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Purified anti-GATA1 Antibody

Pricing & Availability
Clone
P84F5 (See other available formats)
Regulatory Status
RUO
Other Names
GATA Binding Protein 1, Erythroid Transcription Factor, Globin Transcription Factor 1, NFE1, XLTT
Isotype
Mouse IgG2b, κ
Ave. Rating
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Product Citations
publications
P84F5_PURE_GATA1_Antibody_1_081820
Whole cell extracts (15 µg total protein) from K-562 and HEL (positive controls), and HEK-293 and HeLa (negative controls) cells were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 0.25 µg/mL (1:2000 dilution) of purified anti-GATA1 antibody (clone P84F5) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG antibody (Cat. No. 405306) at a 1:3000 dilution. Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 607904) was used as a loading control at a 1:25000 dilution (lower). Lane M: Molecular weight marker.
  • P84F5_PURE_GATA1_Antibody_1_081820
    Whole cell extracts (15 µg total protein) from K-562 and HEL (positive controls), and HEK-293 and HeLa (negative controls) cells were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 0.25 µg/mL (1:2000 dilution) of purified anti-GATA1 antibody (clone P84F5) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG antibody (Cat. No. 405306) at a 1:3000 dilution. Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 607904) was used as a loading control at a 1:25000 dilution (lower). Lane M: Molecular weight marker.
  • P84F5_PURE_GATA1_Antibody_2_081820
    HeLa (negative control, panel A) and HEL (positive control, panel B) cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.5% Triton X-100 for 10 minutes, and blocked with 5% FBS for 60 minutes. Cells were then intracellularly stained with 2.0 µg/mL (1:250 dilution) of purified anti-GATA1 antibody (clone P84F5) overnight at 4°C, followed by incubation with Alexa Fluor® 594 goat anti-mouse IgG antibody (Cat. No. 405326) at 2.0 µg/mL. Nuclei were counterstained with DAPI, and the image was captured with a 60X objective.
  • P84F5_PURE_GATA1_Antibody_3_081820
    Whole cell extracts (250 µg total protein) prepared from HEL cells were immunoprecipitated overnight with 2.5 µg of purified mouse IgG2b, κ isotype ctrl antibody (Cat. No. 400302) or purified anti-GATA1 antibody (clone P84F5). The resulting IP fractions, whole cell extract input (6%), and post-IP supernatants (Sup) were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with clone P84F5. Lane M: Molecular weight marker.
  • P84F5_PURE_GATA1_Antibody_4_012921
    HEL cells (filled histogram, positive control) or HeLa cells (open histogram, negative control) were fixed and permeabilized using the True-Nuclear™ Transcription Factor Buffer set (Cat. No. 424401), and intracellularly stained with purified anti-GATA1 antibody (clone P84F5) followed by PE goat anti-mouse IgG antibody (Cat. No. 405307).
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939101 25 µg 136 CHF
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939102 100 µg 335 CHF
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Description

The GATA family of transcription factors is comprised of six members characterized by their binding to the consensus DNA sequence (T/A)GATA(A/G). GATA1, which was the initially described member of the GATA family, is predominantly expressed in cells of the myeloid lineage such as erythrocytes, eosinophils, mast cells, and megacaryocytes. Over the course of erythroid differentiation, GATA2 activates expression of the functionally distinct GATA1; promoter-occupied GATA2 is replaced/exchanged at certain promoters with the GATA1 transcription factor, a process known as the GATA switch. Genes induced by GATA1 include HBB, HBG1/2, ALAS2 and HMBS. Haploinsufficient heterozygous GATA1 knockout mice spontaneously develop erythroblastic leukemia, and GATA1-null mice die early in development due to profound anemia.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Full-length recombinant human GATA1 protein
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
ICC, IP, ICFC - Verified
Recommended Usage

Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested use of this reagent is 0.1 - 1.0 µg/mL. For immunocytochemistry, a concentration range of 1.0 - 5.0 μg/mL is recommended. For immunoprecipitation, the suggested use of this reagent is 2.5 µg/test. For intracellular flow cytometric staining, the suggested use of this reagent is ≤ 0.5 µg per million cells in 100 µL volume. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

This clone was tested for ICC using 4% PFA-fixed HEL cells permeabilized with either methanol or Triton X-100. While both methods were compatible with robust HEL staining, Triton X-100 permabilization resulted in stronger staining.

When using this clone for ICFC, we recommend using the True-Nuclear™ Transcription Factor Buffer Set or the True Phos perm buffer. We do not recommend using the Intracellular Staining Permeabilization Wash Buffer for ICFC testing due to poor GATA1 staining.

True-Nuclear™ Transcription Factor Buffer Set allowed for better binding to the epitope when compared to the True Phos perm buffer, there was a twofold difference between their median PE readings.

RRID
AB_2876765 (BioLegend Cat. No. 939101)
AB_2876765 (BioLegend Cat. No. 939102)

Antigen Details

Structure
GATA1 is a 413 amino acid protein with a predicted molecular weight of 43 kD. Additional isoforms of 335 (35 kD) and 330 (34 kD) amino acids have been described.
Distribution

Erythroid and megakaryocytic cells/Nucleus

Function
Transcription Factor/Erythroid cell development and differentiation
Cell Type
Erythrocytes, Megakaryocytes
Biology Area
Cell Biology, Stem Cells, Transcription Factors
Antigen References
  1. Shimizu R, et al. 2008. Nat Rev Cancer. 8:279-87.
  2. Bresnick EH, et al. 2010. J Biol Chem. 285:31087-93.
  3. Katsumura, et al. 2017. Blood. 129:2092-2102.
Gene ID
2623 View all products for this Gene ID
UniProt
View information about GATA1 on UniProt.org
Go To Top Version: 2    Revision Date: 01.29.2021

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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