MA5-12736

antibody from Invitrogen Antibodies

Targeting: TGM2 TGC

Immunoprecipitation

Antibody data

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Validation data

TGM2 protein structure - MA5-12736 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot of Transglutaminase II using Transglutaminase II Monoclonal Antibody (Product # MA5-12736) on HUVEC Cells.
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of HeLa (Lane 1), MDA-MB-231 (Lane 2), K-562 (Lane 3) and A549 (Lane 4). The blots were probed with Mouse anti-TGM2 Monoclonal Antibody (Product # MA5-12736, 2 µg/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). A 78 kDa band corresponding to TGM2 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), 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 antibody following blocking with 5 % skimmed milk. This is followed by incubating the membrane with Poly-HRP Streptavidin (Product # N200, 1:10,000). Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of TGM2 was achieved by transfecting HeLa cells with TGM2 specific siRNAs (Silencer® select Product # s14087). Western blot analysis (Fig a) was performed using Membrane extract from the TGM2 knock down cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Anti-TGM2 biotin Mouse Monoclonal Antibody (Product # MA5-12736, 0.2 µg/mL) and Poly-HRP Streptavidin (Product # N200, 1:10,000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig b). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to TGM2.
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockout of TGM2 was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR979226_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of TGM2 was performed by loading 30 µg of HeLa Wild type (Lane 1), HeLa Cas9 (Lane 2) and HeLa TGM2 KO (Lane 3) whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-TGM2 Monoclonal Antibody (CUB 7402), Biotin (Product # MA5-12736, 0.2 µg dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:5,000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to TGM2.
Conjugate: Biotin