MA5-23508

antibody from Invitrogen Antibodies

Targeting: SMARCA5 hISWI, hSNF2H, ISWI

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Data presented on provider website)

Immunoprecipitation (Recommended by provider)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-23508

Provider

Invitrogen Antibodies

Product name

Anti-SMARCA5 Monoclonal Antibody

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Reactivity

Human

Host

Mouse

Isotype

IgG

Vial size

100 µl

Concentration

Lot Dependent

Storage

-20°C or -80°C if preferred

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot was performed on nuclear extracts from HeLa cells (HeLa NE, 20 µg) using a SNF2H/SMARCA5 monoclonal antibody (Product # MA5-23508) at a dilution of 1:1,000 in TBS-Tween containing 5% skimmed milk. The molecular weight marker (kDa) is shown on the left), the position of the protein of interest is shown on the right.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of SMARCA5 was achieved by transfecting HeLa with SMARCA5 specific siRNAs (Silencer® select Product # s16081, s16082). Western blot analysis (Fig. a) was performed using modified whole cell extracts (1% SDS) from the SMARCA5 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with SMARCA5 Monoclonal Antibody (Product # MA5-23508, 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). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to SMARCA5.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-SMARCA5 Monoclonal Antibody (Product # MA5-23508) and a 130kDa band corresponding to SMARCA5 was observed across all cell lines tested. Modified whole cell extracts (30 µg lysate) of K-562 (Lane 1), HeLa (Lane 2), Raji (Lane 3), U-2 OS (Lane 4), Hep G2 (Lane 5) and MOLT-4 (Lane 6) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris 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 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)..

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of SMARCA5 Monoclonal Antibody 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 10 minutes, and blocked with 2% BSA for 10 minutes at room temperature. The cells were labeled with SMARCA5 Monoclonal Antibody (Product # MA5-23508) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175, 1:2000 dilution) 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..