MA3-055

antibody from Invitrogen Antibodies

Targeting: SMARCA5 hISWI, hSNF2H, ISWI

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

MA3-055

Provider

Invitrogen Antibodies

Product name

SMARCA5 Monoclonal Antibody (14SNF-1G2)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

MA3-055 detects SNF2h in human samples. MA3-055 has been succesfully used in western blot application. Immunogen is a synthetic peptide corresponding to residues 1028-1045 of human SNF2h

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

14SNF-1G2

Vial size

50 µL

Concentration

Conc. Not Determined

Storage

-20° C, Avoid Freeze/Thaw Cycles

References

Submitted references

Multiple epigenetic factors co-localize with HMGN proteins in A-compartment chromatin.

He B, Zhu I, Postnikov Y, Furusawa T, Jenkins L, Nanduri R, Bustin M, Landsman D
Epigenetics & chromatin 2022 Jun 27;15(1):23

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of SNF2h was performed by loading 10 µg of HeLa nuclear extract (lane 1), HeLa cytoplasmic extract (lane 2), U2OS nuclear extract (lane 3) and U2OS cytoplasmic extract (lane 4) in reducing sample buffer (Product # 39000) and Page Ruler Plus Protein Ladder (Product # 26619) onto a Novex 4-20% Tris-Glycine polyacrylamide gel (Product # WT4201BX10). Proteins were transferred to nitrocellulose membrane (Product # 88018) with Transfer Buffer (Product # 84731) using the G2 Gast Blotter (Product # 62288). Membrane was blocked in 5% Milk/TBST for one hour at room temperature. SNF2h was detected at approximately 122 kDa using SNF2h monoclonal antibody (Product # MA3-055) at a dilution of 1:2000 overnight at 4°C on a rocking platform, followed by a goat anti-mouse IgG-HRP secondary antibody (Product # 31430) at a dilution of 1:5,000 for one hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34076) and the myECL Imager (Product # 62236).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-SMARCA5 Monoclonal Antibody (14SNF-1G2) (Product # MA3-055) and a 130 kDa band corresponding to SMARCA5 was observed across all tested cell line lysates. Bands marked as * are non-specific bands. Nuclear enriched extracts (30 µg lysate) of HeLa (Lane 1), SH-SY5Y (Lane 2), HEK-293 (Lane 3), Jurkat (Lane 4), K-562 (Lane 5), NIH/3T3 (Lane 6), PC-12 (Lane 7) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23002) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:2000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177,1:10000 dilution) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of SMARCA5 was achieved by transfecting SH-SY5Y with SMARCA5 specific siRNAs (Silencer® select Product # S16081, S16082). Western blot analysis (Fig. A) was performed using Nuclear enriched extracts from the SMARCA5 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with SMARCA5 Monoclonal Antibody (14SNF-1G2) (Product # MA3-055, 1:2000 diution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:10000 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.