-
Your selected country is
Sweden
- Change country/language
Old Browser
This page has been recently translated and is available in French now.
Looks like you're visiting us from {countryName}.
Would you like to stay on the current country site or be switched to your country?
Analysis of PDPK1 (pS241) in human peripheral blood mononuclear cells (PBMC). PBMC were either treated with calyculin A plus okadaic acid for 30 minutes at 37ºC (solid line histograms) or untreated (dashed line histograms). The cells were fixed with pre-warmed BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049) for 15 minutes at 37ºC and permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050) on ice for 30 minutes. The treated and untreated PBMC were stained with PE Mouse IgG1, κ isotype control mAb MOPC-21 (Cat. No. 559320, left panel) or PE Mouse anti-PDPK1 (pS241, Cat. No. 560092, right panel). The data demonstrates that the phosphorylation of PDPK1 is constitutive in all PBMC and that the level of phosphorylation increases when phosphatase activity is inhibited by the treatment. Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.
BD™ Phosflow PE Mouse anti-PDPK1 (pS241)
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
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 Jurkat Calyculin A + Okadaic Acid Lyse/Fix or Cytofix III Up-regulation
Flow Human Jurkat Calyculin A + Okadaic Acid Lyse/Fix I or II Unsatisfactory
Flow Human PBMC Calyculin A + Okadaic Acid Lyse/Fix III Up-regulation
WB Human Jurkat Calyculin A + Okadaic Acid 63 kDa
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.
- 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.
- 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/us/s/resources for technical protocols.
Companion Products
The serine/threonine kinase 3-Phosphoinositide-Dependent Protein Kinase-1 (PDPK1, also known as PDK1) contributes to the activation of many important kinases in the insulin and IGF-1 signaling pathways. It acts downstream of phosphatidylinositol 3-kinase (PI3-kinase) to phosphorylate residues in the activation loops of many cellular kinases, including protein kinase B (PKB/Akt), PKC isoforms, p70 S6 kinase, and PDPK1 itself. The autophosphorylation of PDPK1 at serine 241 (S241) has recently been suggested to play a role in the regulation of PDPK1. It has been proposed that PDPK1 activity plays a key role in the regulation of various cellular events such as cell proliferation, differentiation, and apoptosis.
The J666-653.44.17 monoclonal antibody recognizes the phosphorylated S241 in the activation loop of human PDPK1. The orthologous phosphorylation site in mouse and rat PDPK1 is S244.
Development References (2)
-
Komander D, Kular G, Deak M, Alessi DR, van Aalten DM. Role of T-loop phosphorylation in PDK1 activation, stability, and substrate binding. J Biol Chem. 2005; 280(19):18797-18802. (Biology). View Reference
-
Wick MJ, Ramos FJ, Chen H, et al. Mouse 3-phosphoinositide-dependent protein kinase-1 undergoes dimerization and trans-phosphorylation in the activation loop. J Biol Chem. 2003; 278(44):42913-42919. (Biology). View Reference
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.