Recombinant
RabMAb

Recombinant Anti-STAT6 (phospho Y641) antibody [EPR22599-52] - BSA and Azide free (ab263949)

Overview

  • Product name

    Anti-STAT6 (phospho Y641) antibody [EPR22599-52] - BSA and Azide free
    See all STAT6 primary antibodies
  • Description

    Rabbit monoclonal [EPR22599-52] to STAT6 (phospho Y641) - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ChIP, Flow Cyt, IP, Dot blotmore details
    Unsuitable for: ICC/IF or IHC-P
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Mouse STAT6 aa 600-700. The exact sequence is proprietary.
    Database link: P52633

  • Positive control

    • WB and IP: Daudi serum starved for 24 hours, then treated with 100 ng/ml IL-4 for 15 minutes, whole cell lysate; 2.4G2 serum starved for 24 hours, then treated with 100 ng/ml rIL-4 for 15 minutes, whole cell lysate; RAW264.7 serum starved for 24 hours, then treated with 100 ng/ml mIL-4 for 15 minutes, whole cell lysate. Flow: Daudi was serum starved for 24h, then treated with 100ng/ml IL-4 for 15min; RAW 264.7 was serum starved for 24h, then treated with 100ng/ml IL-4 for 15min
  • General notes

    ab263949 is the carrier-free version of ab235591. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.  

    ab263949 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab263949 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.

 

 

Flow Cyt Use at an assay dependent concentration.

 

 

IP Use at an assay dependent concentration.

 

 

Dot blot Use at an assay dependent concentration.

 

 

  • Application notes
    Is unsuitable for ICC/IF or IHC-P.
  • Target

    • Function

      Carries out a dual function: signal transduction and activation of transcription. Involved in interleukin-4 signalling.
    • Sequence similarities

      Belongs to the transcription factor STAT family.
      Contains 1 SH2 domain.
    • Post-translational
      modifications

      Tyrosine phosphorylated following stimulation by IL-4 and IL-3.
    • Cellular localization

      Cytoplasm. Nucleus. Translocated into the nucleus in response to phosphorylation.
    • Information by UniProt
    • Database links

    • Alternative names

      • 12S1644 antibody
      • D12S1644 antibody
      • IL 4 STAT antibody
      • IL-4 Stat antibody
      • IL4 STAT antibody
      • Interleukin 4 Induced antibody
      • Interleukin 4 Induced Transcription Factor IL4 STAT antibody
      • Signal transducer and activator of transcription 6 antibody
      • Signal Transducer And Activator Of Transcription 6 Interleukin 4 Induced antibody
      • Signal Transducer And Activator Of Transcription 6 Nirs Variant 1 antibody
      • Signal transducer and activator of transcription 6, interleukin 4 induced antibody
      • STAT 6 antibody
      • STAT interleukin4 induced antibody
      • STAT, interleukin4 induced antibody
      • Stat6 antibody
      • STAT6_HUMAN antibody
      • STAT6B antibody
      • STAT6C antibody
      • Transcription factor IL 4 STAT antibody
      see all

    Images

    • Chromatin was prepared from Daudi cells starved for 24h, then treated with hIL-4(100ng/ml, 15min) according to the Abcam Dual-X-ChIP protocol*. Cells were fixed with 1.5 mM EGS for 30mins and then formaldehyde for 10min. 
      The ChIP was performed with 25 µg of chromatin, 5 µg of ab235591 (red), or 5 µg of rabbit normal IgG ab172730 (gray) and 20 µl of Protein A/G sepharose beads. The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci). 
      Primers and probes are commercial primers from Cell Signaling Technology (Cat. No.: #56554) 
      *https://www.abcam.com/resources?keywords=X%20ChIP%20protocol

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab235591).

    • Dot blot analysis using ab235591 at 1/1000 dilution, Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) was used as the secondary antibody at 1/100,000 diluton.

      Lane 1: STAT6 (phospho Y641) peptide (aa636-647).

      Lane 2: STAT6 non-phospho peptide (aa636-647)

      Exposure time: 26 seconds.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab235591).

    • STAT6 (phospho Y641) was immunoprecipitated from 0.35 mg RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) cells serum starved for 24h, then treated with 100ng/ml IL-4 for 15min whole cell lysate with ab235591 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab235591 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

      Lane 1: RAW 264.7 cells serum starved for 24h, then treated with 100ng/ml IL-4 for 15min whole cell lysate 10ug

      Lane 2: ab235591 IP in RAW 264.7 cells serum starved for 24h, then treated with 100ng/ml IL-4 for 15min whole cell lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab235591 in RAW 264.7 was serum starved for 24h, then treated with 100ng/ml IL-4 for 15min whole cell lysate

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 10 seconds

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab235591).

    • STAT6 (phospho Y641) was immunoprecipitated from 0.35 mg Daudi (Human Burkitt's lymphoma lymphoblast) cells serum starved for 24h, then treated with 100ng/ml IL-4 for 15min whole cell lysate, with ab235591 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab235591 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

      Lane 1: Daudi cells serum starved for 24h, then treated with 100ng/ml IL-4 for 15min whole cell lysate 10ug

      Lane 2: ab235591 IP in Daudi was serum starved for 24h, then treated with 100ng/ml IL-4 for 15min whole cell lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab235591 in Daudi cells serum starved for 24h, then treated with 100ng/ml IL-4 for 15min whole cell lysate

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 10 seconds.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab235591).

    • Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) cells was serum starved for 24h, then treated with 100ng/ml IL-4 for 15min (Red) / Untreated control (Green) cells labelling STAT6 (phospho Y641) with ab235591 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1:2000 dilution was used as the secondary antibody.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab235591).

    • Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Daudi (Human Burkitt's lymphoma lymphoblast) was serum starved for 24h, then treated with 100ng/ml IL-4 for 15min (Red) / Untreated control (Green) cells labelling STAT6 (phospho Y641) with ab235591 at 1/500 (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1:2000 dilution was used as the secondary antibody.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab235591).

    References

    ab263949 has not yet been referenced specifically in any publications.

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