Detection of Human and Mouse Phospho-Akt1 (T308) by Western Blot.
Western blot shows lysates of Jurkat human acute T cell leukemia cell line and NIH‑3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with 100 ng/mL Recombinant Human PDGF‑BB (Catalog # 220-BB) for 20 minutes or 100nM Calyculin A for 30 minutes. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Phospho-Akt1 (T308) Monoclonal Antibody (Catalog # MAB7419) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Phospho-Akt1 (T308) at approximately 65 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Phospho-Akt1 (T308) in CCD‑1070Sk Human Cell Line.
Akt1 phosphorylated at T308 was detected in immersion fixed CCD‑1070Sk human foreskin fibroblast cell line stimulated with Recombinant Human PDGF‑BB (Catalog # 220-BB) using Mouse Anti-Human Phospho-Akt1 (T308) Monoclonal Antibody (Catalog # MAB7419) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; 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.
Akt, also known as protein kinase B (PKB), is a central kinase in such diverse cellular processes as glucose uptake, cell cycle progression, and apoptosis. Three highly homologous members define the Akt family: Akt1 (PKB alpha ), Akt2 (PKB beta ), and Akt3 (PKB gamma ). All three Akts contain an amino-terminal pleckstrin homology domain, a central kinase domain, and a carboxyl-terminal regulatory domain. Akt1 is the most widely expressed family member and is frequently activated in a number of carcinomas, including breast, prostate, lung, pancreatic, liver, ovarian, and colorectal cancer. Akt1 is activated in a multistep process that involves the sequential phosphorylation of Thr450 by JNK kinases, Thr308 by PDK1, and Ser473 by PDK2 or mTORC2. Activated Akt1 phosphorylates a wide variety of cytosolic, nuclear, and mitochondrial substrates. Human Akt1 shares 98% aa sequence identity with mouse and rat Akt1. MAB7419 also detects mouse Phospho-Akt1 (T308) in Western Blot.
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