SSTR3 antibody

Catalog No. ABIN7242716

Product Details anti-SSTR3 Antibody

Target: SSTR3 (Somatostatin Receptor 3 (SSTR3))

Reactivity: Human

Host: Rabbit

Clonality: Polyclonal

Conjugate: This SSTR3 antibody is un-conjugated

Application: Immunohistochemistry (IHC), ELISA

Characteristics: Polyclonal Antibody

Purification: Affinity purification

Immunogen: Synthetic peptide of human SSTR3

Isotype: IgG

Application Details

Application Notes: 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: 06/18/2019

Antigen: SSTR3

Lot Number: DK1445

Method validated: Immunofluorescence

Positive Control:

Pancreatic beta cell line MIN6, stained for acetylated tubulin

Notes:

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.

Full Methods

Primary Antibody: 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.

Experimental 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)).

Handling

Format: Liquid

Concentration: 0.4 mg/mL

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

Preservative: Sodium azide

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

Storage: -20 °C

Storage Comment: Store at -20°C. Avoid freeze / thaw cycles.

Target Details for SSTR3

Target: SSTR3 (Somatostatin Receptor 3 (SSTR3))

Alternative Name: SSTR3 (SSTR3 Products)

Synonyms: SS3R antibody, SS-3-R antibody, SS3-R antibody, SSR-28 antibody, Smstr-3 antibody, Smstr3 antibody, sst3 antibody, SSTR antibody, Smstr28 antibody, somatostatin receptor 3 antibody, SSTR3 antibody, LOC100303654 antibody, Sstr3 antibody

Background: 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