Glucagon antibody

Catalog No. ABIN7448121

Supplier Product No.: 600-401-mv9

This independently validated Rabbit Polyclonal antibody specifically detects Glucagon in WB, IHC, ELISA, IF and FM. It exhibits reactivity toward Human.

Quick Overview for Glucagon antibody (ABIN7448121)

Target: Glucagon (GCG)

Reactivity: Human

Host: Rabbit

Clonality: Polyclonal

Conjugate: This Glucagon antibody is un-conjugated

Application: Western Blotting (WB), Immunohistochemistry (IHC), ELISA, Immunofluorescence (IF), Fluorescence Microscopy (FM)

Product Details anti-Glucagon Antibody

Supplier Product No.: 600-401-mv9

Supplier: Rockland

Purpose: Glucagon Antibody

Cross-Reactivity (Details): This affinity purified antibody is directed against human Glucagon.

Characteristics: Synonyms: Rabbit Anti-Glucagon, Pro-glucagon, Glicentin, Oxyntomodulin, OXM, OXY, Glucagon

Purification: This product was affinity purified from monospecific antiserum by immunoaffinity purification.

Sterility: Sterile filtered

Immunogen:

Immunogen: Glucagon antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to an internal portion of human Glucagon.

Immunogen Type: Conjugated Peptide

Isotype: IgG

Application Details

Application Notes:

Immunohistochemistry Dilution: 1:100

Application Note: Anti-Glucagon is tested in ELISA, IF, IHC, and Western Blot. Expect a band approximately ~20.9 kDa corresponding to the appropriate cell lysate or extract. Specific conditions for reactivity should be optimized by the end user.

Western Blot Dilution: User Optimized

ELISA Dilution: 1:24,800-1:44,800

IF Microscopy Dilution: 5-15 μg/mL

Restrictions: For Research Use only

Independent Validation (IHC)

Validation #104519 (Immunohistochemistry)

by: Prof. Merighi, Laboratory of Neurobiology, Department of Veterinary Sciences, University of Turin

No.: #104519

Date: 01/26/2024

Antigen: Glucagon

Lot Number: 49549

Method validated: Immunohistochemistry

Positive Control:

Adult (> 2 months) CD1 mouse pancreas fixed in 4% paraformaldehyde.

Negative Control:

One control slice for each experimental procedure processed omitting the primary antibody; overnight incubation in diluent solution only.

Notes:

Passed. The Glucagon antibody ABIN7448121 works in IHC-P at 1:100 concentrations with microwave antigen retrieval treated sections.

Validation Images

Staining with ABIN7448121 of glucagon immunostained α cells in the CD1 mouse pancreatic islets. Original magnification 20x.

Staining with ABIN7448121 of glucagon immunostained α cells in the CD1 mouse pancreatic islets. Original magnification 20x.

Full Methods

Primary Antibody: ABIN7448121

Secondary Antibody: anti-rabbit secondary antibody Alexa Fluor 488 (Thermo Fisher Scientific, A11034, lot # XJ357262)

Full Protocol:

• Perfuse mice with paraformaldehyde 4% in 0.1 M phosphate buffer pH 7.4 and post-fix of samples in the same fixative for an additional 2 h at RT.
• Wash, dehydrate, and embed samples in paraffin wax.
• Wash several times with 0.01 M PBS.
• Cut brain with a microtome into 6 µm sections and mount on glass slides.
• After paraffin removal processed sections for microwave antigen retrieval for 10 minutes (95-100° C) in 10 mM sodium citrate buffer (pH 6.0). After 20 minutes of spontaneous cooling, wash sections (distilled water, 2 times for 5 minutes each; followed by PBS for 5 minutes).
• Incubate sections for 1 h at RT in a humid chamber in PBS containing 1% ovalbumin (Sigma, A5378) and 0.3% Triton-X-100 (BioRad, 161-0407, lot 00583) to block non-specific binding sites.
• Incubate sections with primary rabbit anti- Glucagon (antibodies-online, ABIN7448121, lot 49549) diluted 1:50 and 1:100, in PBS-BSA-PLL ON at RT in a humid chamber.
• Wash sections 3x 5 min with 0.01 M PBS.
• Incubate sections with anti-rabbit secondary antibody Alexa Fluor 488 (Thermo Fisher Scientific, A11034, lot # XJ357262), 1:500, in 0.1M PBS, 1 hour at room temperature.
• Wash sections 3x 5 min in 0.01 M PBS.
• Mount specimens in Fluoroshield (Sigma, F6182, lot MKCB0153V).
• Acquire images Leica DM 6000B fluorescence microscope equipped with a digital camera at 40x magnification.

Handling

Format: Liquid

Concentration: 1.18 mg/mL

Buffer:

Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2

Stabilizer: None

Preservative: 0.01 % (w/v) Sodium Azide

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: 4 °C,-20 °C

Storage Comment: Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.

Expiry Date: 12 months

Target Details for Glucagon

Target: Glucagon (GCG)

Alternative Name: GCG

Background: Background: Glucagon is a member of a multigene family that includes secretin. Glucagon is a 29-amino acid pancreatic hormone that counteracts the glucose-lowering action of insulin by stimulating glycogenolysis and gluconeogenesis. The human glucagon gene is approximately 9.4 kb long, contains 6 exons and 5 introns, and assigned to 2q36-2q37. This antibody is suitable for researchers interested in metabolic diseases, like diabetes, cell proliferation, differentiation, apoptosis, GCPR signaling, and calcium signaling research.

Gene ID: 2641

NCBI Accession: NP_002045

UniProt: P01275

Pathways: Positive Regulation of Peptide Hormone Secretion, Peptide Hormone Metabolism, cAMP Metabolic Process, Regulation of Carbohydrate Metabolic Process, Feeding Behaviour, Negative Regulation of intrinsic apoptotic Signaling