MA5-24864

antibody from Invitrogen Antibodies

Targeting: GTF2B TFIIB

Antibody data

Submit

Validation data

Product number: MA5-24864 - Provider product page
Provider: Invitrogen Antibodies
Product name: GTF2B Monoclonal Antibody (3H2)
Antibody type: Monoclonal
Antigen: Recombinant full-length protein
Description: The antibody was affinity-purified from mouse ascites by affinity-chromatography using epitope-specific immunogen and the purity is > 95% (by SDS-PAGE).
Reactivity: Human
Host: Mouse
Isotype: IgG
Antibody clone number: 3H2
Vial size: 100 µL
Concentration: 1 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

GTF2B protein structure - MA5-24864 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of GTF2B in Jurkat, MCF7 and HeLa cell lysates. Samples were incubated with GTF2B monoclonal antibody (Product # MA5-24864) at a dilution of 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of GTF2B was achieved by transfecting HeLa cells with GTF2B specific siRNAs (Silencer® select Product # s6295, s6296). Western blot analysis (Fig. a) was performed using modified whole cell extracts (1% SDS) from the GTF2B knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blots were probed with Anti-GTF2B Monoclonal Antibody (3H2) (Product # MA5-24864, 1:1000 dilution) and Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 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 GTF2B. An uncharacterized band around 60 kDa was also observed.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-GTF2B Monoclonal Antibody (3H2) (Product # MA5-24864) and 36 kDa band corresponding to GTF2B was observed across the cell lines tested. An uncharacterized band of ~ 60 kDa was also observed in all the cell lines. Modified whole cell extracts (1% SDS) (30 µg lysate) of HeLa (Lane1), K-562 (Lane 2), MOLT-4 (Lane 3), Hep G2 (Lane 4) and Jurkat (Lane 5) were electrophoresed using Novex® NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with 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

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of GTF2B in HeLa cells. Samples were incubated with GTF2B monoclonal antibody (Product # MA5-24864) at a dilution of 1:50.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of DLAT in HeLa cells. Samples were incubated with DLAT monoclonal antibody (Product # MA5-24863) at a dilution of 1:300.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of GTF2B in HeLa cells. Samples were incubated with GTF2B monoclonal antibody (Product # MA5-24864) at a dilution of 1:50.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of GTF2B in HeLa cells. Samples were incubated with GTF2B monoclonal antibody (Product # MA5-24864) at a dilution of 1:50.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of GTF2B was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with GTF2B Monoclonal Antibody (3H2) (Product # MA5-24864) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then with Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32731) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.