702414

antibody from Invitrogen Antibodies

Targeting: ACTL6A Actl6, Arp4, BAF53A, INO80K

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

702414

Provider

Invitrogen Antibodies

Product name

BAF53A Recombinant Rabbit Monoclonal Antibody (5H3L6)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

This antibody is predicted to react with Monkey, Pig, Rat

Antibody clone number

5H3L6

Concentration

0.5 mg/mL

References

Submitted references

Increased ACTL6A occupancy within mSWI/SNF chromatin remodelers drives human squamous cell carcinoma.

Chang CY, Shipony Z, Lin SG, Kuo A, Xiong X, Loh KM, Greenleaf WJ, Crabtree GR
Molecular cell 2021 Dec 16;81(24):4964-4978.e8

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on Nuclear enriched extracts (30 µg lysate) of MCF-7 (Lane 1), K-562 (Lane 2), C2C12 (Lane 3), Jurkat (Lane 4), Ramos (Lane 5) and tissue extracts of Mouse Testis (Lane 6) and Rat Testis (Lane 7). The blots were probed with Anti-BAF53A Recombinant Rabbit Monoclonal Antibody (Product # 702414, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 47 kDa band corresponding to BAF53A was observed across the cell lines and tissue 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® 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).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

For immunofluorescence analysis, MDA-MB-231 cells were fixed and permeabilized for detection of endogenous BAF53A using Anti-BAF53A recombinant rabbit monoclonall Antibody (Product # 702414, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of BAF53A protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating nuclear localization of BAF53A. Panel e)represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.