Alexa Fluor® 647 Mouse anti-PDPK1 (pS241)
Clone J66-653.44.17 (RUO)
- Brand BD Phosflow™
- Alternative Name PDK1 (pS241), PKB Kinase (pS241)
- Vol. Per Test 20 µl
- Isotype Mouse IgG1, κ
- Reactivity Human (QC Testing) Mouse, Rat (Predicted)
Intracellular staining (flow cytometry) (Routinely Tested)
- Immunogen Phosphorylated Human PDPK1 Peptide
- Storage Buffer Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
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.
Alexa Fluor® 647 conjugates are highly photostable and remain fluorescent over a broad pH range. The excitation and emission maxima are nearly identical to those of APC. However, APC tends to be brighter while Alexa Fluor® 647 is more optimal for intracellular applications. This fluorochrome exhibits uncommon photostability, making it an ideal choice for use in fluorescence microscopy. Due to nearly identical excitation and emission properties but different spillover characteristics, APC and Alexa Fluor® 647 cannot be used simultaneously.
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Preparation and Storage
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
- 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).
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- 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.
- 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.
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