Glucagon Antibody

• Catalog Number: orb1784557
• Category: Antibodies
• Description: Glucagon Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, IF, IHC, WB
• Reactivity: Human
• 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.
• Concentration: 1.18 mg/mL
• Dilution range: ELISA: 1:24,800-1:44,800, IHC: User Optimized, WB: User Optimized
• Form/Appearance: Liquid (sterile filtered)
• Purity: This affinity purified antibody is directed against human Glucagon. This product was affinity purified from monospecific antiserum by immunoaffinity purification. A BLAST analysis was used to suggest cross-reactivity with the antigen based on 100% homology with the immunizing sequence to human, chimpanzee, and bonobo.
• Conjugation: Unconjugated
• UniProt ID: P01275
• NCBI: NP_002045.1
• Storage: 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.
• Buffer/Preservatives: Preservative: 0.01% (w/v) Sodium Azide. Stabilizer: None. 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
• Alternative names: Rabbit Anti-Glucagon, Pro-glucagon, Glicentin, Gli
• Note: For research use only
• Application notes: Anti-Glucagon is tested in ELISA, IF, and Western Blot. Although not tested, this antibody is suitable in immunohistochemistry. 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
• Expiration Date: 12 months from date of receipt.

ELISA Results of Rabbit Anti-Glucagon Antibody. Each well was coated with 1 µg of conjugate. The starting concentration of antibody in the dilution series was 5 µg/ml. The titer is 1:34800 Glucagon - Free peptide [Green Line], 1:47200 Glucagon Standard - BSA conjugated [Blue Line], and 1:20500 Glucagon - BSA conjugated [Purple Line]. Each point on the Y-axis represents a 3-fold dilution. 3% Fish Gel (p/n orb348587), HRP conjugated Goat anti-Rabbit IgG (H&L) (p/n orb347673), and TMB substrate were used for detection.

Immunofluorescence of Rabbit Anti-Glucagon Antibody. Cell Line: NIH/3T3 cells. Fixative: 100% Methanol. Permeabilization: Triton X-100. Primary Antibody: Anti-Glucagon at 15 µg/mL overnight at 2-8°C. Secondary Antibody: Goat Anti-Rabbit IgG DyLight™488 at 5 µL/mL for 1 hr at RT. Nuclear Counterstain: DAPI. Staining: (A). DAPI. (B). Anti-Glucagon + DyLight™488 secondary. (C). Merge A+ B. (D). secondary only. Localization expected: Cytoplasm.

Immunofluorescence of Rabbit Anti-Glucagon Antibody. Cell Line: NIH/3T3 cells. Fixative: 4% PFA. Permeabilization: Triton X-100. Primary Antibody: Anti-Glucagon at 15 µg/mL overnight at 2-8°C. Secondary Antibody: Goat Anti-Rabbit IgG DyLight™488 at 5 µL/mL for 1 hr at RT. Nuclear Counterstain: DAPI. Staining: (A). DAPI. (B). Anti-Glucagon + DyLight™488 secondary. (C). Merge A+ B. (D). secondary only. Localization expected: Cytoplasm.

Western Blot of Rabbit Anti-Glucagon Antibody. Lane 1: Opal Prestained Molecular Weight. Lane 2: COS-7 Lysate - reduced (20 µg). Lane 3: BSA Conjugated Glucagon peptide - reduced (0.02 µg). Lane 4: Insulin - reduced (0.05 µg). Primary Antibody: Anti-Glucagon [Rabbit] Antibody at 1.0 µg/mL overnight at 2-8°C. Secondary Antibody: Goat Anti-Rabbit IgG (MX10) Peroxidase conjugated at 1:70000 for 30 mins at RT. Block: Blocking Buffer for Fluorescent Western Blotting (p/n orb348637) for 1 hr at RT. Expected MW: ~21 kDa. Observed MW: endogenous detection in COS-7 Lysate at ~35 kDa. Glucagon peptide is detected at the MW of BSA. No cross-reactivity with insulin is observed. Exposure: 25 sec.