PE Mouse Anti-Stat5 (pY694)
Clone 47/Stat5 (pY694) (RUO)
- Brand BD Phosflow™
- Alternative Name Signal transducer and activator of transcription 5; MGF; MPF
- Vol. Per Test 20 µl
- Isotype Mouse IgG1, κ
- Reactivity Human (QC Testing) Rat, Mouse, Sheep (Predicted)
- Application
Intracellular staining (flow cytometry) (Routinely Tested)
- Immunogen Phosphorylated Human Phosphorylated Stat5 Peptide
- Storage Buffer Aqueous buffered solution containing BSA, protein stabilizer, and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
Description
Stat (Signal transducer and activators of transcription) proteins mediate the biological activity of cytokines, including interleukins, interferons, erythropoietin, and growth factors. Ligand-receptor interaction activates constitutively-associated JAK family kinases and subsequent recruitment/activation of Stat proteins by tyrosine phosphorylation. Active Stat proteins 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. Stat5 has been characterized and shown to be encoded by two separate genes, Stat5a and Stat5b, which share over 90% identity at the amino acid level. Stat5a has been shown to be involved in lactogenesis and mammary development, while Stat5b has been shown to be involved in growth hormone signaling and liver gene expression. Both Stat5a and Stat5b are involved in IL-2 induced peripheral T cell proliferation. The peptide hormone, prolactin, binds to the prolactin receptor (PRLR) to initiate the lactogenic response. There are at least three forms of PRLR; however, only the long form activates the 92-kDa Stat5 protein by inducing phosphorylation at Y694. Once phosphorylated, Stat5 becomes an essential transcription factor which binds to the β-casein gene promoter. The presence of an SH2 domain within Stat5 suggests that it may directly interact with protein tyrosine kinases (PTKs) such as JAK2.
The 47 monoclonal antibody recognizes the phosphorylated Y694 of Stat5a. The homologous phosphorylation site in Stat5b is Y699.
Format
- Format PE
- Excitation Source Blue 488 nm,Green 532 nm,Yellow/Green 561 nm
- Excitation Max 496 nm
- Emission Max 578 nm
R-phycoerythrin (PE) is an accessory photosynthetic pigment found in red algae. It exists in vitro as a 240-kDa protein with 23 phycoerythrobilin chromophores per molecule. This makes PE one of the brightest fluorochromes for flow cytometry applications, but its photobleaching properties make it unsuitable for fluorescence microscopy.
Suggested Companion Products
Perm Buffer III RUO
125 mL
Cat No: 558050
Fix Buffer I RUO
250 mL
Cat No: 557870
Fixation Buffer RUO
100 mL
Cat No: 554655
Purified Mouse Anti-Human Stat5 (pY694) 47/Stat5 (pY694) RUO
50 µg
Cat No: 611964
Lyse/Fix Buffer 5X RUO
250 mL
Cat No: 558049
Recombinant Human IL-2 RUO
10 µg
Cat No: 554603
Stain Buffer (FBS) RUO
500 mL
Cat No: 554656
Stain Buffer (BSA) RUO
500 mL
Cat No: 554657
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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 R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
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).
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- 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.
This antibody conjugate is suitable for intracellular staining of human whole blood (using BD Phosflow Lyse/Fix Buffer) and peripheral blood mononuclear cells (using BD Cytofix™ Fixation Buffer or BD Phosflow Fix Buffer I).
This mAb was characterized by flow cytometry (Flow) and western blot analysis (WB) using these model systems:
Method Species Cells Treatment Fixation Perm buffer Result
Flow Human PBMC IL-2 Fixation Buffer III Positive Staining
Flow Human PBMC IL-2 Fixation Buffer I or II Unsatisfactory
Flow Human Whole Blood IL-2 Lyse/Fix III Positive Staining
Flow Human Whole Blood IL-2 Lyse/Fix I or II Unsatisfactory
Flow Human TF-1 cells GM-CSF Fixation Buffer III Positive Staining
Flow Human TF-1 cells GM-CSF Fixation Buffer I or II Unsatisfactory
WB Human A431 Cell Lysate EGF Not Applicable Not Applicable 92 kDa