Detection of Human Akt by Western Blot.
Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, Jurkat human acute T cell leukemia cell line, HepG2 human hepatocellular carcinoma cell line, MCF‑7 human breast cancer cell line, and KG‑1 human myeloid leukemia cell lines. PVDF membrane was probed with 0.1 µg/mL of Human Akt Pan Specific, Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2055) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Akt Pan Specific, at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
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.
Rabbit antibodies were raised against a synthetic peptide corresponding to amino acids 75‑90 of human v-akt murine thymoma viral oncogene homolog 1 (Akt1), also known as protein kinase B alpha (PKB alpha ) and RAC-alpha serine/threonine kinase (RAC-PK-alpha ). In addition, this peptide sequence is identical to amino acids 75‑90 of human Akt2 (PKB beta, RAC-PK-beta ), and amino acids 74‑89 of human Akt3 (PKB gamma, RAC-PK-gamma ).
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We have 1 review tested in 1 species: Human.
We have 1 review tested in 1 application: Western Blot.
Western Blot: Human Akt Pan Specific Antibody [AF2055].
Human Chronic Lymphocytic Cell Line
Other Experimental Details
Other Experimental Details
30 microg cell homogenate/lineGel SDS/Page 4-12% BisTris Novex Invitrogen.Western Blot on PVDF membrane.Blocking Reagent: 5% non fat dry milk/T-TBSAnti AKT1-2-3 (R& D System cat. N. AF2055) 1:1250 in 3% BSA/PBS o.n. 4°CAnti pAKTser473 Cell Signalling 1:1250 in 3%BSA/PBS o.n. 4°C.Anti horsadish peroxidase linked II antibody against Rabbit 1:20.000 (GE Healthcare) in T-TBS 90 min r.t.ECL Prime Amersham (GE Healhtcare).