Overview

  • Product name

    Anti-NFAT2 antibody [7A6] - ChIP Grade
    See all NFAT2 primary antibodies
  • Description

    Mouse monoclonal [7A6] to NFAT2 - ChIP Grade
  • Host species

    Mouse
  • Tested applications

    Suitable for: ChIP, IHC-FoFr, ICC/IF, Flow Cyt, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Hamster, Human, Non human primates
  • Immunogen

    Fusion protein corresponding to NFAT2 aa 1-700.

  • Positive control

    • IHC-P: FFPE human Hodgkin's lymphoma tissue sections.
  • General notes

    This monoclonal antibody is manufactured by Abcam. If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find more information here.

Properties

Applications

Our Abpromise guarantee covers the use of ab2796 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ChIP Use at an assay dependent concentration. PubMed: 20375015
IHC-FoFr Use at an assay dependent concentration. PubMed: 20530871
ICC/IF 1/250.
EMSA Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

IHC-P Use at an assay dependent concentration. PubMed: 16947118

Target

  • Function

    Plays a role in the inducible expression of cytokine genes in T-cells, especially in the induction of the IL-2 or IL-4 gene transcription. Also controls gene expression in embryonic cardiac cells. Could regulate not only the activation and proliferation but also the differentiation and programmed death of T-lymphocytes as well as lymphoid and non-lymphoid cells.
  • Tissue specificity

    Expressed in thymus, peripheral leukocytes as T-cells and spleen. Isoforms A are preferentially expressed in effector T-cells (thymus and peripheral leukocytes) whereas isoforms B and isoforms C are preferentially expressed in naive T-cells (spleen). Isoforms B are expressed in naive T-cells after first antigen exposure and isoforms A are expressed in effector T-cells after second antigen exposure.
  • Sequence similarities

    Contains 1 RHD (Rel-like) domain.
  • Domain

    Rel Similarity Domain (RSD) allows DNA-binding and cooperative interactions with AP1 factors.
    The N-terminal transactivation domain (TAD-A) binds to and is activated by Cbp/p300. The dephosphorylated form contains two unmasked nuclear localization signals (NLS), which allow translocation of the protein to the nucleus.
    Isoforms C have a C-terminal part with an additional trans-activation domain, TAD-B, which acts as a transcriptional activator. Isoforms B have a shorter C-terminal part without complete TAD-B which acts as a transcriptional repressor.
  • Post-translational
    modifications

    Phosphorylated by NFATC-kinase; dephosphorylated by calcineurin.
  • Cellular localization

    Cytoplasm. Nucleus. Cytoplasmic for the phosphorylated form and nuclear after activation that is controlled by calcineurin-mediated dephosphorylation. Rapid nuclear exit of NFATC is thought to be one mechanism by which cells distinguish between sustained and transient calcium signals. The subcellular localization of NFATC plays a key role in the regulation of gene transcription.
  • Information by UniProt
  • Database links

  • Alternative names

    • cytoplasmic 1 antibody
    • MGC138448 antibody
    • NF ATc antibody
    • NF ATc1 antibody
    • NF-ATc antibody
    • NF-ATc1 antibody
    • NF-ATc1.2 antibody
    • NFAC1_HUMAN antibody
    • NFAT 2 antibody
    • NFAT transcription complex cytosolic component antibody
    • NFATC 1 antibody
    • NFATc antibody
    • NFATc1 antibody
    • Nuclear factor of activated T cells cytoplasmic 1 antibody
    • Nuclear factor of activated T cells cytoplasmic calcineurin dependent 1 antibody
    • Nuclear factor of activated T cells cytosolic component 1 antibody
    • nuclear factor of activated T-cells 'c' antibody
    • Nuclear factor of activated T-cells antibody
    see all

Images

  • IHC image of NFAT2 staining in a section of formalin-fixed paraffin-embedded normal human Hodgkin's lymphoma* performed on a Leica BONDTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab2796, 1ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • ab2796 staining human normal tonsil tissue. Staining is localized to cytoplasm and nucleus.
    Left panel: with primary antibody at 1 µg/ml. Right panel: Isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS), then incubated with primary antibody for 20 minutes, and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with hematoxylin and coverslipped under DePeX.

    Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

  • Overlay histogram showing Jurkat cells stained with ab2796 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab2796, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

References

This product has been referenced in:

  • Soares MPR  et al. The use of apocynin inhibits osteoclastogenesis. Cell Biol Int N/A:N/A (2019). Read more (PubMed: 30761659) »
  • Oh-Hora M  et al. Stromal Interaction Molecule Deficiency in T Cells Promotes Spontaneous Follicular Helper T Cell Development and Causes Type 2 Immune Disorders. J Immunol 202:2616-2627 (2019). Read more (PubMed: 30910863) »
See all 26 Publications for this product

Customer reviews and Q&As

1-10 of 13 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Liver)
Permeabilization
Yes - 0.1%Triton x-100 in PB
Specification
Liver
Blocking step
Serum as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 10% · Temperature: 4°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Jul 16 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Liver)
Antigen retrieval step
Heat mediated
Permeabilization
No
Specification
Liver
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Formaldehyde

Muhammad Umair Latif

Verified customer

Submitted Jun 18 2019

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Hu pluripotent stem cell derived epicardial like)
Permeabilization
Yes - saponin
Specification
Hu pluripotent stem cell derived epicardial like
Blocking step
Serum as blocking agent for 15 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted May 06 2016

Answer

Anti-NFAT2 [7A6] antibody - ChIP Grade, ab2796 binds to the cytoplasmic form within amino acids 1-654.

Read More

Answer

Thank you for your enquiry.

I can confirm that ab2796 NFAT2 antibody [7A6] antibody is sold asascites fluid. Unpurified antibodies, such as those sold as whole antiserum, ascites or tissue culture supernatant will not have a concentration stated on the datasheet. Antibody concentration is usually determined by protein assay, and serum / ascites / tissue culture supernatant will contain a lot of other proteins, which means the antibody quantification would not be accurate.

I can confirm that for ascites, concentration of antibody is known to very between 5 - 10 mg/ml.

I am sorry we are not able to provide an exact concentration on this occasion, but hope this information will be helpful to you. If you have any further questions, please do not hesitate to contact us.

Read More

Answer

Thank you for your enquiry.

The lot we currently have available is #1312558 (GR73411-4).

I hope this information helps.

Read More
Application
ChIP
Sample
Human Cell lysate - nuclear (Primary Human T cells)
Specification
Primary Human T cells
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 4 minute(s) and 0 second(s)
Specification of the cross-linking agent: 37% Formaldehyde
Detection step
Real-time PCR
Positive control
Primary Human Regulatory T cells stimulated for 24 hours and analysed for NFAT recruitment to a predicted NFAT binding site
Negative control
Primary Human T effector cells stimulated for 24 hours and analysed for NFAT recruitment to the same NFAT binding site. Additionally, an AFM intronic region was also used as an additional negative control.

Abcam user community

Verified customer

Submitted Jun 20 2012

Application
Western blot
Sample
Mouse Cell lysate - whole cell (el4)
Loading amount
100000 cells
Specification
el4
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Jun 12 2012

Answer

Vielen Dank für diese Nachricht.

Wir freuen uns auf Ihre Rückmeldung - wannimmer Sie soweit sind :-)

Read More

Answer

Thank you for your inquiry. This antibody is unpurified ascites. The natural color of ascites fluid can vary extensively since it is taken directly from the peritoneal cavity. If "ascites" is stated in the purity section of the datasheet means the following: Monoclonal antibodies are produced by growing hybridoma cells within the peritoneal cavity of a mouse (or rat). When injected into a mouse, the hybridoma cells multiply and produce fluid (ascites) in its abdomen. This fluid contains a high concentration of antibody which can be harvested. This usually provides higher antibody yields than hybridoma cell culture. I happy to confirm that ab2796 (Lot GR24078-1) is good to use and there are no quality issues with this antibody. I hope this information is helpful. Please do not hesitate to contact me again with any further questions in this matter.

Read More

1-10 of 13 Abreviews or Q&A

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