SSTR3 Antikörper

Produktnummer ABIN7242716

Dieses unabhängig validierte Kaninchen Polyklonal-Antikörper erkennt spezifisch SSTR3 in IHC und ELISA. Er zeigt eine Reaktivität gegenüber Human.

Kurzübersicht für SSTR3 Antikörper (ABIN7242716)

Target: SSTR3 (Somatostatin Receptor 3 (SSTR3))

Reaktivität: Human

Wirt: Kaninchen

Klonalität: Polyklonal

Konjugat: Dieser SSTR3 Antikörper ist unkonjugiert

Applikation: Immunohistochemistry (IHC), ELISA

Produktdetails anti-SSTR3 Antikörper

Produktmerkmale: Polyclonal Antibody

Aufreinigung: Affinity purification

Immunogen: Synthetic peptide of human SSTR3

Isotyp: IgG

Anwendungsinformationen

Applikationshinweise: IHC 1:50-1:200

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (IF)

Validierung #103937 (Immunofluorescence)

by: Idevall Lab, Department of Medical Cell Biology, Uppsala University

No.: #103937

Datum: 18.06.2019

Antigen: SSTR3

Chargennummer: DK1445

Validierte Anwendung: Immunofluorescence

Positivkontrolle:

Pancreatic beta cell line MIN6, stained for acetylated tubulin

Bewertung:

ABIN7242714 specifically labels the targeted antigen in mouse insulinoma cells in IF. No signal was detected in sample negative control tissue and the secondary antibody only control.

Validierungsbilder

Staining of MIN6 cells with and antibody directed against acetylated tubulin (red, a bona fide marker of the cilium), ABIN7242714 (green) and DAPI (blue). Composite image shows the overlap of red and green staining in the cilium.

Fluorescence profiles measured along the cilium of the channels corresponding to acetylated tubulin (red) and SSTR3 (green) confirming enrichment of the receptor in the medial portion of the cilium.

Protokoll

Primärantikörper: ABIN7242714

Full Protocol:

• Grow MIN6 cells (gift from Prof. Yamamura) in DMEM (ThermoFisher, 41965039, lot 2071491) supplemented with FBS (ThermoFisher, 10500064, lot 08Q6291K) and Penicilin-Streptomycin (ThermoFisher, 15140122, lot 2051356), at 37 °C and 5% CO₂ in 2 mL on glass coverslips.
• Starve cells for 48 h in serum free medium to promote ciliogenesis.
• Wash cells with 37 °C DPBS (ThermoFisher, 14190094, lot 2062255).
• Fix cells on coverslips in 4% PFA for 2 min at RT.
• Wash cells 3x for 2 min with PBS.
• Permeabilize cells in PBS containing 0.1% Triton-X for 2 min at RT.
• Block non-specific binding with PBS containing 1% serum for 1 h at RT.
• Incubate cells with primary
• • rabbit anti-SSTR3 antibody (antibodies-online, ABIN7242714, lot DK1445) and
  • mouse anti-actetylated tubulin (Sigma-Aldrich, T7451, lot 078M4842V) diluted 1:100 in 1% BSA PBS for 2 h at RT.
• Wash cells 3x for 2 min with PBS.
• Incubate cells with secondary
• • goat anti-rabbit AF488-conjugated antibody (ThermoFisher, A11034, lot 2069632) and
  • goat anti-mouse AF568-conjugated antibody (ThermoFisher, A11004, lot 927620) diluted 1:200 in 1% BSA PBS for 1 h at RT.
• Wash cells 2x for 2 min with PBS.
• Mount coverslips on glass slides in ProLong Gold antifade reagent with DAPI (ThermoFisher, P36935, lot 2086914).
• Image acquisition with Zeiss LSM780, 60x magnification.

Anmerkungen:

Our results are consistent with previous studies that have revealed the exclusive expression of SSTR3 in the cilia of mouse pancreatic beta-cells (Iwanaga et al. (2011)).

Handhabung

Format: Liquid

Konzentration: 0.4 mg/mL

Buffer: PBS with 0.05 % sodium azide and 50 % glycerol, PH7.4

Konservierungsmittel: Sodium azide

Vorsichtsmaßnahmen: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Lagerung: -20 °C

Informationen zur Lagerung: Store at -20°C. Avoid freeze / thaw cycles.

Details zu SSTR3

Target: SSTR3 (Somatostatin Receptor 3 (SSTR3))

Andere Bezeichnung: SSTR3

Hintergrund: Somatostatin acts at many sites to inhibit the release of many hormones and other secretory proteins. The biological effects of somatostatin are probably mediated by a family of G protein-coupled receptors that are expressed in a tissue-specific manner. SSTR3 is a member of the superfamily of receptors having seven transmembrane segments and is expressed in highest levels in brain and pancreatic islets. SSTR3 is functionally coupled to adenylyl cyclase.

NCBI Accession: NP_001042

UniProt: P32745