SSTR3 anticorps

N° du produit ABIN7242716

Cet validé indépendamment anticorps Lapin Polyclonal détecte spécifiquement SSTR3 dans IHC et ELISA. Il présente une réactivité envers Humain.

Aperçu rapide pour SSTR3 anticorps (ABIN7242716)

Antigène: SSTR3 (Somatostatin Receptor 3 (SSTR3))

Reactivité: Humain

Hôte: Lapin

Clonalité: Polyclonal

Conjugué: Cet anticorp SSTR3 est non-conjugé

Application: Immunohistochemistry (IHC), ELISA

Détail du produit anti-SSTR3 anticorps

Attributs du produit: Polyclonal Antibody

Purification: Affinity purification

Immunogène: Synthetic peptide of human SSTR3

Isotype: IgG

Information d'application

Indications d'application: IHC 1:50-1:200

Restrictions: For Research Use only

Independent Validation (IF)

Validation #103937 (Immunofluorescence)

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

No.: #103937

Date: 18.06.2019

Antigène: SSTR3

Numéro du lot: DK1445

Application validée: Immunofluorescence

Contrôle positif:

Pancreatic beta cell line MIN6, stained for acetylated tubulin

Conclusion:

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.

Validation Images

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.

Protocole

Anticorps primaire: 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.

Notes:

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)).

Stockage

Format: Liquid

Concentration: 0.4 mg/mL

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

Agent conservateur: Sodium azide

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

Stock: -20 °C

Stockage commentaire: Store at -20°C. Avoid freeze / thaw cycles.

Détails sur SSTR3

Antigène: SSTR3 (Somatostatin Receptor 3 (SSTR3))

Autre désignation: SSTR3

Sujet: 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