Anti-NFAT2 antibody [7A6]

• 产品名称

  Anti-NFAT2抗体[7A6]
  参阅全部 NFAT2 一抗

• 描述

  小鼠单克隆抗体[7A6] to NFAT2

• 宿主

  Mouse

• 经测试应用

  适用于: WB, IHC-P, Flow Cyt (Intra)more details

• 种属反应性

  与反应: Human
  预测可用于: Mouse, Rat, Hamster, Non human primates

• 免疫原

  Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

• 阳性对照

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

• 常规说明

  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.

  
  

  The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

  If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

• 形式

  Liquid

• 存放说明

  Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.

• 存储溶液

  Preservative: 0.05% Sodium azide
  Constituent: PBS

• 浓度

• 10 µg 浓度为 0.5 mg/ml

• 100 µg 浓度为 0.5 mg/ml

• 纯度

  Affinity purified

• 克隆

  单克隆

• 克隆编号

  7A6

• 同种型

  IgG1

• 轻链类型

  kappa

The Abpromise guarantee

Abpromise™承诺保证使用ab2796于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

数据库链接

• Entrez Gene: 4772 Human

• Entrez Gene: 18018 Mouse

• Entrez Gene: 100361818 Rat

• Omim: 600489 Human

• SwissProt: O95644 Human

• SwissProt: O88942 Mouse

• Unigene: 534074 Human

• Unigene: 701518 Human

• Unigene: 329560 Mouse

别名

• 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

• 

  Western blot - Anti-NFAT2 antibody [7A6] (ab2796)

  All lanes : Anti-NFAT2 antibody [7A6] (ab2796) at 5 µg/ml
  
  Lane 1 : HeLa cell lysate
  Lane 2 : Wild-type HAP1 cell lysate
  Lane 3 : NFATC1 knockout HAP1 cell lysate
  
  Lysates/proteins at 20 µg per lane.
  
  Performed under reducing conditions.
  
  Observed band size: 75,80,90 kDawhy is the actual band size different from the predicted?
  
  
  
  

  False colour image of Western blot: Anti-NFAT2 antibody [7A6] staining at 5 ug/ml, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab2796 was shown to bind specifically to NFAT2. A band was observed at 75/80/90 kDa in wild-type HAP1 cell lysates with no signal observed at this size in NFATC1 knockout cell line. To generate this image, wild-type and NFATC1 knockout HAP1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFAT2 antibody [7A6] (ab2796)

  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

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFAT2 antibody [7A6] (ab2796)

  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.

• 

  Flow Cytometry (Intracellular) - Anti-NFAT2 antibody [7A6] (ab2796)

  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.