Alexa Fluor® 488 Mouse anti-PKCα (pT497)
Clone K14-984 (RUO)
- Brand BD Phosflow™
- Vol. Per Test 20 µl
- Isotype Mouse BALB/c IgG1, κ
- Reactivity Human (QC Testing) Mouse (Tested in Development)
Intracellular staining (flow cytometry) (Routinely Tested)
- Immunogen Phosphorylated Human PKCα Peptide
- Storage Buffer Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
The Protein Kinase C (PKC) family of serine/threonine protein kinases is involved in a number of processes such as growth, differentiation, and cytokine secretion. Three categories exist, conventional PKC (cPKC), novel PKC (nPKC), and atypical PKC (aPKC). All have C-terminal kinase domains, which are closely related to those of protein kinases A and B (Akt), and variable N-terminal regulatory domains that give them different modes of activation. For example, cPKC (α, βI, βII, and γ isoforms) are calcium-activated, phospholipid-dependent serine/threonine-specific enzymes that can also be activated by phorbol esters. However, nPKC (δ, ε, η, and θ isoforms) and aPKC (ζ, ι, and λ isoforms) are Ca2+-independent. aPKC are unique in that their activity is independent of diacylglycerols and phorbol esters. Phosphorylation at three conserved sites in the kinase domain is required for catalytic activity. Specifically, the threonine 497 (T497) of PKCα is in the activation loop of the kinase domain and is phosphorylated by the constitutively active phosphoinositide-dependent kinase-1 (PDK-1).
The K14-984 monoclonal antibody recognizes the phosphorylated T497 in the kinase domain of human PKCα.
Alexa Fluor® conjugates are highly photostable and remain fluorescent over a broad pH range. The excitation and emission maxima are nearly identical to those of FITC. However, Alexa Fluor® 488 tends to be brighter and more optimal for intracellular applications. Due to nearly identical excitation and emission properties but different spillover characteristics, FITC and Alexa Fluor® 488 cannot be used simultaneously. Alexa Fluor® 488 exhibits extraordinary photostability, which makes it highly suitable for fluorescence microscopy.
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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 to Alexa Fluor® 488 under optimum conditions, and unreacted Alexa Fluor® 488 was removed.
- 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.
- 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.
- Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- 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.