PA1-25450

antibody from Invitrogen Antibodies

Targeting: SMARCB1 BAF47, hSNFS, Ini1, PPP1R144, RDT, Sfh1p, SNF5, SNF5L1, Snr1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA1-25450

Provider

Invitrogen Antibodies

Product name

SMARCB1 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

Excitation: 405 nm; Emission: 436 nm; Laser: Violet Laser

Reactivity

Human, Mouse

Host

Rabbit

Isotype

IgG

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.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of SMARCB1 was achieved by transfecting HeLa cells with SMARCB1 specific siRNAs (Silencer® select Product # s529255, s13142). Western blot analysis (Fig. a) was performed using 20ug of modified whole cell extracts (1% SDS) from the HeLa knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with SMACRB1 Rabbit Polyclonal Antibody (Product # PA1-25450, 0.25 µg/mL) and Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 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 SMARCB1.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-SMARCB1 Polyclonal Antibody (Product # PA1-25450) and a ~44 kDa band was observed across the panel tested. Modified Whole cell extracts (1% SDS) (30 µg lysate) of A549 (Lane 1), PC-3 (Lane 2), HeLa (Lane 3), K-562 (Lane 4), HEK-293 (Lane 5), MCF-7 (Lane 6), U-2 OS (Lane 7) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). 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 µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of SMARCB1 was performed using 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 SMARCB1 Polyclonal Antibody (Product # PA1-25450) at 1:200 dilution in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 (Product # A27034, 1:2000 dilution) for 45 minutes at room temperature (Panel a: green) in HeLa cells. Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image of HeLa cells showing nuclear localization for SMARCB1. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.