AKT Phospho specific pS474 Antikörper

Antigen: AKT Phospho specific pS474

Klonalität: Polyklonal

Wirt: Kaninchen

Reaktivität: Human, Rind (Kuh), Maus, Ratte (Rattus)

Applikation: Immunhistochemie (IHC), ELISA, Western Blot (WB), Immunpräzipitation (IP)

Produktnummer: ABIN94784

Produktbeschreibung

Immunogen: This whole rabbit serum was prepared by repeated immunizations with a synthetic peptide corresponding to the C-terminus aa 460-480 of human, mouse, rat and chicken AKT proteins conjugated to KLH. Peptide Sequence: C-H-F-P-Q-F-pS-Y-S-A-S

Format: Liquid (sterile filtered)

Beschreibung: AKT is a component of the PI-3 kinase pathway and is activated by phosphorylation at Ser 473 and Thr 308. AKT is a cytoplasmic protein also known as Protein Kinase B (PKB) and rac (related to A and C kinases). AKT is a key regulator of many signal transduction pathways. AKT Exhibits tight control over cell proliferation and cell viability. Overexpression or inappropriate activation of AKT is noted in many types of cancer. AKT mediates many of the downstream events of PI 3-kinase (a lipid kinase activated by growth factors, cytokines and insulin). PI 3-kinase recruits AKT to the membrane, where it is activated by PDK1 phosphorylation. Once phosphorylated, AKT dissociates from the membrane and phosphorylates targets in the cytoplasm and the cell nucleus. AKT has two main roles: (i) inhibition of apoptosis, (ii) promotion of proliferation.

Spezifität: This product was prepared from monospecific antiserum by immunoaffinity chromatography using phospho peptide coupled to agarose beads followed by solid phase adsorption(s) against non-phospho peptide and non-specific peptide to remove any unwanted reactivities. Assay by immunoelectro-phoresis resulted in a single precipitin arc against anti-Rabbit Serum. This antibody is specific for phosphorylated human AKT. Minimal reactivity occurs against non-phosphorylated AKT. Reactivity against AKT from other species may occur but has not yet been tested.

Anwendungen

Applikationshinweise: Recommended Dilutions: This product was assayed against the peptide immunogen in a standard sandwich ELISA using Peroxidase conjugated anti-Rabbit IgG [H&L] (Goat) code ABIN101990 and ABTS (2,2-azino-bis-[3-ethylbenthiazoline-6-sulfonic acid]) code ABTS-100 as a substrate for 30 minutes at room temperature. A working dilution of 1:20,000 to 1:70,000 is suggested for this product. A single band of the expected apparent molecular weight was observed by immunoblot using a 1:500 dilution of the antibody. For immunohistochemistry use at a dilution of 1:100. No pre-treatment of sample is required. Optimal titers for other applications should be determined by the researcher. Note: This antibody is phospho specific for pT308 and is suitable for western blotting, immuno- histochemistry (formalin-fixed paraffin-embedded sections), immunofluorescence and ELISA.

Konzentration: 1.7 mg/ml (by UV absorbance at 280 nm)

Buffer: Stabilizer: None. Preservative: 0.01% (w/v) Sodium Azide. Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2.

Lagerung: Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use. Expiration date is one (1) year from date of opening.

Forschungsgebiet: Kinasen/Phosphatasen

Beschränkungen: Nur für Forschungszwecke einsetzbar

Bilder

Immunohistochemistry. Rabbit anti-AKT pS473 was used at a 1:100 dilution to detect AKT by immunohistochemistry in human breast tumor tissue. The staining is much stronger than the weak basal level of phosphorylation in normal breast. antibodies-online's Rabbit anti-AKT pS473 antibody was used with no pretreatment of tissue. Signal was developed using Dako's Techmate streptavidinbiotin reagents. Tissue was formalin-fixed and paraffin embedded.

Immunohistochemistry. This image is a higher magnification of the breast tumor image above.

anti-AKT Phospho specific pS474 antibody (Image 3)