MA5-15638

antibody from Invitrogen Antibodies

Targeting: GATA1 ERYF1, GATA-1, GF1, NF-E1, NFE1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Chromatin Immunoprecipitation (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-15638

Provider

Invitrogen Antibodies

Product name

GATA1 Monoclonal Antibody (4F5)

Antibody type

Monoclonal

Antigen

Purifed from natural sources

Description

MA5-15638 targets GATA1 in IF, IHC, and WB applications and shows reactivity with Human samples.

Antibody clone number

4F5

Concentration

Conc. Not Determined

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of GATA1 using GATA1 monoclonal antibody (Product # MA5-15638) in K562 (1) cell lysate.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of GATA1 using GATA1 monoclonal antibody (Product # MA5-15638) in K562 (1) cell lysate.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using GATA1 Mouse Monoclonal Antibody (Product # MA5-15638) and a 43kDa band corresponding to GATA1 was observed across cell lines except Jurkat and A-431 along with an uncharacterized band (*) at ~52 kDa in the cell lines tested. Whole cell extracts (30 µg lysate) of K-562 (Lane 1), HEL 92.1.7 (Lane 2), Jurkat (Lane 3) and A-431 (Lane 4) were electrophoresed using 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).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of K562 (left) cells using GATA1 monoclonal antibody (Product # MA5-15638) (Green). Blue: DRAQ5 fluorescent DNA dye.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of GATA1 was performed using log phase K-562 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 GATA1 Monoclonal Antibody (4F5) (Product # MA5-15638) at 1:200 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, Alexa Fluor 488 conjugate (Product # A28175) 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 staining in nucleus. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of paraffin-embedded pancreatic cancer, using GATA1 monoclonal antibody (Product # MA5-15638) followed with DAB staining.

Chromatin Immunoprecipitation

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Chromatin Immunoprecipitation (ChIP) assay of endogenous GATA1 protein using Anti-GATA1 Antibody: ChIP was performed using Anti-GATA1 Mouse Monoclonal Antibody (Product # MA5-15638, 5 µg) on sheared chromatin from K562 cells using the MAGnify ChIP System kit (Product # 49-2024). Normal Mouse IgG was used as a negative IP control. The purified DNA was analyzed by qPCR using primers binding to FCER1A promoter and SAT2 satellite repeats. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.