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Purified anti-NFATc1 Antibody

Pricing & Availability
Clone
7A6 (See other available formats)
Regulatory Status
RUO
Other Names
Nuclear factor of activated T-cells, cytoplasmic 1, NFAT transcription complex cytosolic component, NFAT2, NFATc
Isotype
Mouse IgG1, κ
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Product Citations
publications
1_7A6_PURE_NFATc1_102017
Total lysates (15 µg protein) from EL4 (lane 1), HeLa (lane 2), Jurkat (lane 3) and Raw 264.7 cells (lane 4) were resolved by electrophoresis (4-20% Tris-Glycine gel), transferred to nitrocellulose, and probed with 1:500 diluted (1 µg/mL) Purified anti-NFATc1 Antibody, clone 7A6 (upper). Proteins were visualized by chemiluminescence detection using a 1:3000 diluted goat anti-mouse-IgG secondary antibody conjugated to HRP for the anti-NFATc1 Antibody or 1:5000 diluted Direct-Blot HRP anti-β-Actin Antibody, clone 2F1-1(lower). Lane M: Molecular weight ladder.
  • 1_7A6_PURE_NFATc1_102017
    Total lysates (15 µg protein) from EL4 (lane 1), HeLa (lane 2), Jurkat (lane 3) and Raw 264.7 cells (lane 4) were resolved by electrophoresis (4-20% Tris-Glycine gel), transferred to nitrocellulose, and probed with 1:500 diluted (1 µg/mL) Purified anti-NFATc1 Antibody, clone 7A6 (upper). Proteins were visualized by chemiluminescence detection using a 1:3000 diluted goat anti-mouse-IgG secondary antibody conjugated to HRP for the anti-NFATc1 Antibody or 1:5000 diluted Direct-Blot HRP anti-β-Actin Antibody, clone 2F1-1(lower). Lane M: Molecular weight ladder.
  • 2_7A6_GoChIP_NFATc1_Antibody_1_120717.png
    Chromatin Immunoprecipitation (ChIP) was performed using Go-ChIP-Grade™ Protein G Enzymatic kit by loading 3 µg of cross-linked chromatin samples from Raw264.7 cells treated with RANKL with either A) 1:500 dilution of Go-ChIP-Grade™ Purified anti-NFATc1 (Clone 7A6), or B) equal amount of Purified Mouse IgG1 Isotype Control Antibody. The enriched DNA was purified and quantified by real-time qPCR using primers targeting mouse TRAP gene region. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the 5% of total amount of input chromatin.
  • 7A6_GoChIP_NFATc1_Antibody_2_120717.png
    Chromatin Immunoprecipitation (ChIP) was performed using Go-ChIP-Grade™ Protein G Enzymatic kit by loading 3 µg of cross-linked chromatin samples from Raw264.7 cells treated with RANKL with either A) 1:500 dilution of Go-ChIP-Grade™ Purified anti-NFATc1 (Clone 7A6), or B) equal amount of Purified Mouse IgG1 Isotype Control Antibody. The enriched DNA was purified and quantified by real-time qPCR using primers targeting mouse OSCAR gene region. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the 5% of total amount of input chromatin.
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649601 25 µg $124
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649602 100 µg $268
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Description

The product of this gene is a component of the nuclear factor of activated T cells DNA-binding transcription complex. The protein complex consists of NFAT1, NFAT2 (NFATc1 or NFATc), NFAT3, and NFAT4. All members of this family are transcription factors with a Rel homology domain and regulate gene transcription in concert with AP-1 (Jun/Fos) to orchestrate an effective immune response. NFAT proteins are predominantly expressed in cells of the immune system but are also expressed in skeletal muscle, keratinocytes and adipocytes, regulating cell differentiation programs in these cells. In resting cells, NFAT proteins are heavily phosphorylated and localized in the cytoplasm. Increased intracellular calcium concentrations activate the calcium/calmodulin-dependent serine phosphatase calcineurin, which dephosphorylates NFAT proteins, resulting in their subsequent translocation to the nucleus.

Proteins belonging to this family of transcription factors play a central role in inducible gene transcription during immune response. The product of this gene is an inducible nuclear component. It functions as a major molecular target for the immunosuppressive drugs such as cyclosporin A. Five transcript variants encoding distinct isoforms have been identified for this gene. Different isoforms of this protein may regulate inducible expression of different cytokine genes.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Human, Mouse, Rat
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Recombinant protein of human NFATc1 amino acids 197-304.
Formulation
This antibody is provided in phosphate-buffered solution, pH 7.2, 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
Upon receipt, store undiluted between 2°C and 8°C.
Application

WB - Quality tested
ChIP - Verified
FC - Reported in the literature, not verified in house
ICC - Reported by the developer, not verified in house
Recommended Usage

Each lot of this antibody is quality control tested by Western blotting. For Western blotting applications, a concentration of 1 µg/mL is recommended. For ChIP applications, the suggested dilution is is 1:100-1:2000 by volume. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

7A6 antibody detects endogenous human NFATC1 in Western blot. There are 10 isoforms with a predicted MW of 39kD, 74kD, 76kD, 77kD (3), 88kD (2), 100kD, 101kD. With this antibody, observed MW bands range from 70 - 120 kD. The optimal concentration should be determined by titration for each individual assay of interest.

Clone 7A6 Intracellular Staining Procedure:

  1. Prepare the fix/perm solution, consisting of 1% paraformaldehyde in 70% ethanol. Chill for at least one hour at -20°C.
  2. Suspend cells in cell staining buffer and perform surface stain if necessary.
  3. Centrifuge for 5 minutes at 1500 RPM and discard supernatant.
  4. Resuspend cells in 500 µL of fix/perm solution for up to 2 x 10^6 cells and gently vortex.
  5. Incubate cells on ice for 30-60 minutes in the dark.
  6. Add 2 mL PBS and centrifuge cells for 1500 RPM for 5 minutes.
  7. Wash cells 1X with 2 mL cell staining buffer, centrifuge at 1500 RPM for 5 minutes, discard supernatant.
  8. Adjust cell concentration to about 1 x 10^7 cells/mL, add 100 µL (~ 1 x 10^6 cells) to a FACS tube that already contains the staining antibody and mix gently.
  9. Incubate 15 minutes at room temperature.
  10. Wash 2X with cell staining buffer as in step 6.
  11. Resuspend cells in cell staining buffer and analyze.
Application References

(PubMed link indicates BioLegend citation)
  1. Timmerman LA, et al. 1997. J. Immunol. 159:2735. (IF)
  2. Brandt C, et al. 2010. Cytometry A. 77:607. (FC)
  3. Fan W, et al. 2012. Arthritis Rheum. 64:3715. PubMed
Product Citations
  1. Flora P, et al. 2021. PLoS Genet. 17:e1009948. PubMed
  2. Kresnoadi U, et al. 2021. Saudi Dent J. 33:1055. PubMed
  3. Liu F, et al. 2020. J Immunother Cancer. 8:. PubMed
  4. Brummer G, et al. 2020. Oncogene. 39:2275. PubMed
  5. Li X, et al. 2019. Cell Death Discov. 5:62. PubMed
  6. Abkhezr M, et al. 2015. Am J Physiol Renal Physiol. 309: F98 - F108. PubMed
  7. Murata K et al. 2017. Immunity. 47(1):66-79 . PubMed
  8. Fan W, et al. 2012. Arthritis Rheum. 64:3715. PubMed
RRID
AB_10680239 (BioLegend Cat. No. 649601)
AB_10680239 (BioLegend Cat. No. 649602)

Antigen Details

Structure
943 amino acids with predicted molecular weight of 101kD. There are 10 isoforms with a predicted MW of 39kD, 74kD, 76kD, 77kD (3), 88kD (2), 100kD, 101kD.
Distribution

Cytoplasm, Nucleus. NFATc1 is cytoplasmic in the phosphorylated form and nuclear after calcineurin-mediated dephosphorylation controlled activation.

Biology Area
Cell Biology, Immunology, Neuroscience, Neuroscience Cell Markers, Signal Transduction, Transcription Factors
Molecular Family
Nuclear Markers
Antigen References

1. Zhao Q, et al. 2010. Int. J. Biochem. Cell Biol. 42:576.
2. Hoey T, et al. 1995. Immunity 2:461.
3. Northrop JP, et al. 1993. J. Biol. Chem. 268:2917.
4. Hogan PG, et al. 2003. Genes Dev. 17:2205.
5. Shaw KT, et al. 1995. P. Natl. Acad. Sci. USA 92:11205.
6. Serfling E, et al. 2007. Sci. STKE 138:pe42.

Gene ID
4772 View all products for this Gene ID
UniProt
View information about NFATc1 on UniProt.org
Go To Top Version: 7    Revision Date: 06/15/2022

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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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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