V-Akt Murine Thymoma Viral Oncogene Homolog 1 (AKT1) (pS473) Antikörper

Antigen: V-Akt Murine Thymoma Viral Oncogene Homolog 1 (AKT1)

Synonyme: AKT, PKB, RAC, PRKBA, MGC99656, PKB-ALPHA, RAC-ALPHA, Akt, pkb, xAct, akt-1, v-akt, v-akt1, AKT1, AKT/PKB, D-Akt, DAKT1, DAKT1/PKB, DPKB, DRAC-PK, DRAC-PK66; DRAC-PK85, Dakt, PKB/AKT, PKB/dAKT, RacPK, dAKT/dPKB, dAkt/PKB, l(3)04226, l(3)89Bq, DmelCG4006, CG4006

Epitope: pS473

Klonalität: Monoklonal (14-Mai)

Wirt: Kaninchen

Reaktivität: Human

Applikation: Immunhistochemie (IHC)

Zertifikate: ISO 9001:2008, ISO 13485:2003

Anbieter: Dako

Produktnummer: ABIN370673

Produktbeschreibung

Weitere Bezeichnung: Akt pS473

Immunogen: Synthetic phosphopeptide derived from the region of human Akt containing serine 473 Precautions

Klon: 14-Mai

Beschreibung: Akt, also known as protein kinase B (PKB) or Rac-alpha, is a serine/threonine protein kinase that functions as an important regulator of various cell processes including apoptosis, proliferation, differentiation and metabolism. Akt is a critical downstream effector of PI3-kinase (PI-3K), which mediates signal transduction initiated by a variety of stimuli including hormones, growth factors and cytokines. PI-3K activates Akt through a second messenger, which results in phosphorylation of Akt at threonine 308 and at serine 473 by upstream protein kinases. Activated Akt phosphorylates a number of protein substrates including BAD, caspase-9, Forkhead Transcription Factors, GSK-3-alpha –beta, CREB and mTOR/FRAP. 1, 2 Specificity

Spezifität: Anti-Akt-pS473, clone 14-5 recognizes a protein of approximately 60 kD in Western blots of Platelet Derived Growth Factor (PDGF) stimulated and unstimulated NIH3T3 cell extracts. The single band identified by clone 14-5 was very weakly stained on the immunoblot of the unstimulated NIH3T3 cell extract, whereas intense staining of the same band was observed on the blot of the PDGF stimulated cell extract. Calf intestinal alkaline phosphatase (CIP) treatment of formalin-fixed and paraffin-embedded (FFPE) tissue and AU-565 cell block sections (both known to be positive for Akt-pS473) resulted in a lack of immunostaining, demonstrating that the antibody reacts with a phosphorylated epitope. In peptide blocking studies, specific immunohistochemical staining with anti-Akt-pS473 was completely blocked when using antibody that was liquid phase absorbed with Akt-pS473 peptide on FFPE breast carcinoma tissue and AU-565 cell block sections known to be positive for Akt-pS473. No blocking effect was observed when anti-Akt-pS473 was combined with an unphosphorylated Akt peptide, demonstrating specificity of the antibody for the phosphorylated protein. provided

Synonyme: AKT, PKB, RAC, PRKBA, MGC99656, PKB-ALPHA, RAC-ALPHA, Akt, pkb, xAct, akt-1, v-akt, v-akt1, AKT1, AKT/PKB, D-Akt, DAKT1, DAKT1/PKB, DPKB, DRAC-PK, DRAC-PK66; DRAC-PK85, Dakt, PKB/AKT, PKB/dAKT, RacPK, dAKT/dPKB, dAkt/PKB, l(3)04226, l(3)89Bq, DmelCG4006, CG4006

Anwendungen

Applikationshinweise: This antibody is intended for laboratory use to identify qualitatively by light microscopy PTEN expressing cells in normal and neoplastic tissues using immunohistochemical (IHC) test methods. Differential identification is aided by the results from a panel of antibodies. Interpretation must be made within the context of the patient's clinical history and other diagnostic tests by a qualified pathologist. Paraffin Sections Anti-PTEN can be used on formalin-fixed, paraffin-embedded tissue sections. Pretreatment of tissue with proteolytic enzymes is not recommended. The deparaffinized tissue sections must be treated with heat prior to the IHC staining procedure. Heat-induced epitope retrieval (HIER) involves immersion of tissue sections in a pre-heated buffer solution and maintaining heat in a water bath (95-99°C). Alternative heat sources may be used for HIER upon validation against the recommended procedure. Use a 40-minute heating protocol for HIER performed at 95-99°C, after thermal treatment, allow the jar with buffer and slides to cool for 20 minutes at room temperature. Rinse well with buffer or deionized water following HIER. For greater adherence of tissue sections to glass slides, the use of Silanized Slides (code S3003) is recommended. Target Retrieval Solution pH 9.0 (code S2368) or 10x Concentrate (code S2367) is recommended. Cryostat Sections And Cell Smears Anti-PTEN can be used for labeling acetone-fixed cryostat sections or fixed cell smears. Follow the procedure for the detection system selected. Staining interpretation The cellular staining pattern for anti-PTEN is cytoplasmic and/or nuclear. (113878-001) 306645EFG_110905 p. 3/11 Normal Tissues 13 Tissue Type (# tested) Positively Staining Tissue Elements Adrenal (3) 2/3 zona glomerulosa, 2/3 vascular endothelium Bone Marrow (3) 1/3 macrophages Breast (3) 2/3 ductal epithelial cells, 1/3 lobular epithelial cells, 1/3 vascular endothelium Brain/Cerebellum (3) 3/3 molecular and granular layer, neuropil Brain/Cerebrum (3) 3/3 neurons, astrocytes, 2/3 neuropil Cervix (3) 3/3 squamous mucosa, 1/3 vascular smooth muscle, vascular endothelium Colon (2) 1/2 glandular epithelium, ganglion cells, macrophages, 1/2 germinal center cells, 2/2 nerve, vascular endothelium Esophagus (3) 1/3 squamous mucosa, 2/3 nerve, 1/3 vascular endothelium Heart (3) 3/3 vascular endothelium,1/3 vascular smooth muscle, 1/3 nerve Kidney (3) 2/3 glomeruli, tubule epithelium, endothelial cells, 1/3 mesangial cells, vessels, 1/3 lining cells, interstitial cells Liver (3) 3/3 hepatocytes, 1/3 bile ducts Lung (3) 2/3 bronchial epithelium, 3/3 vascular endothelium, 1/3 vascular smooth muscle, macrophages Mesothelial Cells (3) 3/3 vascular endothelial cells Nerve, Peripheral (2) 2/2 nerve cells Ovary (3) 3/3 stroma, 1/3 surface epithelium Pancreas (3) 2/3 ductal epithelium, nerve, 1/3 acinar epithelium, 3/3 islets of Langerhans,, 2/3 vascular endothelium Parathyroid (3) 2/3 chief cells Pituitary (3) 3/3 pituicytes, neuropil, 2/3 vascular endothelium, 1/3 nerve, vascular smooth muscle Prostate (3) 3/3 glandular epithelium, fibromuscular stroma, 1/3 vascular endothelium Salivary gland (3) 1/3 acini and duct epithelium, 2/3 nerve Skeletal muscle (3) 3/3 vascular endothelium, 1/3 vascular smooth muscle Skin (3) 3/3 epidermis, hair follicles, sebaceous glands, 1/3 apocrine glands Small intestine (3) 1/3 glandular epithelium, smooth muscle, macrophages, germinal center cells, 1/3 surface epithelium, 3/3 nerve, 3/3 vascular endothelium Spleen (3) 1/3 macrophages (rare), 1/3 littoral cells Stomach (3) 3/3 glandular epithelium, 2/3 vascular endothelium, 1/3 vascular smooth muscle, macrophages, 1/3 nerve, 1/3 germinal center cells Testis (3) 3/3 seminiferous tubules, 1/3 Leydig cells, vascular endothelium Thymus (3) 3/3 thymic epithelium Thyroid (2) 2/2 negative Tonsil (3) 3/3 squamous epithelium, 2/3 germinal center cells Uterus (3) 3/3 endometrial glands, endometrial stroma, myometrium Abnormal Tissues 11, 12, 14-16 Tumor Type (# tested) Positively Staining Tumors Breast carcinoma, sporadic primary carcinoma (33) 28/33 Colorectal carcinoma (90) 82/90 MSI + HNPCC (45) 40/45 MSI+ sporadic colorectal carcinomas (22) 19/22 MSI- sporadic colorectal carcinomas (23) 23/23 Endometrial cancer (72) 50/72 Pancreatic endocrine tumors, sporadic (24) 23/24 Thyroid carcinoma, sporadic nonmedullary (91) 83/91 Follicular thyroid carcinoma (28) 27/28 Papillary thyroid carcinoma (37) 35/37 Undifferentiated thyroid carcinoma (26) 21/26 (113878-001) 306645EFG_110905 p. 4/11

Buffer: Monoclonal Rabbit antibody provided in liquid form as cell culture supernatant dialysed against 0.05 mol/L Tris/HCl, pH 7.2, and containing 0.015 mol/L sodium azide. This product contains stabilizing protein.

Lagerung: Store at 2-8 °C. Precautions: 1. The device is not intended for clinical use including diagnosis, prognosis, and monitoring of a disease state, and it must not be used in conjunction with patient records or treatment. 2. For professional users. 3. This product contains sodium azide (NaN 3 ), a chemical highly toxic in pure form. At product concentrations, though not classified as hazardous, sodium azide may react with lead and copper plumbing to form highly explosive build-ups of metal azides. Upon disposal, flush with large volumes of water to prevent metal azide build-up in plumbing. 4. As with any product derived from biological sources, proper handling procedures should be used. 5. Wear appropriate Personal Protective Equipment to avoid contact with eyes and skin. 6. Unused solution should be disposed of according to local, State and Federal regulations.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Publikationen

Publikationen: Kadota, Yang, Gomez et al.: "Delineating genetic alterations for tumor progression in the MCF10A series of breast cancer cell lines." in: PLoS ONE, Vol. 5, Issue 2, pp. e9201, 2010 (PubMed).