Detection of Human, Mouse, and Rat PDK‑1 by Western Blot.
Western blot shows lysates of Jurkat human acute T cell leukemia cell line, HepG2 human hepatocellular carcinoma cell line, Daudi human Burkitt's lymphoma cell line, A20 mouse B cell lymphoma cell line, and C6 rat glioma cell line. PVDF Membrane was probed with 1 µg/mL of Mouse Anti-Human/Mouse/Rat PDK‑1 Monoclonal Antibody (Catalog # MAB864) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). Specific bands were detected for PDK‑1 at approximately 58-68 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
PDK‑1 in HeLa Human Cell Line.
PDK‑1 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Mouse Anti-Human/Mouse/Rat PDK‑1 Monoclonal Antibody (Catalog # MAB864) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (yellow; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
Sterile PBS to a final concentration of 0.5 mg/mL.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
PDK-1 (3-phosphoinositide-dependent protein kinase, gene name PDPK1) is a 58 -68 kDa, 556 amino acid (aa) monomeric protein of the AGC serine/threonine kinase family. It is activated by phosphorylation in the presence of PtdIns(3,4,5) P3 or PtdIns(3,4) P2. Akt, S6 kinases, PKA and PKC-zeta are reported PDK-1 substrates. Through Akt, PDK-1 mediates many of the intracellular actions of insulin. Within the region used as an immunogen, human PDK-1 shares 98% aa identity with mouse and rat PDK-1. One reported isoform has an alternate start site at aa 50, while another lacking aa 238-263 is predicted to be catalytically inactive.
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