抗AKT1 [p Ser473] (104A282)抗体 | Anti-AKT1 [p Ser473] (104A282) Antibody

• 価格
• Image
• Background

価格

目次に戻る

Image

	Independent Antibodies Validation and Biological Strategies Validation.Western Blot: AKT1 [p Ser473] Antibody (104A282) [NB100-56749] - Akt1 [p Ser473] Antibody (104A282) [NB100-56749] - Total protein from mouse 3T3 cells treated with and without PDGF (50 ng/ml) for the indicated times was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/ml anti-AKT1 (NBP2-01725) and 2 ug/ml pS473 AKT1 in 1% BSA in TBST and detected with an anti-mouse HRP secondary antibody using chemiluminescence. Note the detection of phosphorylated AKT1 in response to PDGF treatment compared to total AKT1 protein.
	Immunohistochemistry: AKT1 [p Ser473] Antibody (104A282) [NB100-56749] - Western Blot: AKT1 [p Ser473] Antibody (104A282) - Analysis of phospho AKT using phospho AKT antibody at 2 ug/ml against untreated (lane 1) and PDGF treated (lane 2) NIH-3T3 lysate. HRP conjugated secondary antibody and ECL substrate solution were used for this test. Image using the Azide and BSA Free form of this antibody.
	Immunohistochemistry-Paraffin: AKT1 [p Ser473] Antibody (104A282) [NB100-56749] - IHC analysis of a formalin-fixed paraffin-embedded (FFPE) human breast carcinoma tissue section using 1:250 dilution of pSer473 AKT1 antibody (clone 104A282) on a Bond Rx autostainer (Leica Biosystems). The assay involved 20 minutes of heat induced antigen retrieval (HIER) with 10mM sodium citrate buffer (pH 6.0) and endogenous peroxidase quenching using peroxide block. The sections were incubated with primary antibody for 30 minutes. Bond Polymer Refine Detection (Leica Biosystems) and DAB were used for signal detection which followed counterstaining with hematoxylin. Whole slide scanning and capturing of representative images (20X) were performed using Aperio AT2 (Leica Biosystems). This antibody generated a diffused cytoplasmic staining of phosphor-AKT (Ser-473) in the cancer cells as well as the stromal cells. Staining was performed by Histowiz.
	Immunohistochemistry-Paraffin: AKT1 [p Ser473] Antibody (104A282) [NB100-56749] - IHC analysis of a formalin-fixed paraffin-embedded (FFPE) human breast carcinoma tissue section using 1:250 dilution of pSer473 AKT1 antibody (clone 104A282) on a Bond Rx autostainer (Leica Biosystems). The assay involved 20 minutes of heat induced antigen retrieval (HIER) with 10mM sodium citrate buffer (pH 6.0) and endogenous peroxidase quenching using peroxide block. The sections were incubated with primary antibody for 30 minutes. Bond Polymer Refine Detection (Leica Biosystems) and DAB were used for signal detection which followed counterstaining with hematoxylin. Whole slide scanning and capturing of representative images (20X) were performed using Aperio AT2 (Leica Biosystems). This antibody generated a diffused cytoplasmic staining of phosphor-AKT (Ser-473) in the cancer cells as well as the stromal cells. Some cancer cells depicted nuclear stianing also. Staining was performed by Histowiz.

目次に戻る

Background

Akt, protein kinase B (PKB), is a serine/threonine kinase which is involved in many cellular signaling pathways and acts as a transducer of many functions initiated by growth factor receptors that activat phosphtidylinositol 3-kinase (PI 3-kinase). The major activity of Akt/PKB is to mediate cell survival. Akt/PKB is also believed to be a critical factor in the genesis of cancer as the tumor suppressor PTEN was found to antagonise PI-3 kinase and Akt/PKB kinase activity. Akt/PKB phosphorylation is critical for it activity. The major phosphorylation sites required for Akt activation has been identified as threoinine 308 and serine 473. Serine 473 is phosphorylated by MAPKAP kinase 2.

目次に戻る