WB result of AKT1 Rabbit mAb
Primary antibody: AKT1 Rabbit mAb at 1/5000 dilution
Lane 1: HeLa whole cell lysate 20 µg
Lane 2: HEK-293 whole cell lysate 20 µg
Lane 3: MCF7 whole cell lysate 20 µg
Lane 4: A549 whole cell lysate 20 µg
Lane 5: HepG2 whole cell lysate 20 µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 56 kDa
Observed MW: 60 kDa
Product Details
Product Details
Product Specification
Host | Rabbit |
Synonyms | RAC-alpha serine/threonine-protein kinase, Protein kinase B (PKB), Protein kinase B alpha (PKB alpha), Proto-oncogene c-Akt, RAC-PK-alpha, RAC, PKB |
Location | Cytoplasm, Nucleus, Cell membrane |
Accession | P31749 |
Clone Number | S-R364 |
Antibody Type | Recombinant mAb |
Isotype | IgG |
Application | WB, ICC, ICFCM |
Reactivity | Hu, Ms, Rt |
Purification | Protein A |
Concentration | 0.5 mg/ml |
Conjugation | Unconjugated |
Physical Appearance | Liquid |
Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
application | dilution | species |
WB | 1:5000 | null |
ICC | 1:500 | null |
ICFCM | 1:500 | null |
Background
RAC (Rho family)-alpha serine/threonine-protein kinase is an enzyme that in humans is encoded by the AKT1 gene. This enzyme belongs to the AKT subfamily of serine/threonine kinases that contain SH2 (Src homology 2-like) protein domains. In the developing nervous system AKT is a critical mediator of growth factor-induced neuronal survival. Survival factors can suppress apoptosis in a transcription-independent manner by activating the serine/threonine kinase AKT1, which then phosphorylates and inactivates components of the apoptotic machinery. AKT1 is catalytically inactive in serum-starved primary and immortalized fibroblasts. AKT1 and the related AKT2 are activated by platelet-derived growth factor. The activation is rapid and specific, and it is abrogated by mutations in the pleckstrin homology domain of AKT1.
Picture
Picture
Western Blot
FC
Flow cytometric analysis of 4% PFA fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling AKT1 antibody at 1/500 dilution (0.1 μg)/ (Red) compared with a Rabbit monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti - Rabbit IgG Alexa Fluor® 488 was used as the secondary antibody.
Immunocytochemistry
ICC shows positive staining in HeLa cells. Anti-AKT1 antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
