Glucagon Antibody

SKU: orb1784557

Description

Glucagon Antibody

Images & Validation

• Tested Applications: ELISA, IF, IHC, WB
• Dilution range: ELISA: 1:24,800-1:44,800, IHC: 1:100, IF: 5-15µg/mL
• Reactivity: Human
• Application Notes:
  Anti-Glucagon is tested in ELISA, IF, IHC-P, 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

Key Properties

• Antibody Type: Primary Antibody
• Host: Rabbit
• Clonality: Polyclonal
• 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.
• 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

Storage & Handling

• 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.
• Form/Appearance: Liquid (sterile filtered)
• Buffer/Preservatives: Preservative: 0.01% (w/v) Sodium Azide. Stabilizer: None; Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
• Concentration: 1.18 mg/mL
• Dry Ice Shipping: Please note: This product requires shipment on dry ice. A dry ice surcharge will apply.
• Disclaimer: For research use only

Alternative Names

Rabbit Anti-Glucagon, Pro-glucagon, Glicentin, Oxyntomodulin, OXM, OXY, Glucagon

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: MCF7 cells. Fixative: 100% Methanol. Permeabilization: 0.3% 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 1hr 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: 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 1hr 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 1hr 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: U20S cells. Fixative: 4% PFA. Permeabilization: 0.3% 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 1hr at RT. Nuclear Counterstain: DAPI. Staining: (A). DAPI. (B). Anti-Glucagon + DyLight™488 secondary. (C). Merge A + B. (D). secondary only. Localization expected: Cytoplasm.

Immunohistochemistry results using Rabbit Anti-Glucagon Antibody. Tissue: alpha cells in CD1 mouse pancreatic islets. Fixation: 4% paraformaldehyde. Antigen Retrieval: 10 mM Sodium Citrate buffer for 10 mins at 95-100°C. Blocking: PBS, 1% ovalbumin, 0.3% Triton X-100. Primary Antibody: Anti-Glucagon at 1:100 overnight at RT. Secondary Antibody: Anti-Rabbit Alexa Fluor 488 at 1:500 for 1hr at RT. Original magnification 20x.

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 1hr at RT. Expected MW: ~21kDa. Observed MW: endogenous detection in COS-7 Lysate at ~35kDa. Glucagon peptide is detected at the MW of BSA. No cross-reactivity with insulin is observed. Exposure: 25 sec.

Quick Database Links

UniProt

P01275

RefSeq (Protein):NP_002045.1