVA followed by post-hoc tests for A-C.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 10.7717/peerj.13815/fig-6 Figure 6 Under H/R conditions, TRIM38 degraded TRAF6 viaubiquitination. (A) H9c2 cells were subjected to H/R stimulation. Cell lysate and normal IgG antibody or TRAF6 antibody were subjected to Co-IP, and the indicated antibodies were used for immunoblotting analysis. (B-C) TRIM38 knockdown or overexpression cells were exposed with H/R. Immunoprecipitation assay was performed using lysates with anti-TRAF6 antibody. IB assay was performed using anti-TRAF6, anti-TRIM38, and anti-Ub antibodies. (D) Working model of TRIM38 protecting H9c2 cells from H/R injury by inhibiting TRAF6/TAK1/NF- kappa B signaling pathway. TRIM38 alleviated H/R-induced H9c2 injury by promoting TRAF6 degradation, which led to the inactivation of TAK1, an upstream NF- kappa B signalling molecule.

MA5-26235