Description
STATs (signal transducers and activators of transcription) are critical mediators of the biologic activity of cytokines including Interleukins (IL) 2-5, IL-7, IL-15, GM-CSF, erythropoietin and growth hormone.Ligand-receptor interaction leads to activation of constitutively associated JAK family kinases and subsequent recruitment/activation of STATs by tyrosine phosphorylation.Active STATs then move to the nucleus to promote transcription of cytokine-inducible genes.Seven STAT proteins have been cloned, each of which is differentially expressed and/or activated in a cytokine-specific and cell type-specific manner.Stat6 plays an important role in signaling pathways that lead to the differentiation of T helper type 2 (Th2) cells from uncommitted CD4 T cell precursors. Moreover, IL-4, secreted by activated T lymphocytes, basophils, and mast cells, induces specific gene expression via the induction of tyrosine phosphorylation of Stat6 at tyrosine 641 (Y641).The SH3:SH2 domain of Stat6 associates with tyrosine-phosphorylated IL-4 receptor and the proximal Jak kinase phosphorylates Stat6 at Y641 on the C-terminal side of the SH2 domain.Stat6 is then released from the receptor, dimerizes, and is thought to contact the basal transcription machinery by binding to p300/CBP.While Stat6 is widely expressed in human tissues, it exhibits elevated expression in peripheral blood lymphocytes, colon, intestine, ovary, prostate, thymus, spleen, kidney, liver, lung, and placenta.
The 23/Stat6 monoclonal antibody recognizes Stat6, regardless of phosphorylation status.
Format
- FormatPerCP-Cy™5.5
- Excitation SourceBlue 488 nm
- Excitation Max482nm
- Emission Max678nm
PerCP-Cy™5.5 is a tandem conjugate that combines PerCP with a cyanine dye. PerCP-Cy5.5 is not subject to photobeaching like PerCP and can be used with stream-in-air flow cytometers. It has less Fc receptor–mediated nonspecific staining than PE-Cy5. Additionally, the PerCP-Cy5.5 tandem conjugate is not as susceptible to fixative or light instability compared to APC-Cy7 and PE-Cy7.
Other Formats→
- Alexa Fluor® 647
- PE
- PE-CF594
- Purified
Suggested Companion Products
Stain Buffer (FBS) RUO 500mLCat No: 554656
PerCP-Cy™5.5 Mouse IgG1 κ Isotype Control RUO 0.1mgCat No: 550795
Fixation Buffer RUO 100mLCat No: 554655
Perm Buffer III RUO 125mLCat No: 558050
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Preparation and Storage
Store undiluted at 4°C and protected from prolonged exposure to light.Do not freeze.The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.The antibody was conjugated with PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed.Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test.We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Species testing during development may have been performed with a different format of the same clone.Selected applications have been tested for cross-reactivity.
- Please observe the following precautions:Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
- PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
- Cy is a trademark of Amersham Biosciences Limited.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
