Goat anti-Chicken IgG (H+L) Secondary Antibody

Western Blot: Goat anti-Chicken IgG (H+L) Secondary Antibody [NBP1-72720] - Lane 1: Chicken IgG whole molecule. Lane 2: Chicken IgG F(c) Fragment. Lane 3: Chicken IgG Fab Fragment. All samples were reduced. Load: 50 ng per lane. Block: incubated with blocking buffer for 30 min at RT. Primary Antibody: Anti-Chicken IgG (H&L) (GOAT) Antibody 1:1,000 for 60 min at RT. Secondary antibody: Anti-Goat IgG (DONKEY) Peroxidase Conjugated Antibody 1:40,000 in incubated with blocking buffer for 30 min at RT. Predicted/Observed Size: 25 and 72 kDa for Chicken IgG, 25 kDa for F(c) and Fab. Chicken F(c) migrates slightly higher. Image from the Alkaline Phosphatase version of this antibody.

ELISA: Goat anti-Chicken IgG (H+L) Secondary Antibody [NBP1-72720] - ELISA results of purified Goat anti-Chicken IgG (H+L) Secondary Antibody [Biotin] tested against purified Chicken IgG. Each well was coated in duplicate with 1.0 ug of Chicken IgG. The starting dilution of antibody was 5ug/ml and the X-axis represents the Log10 of a 3-fold dilution. This titration is a 4-parameter curve fit where the IC50 is defined as the titer of the antibody. Assay performed using blocking buffer, Streptavidin HRP conjugate, and TMB substrate.. Using the Biotin format of this antibody.

ELISA: Goat anti-Chicken IgG (H+L) Secondary Antibody [NBP1-72720] - ELISA results of purified Goat anti-Chicken IgG (H+L) Secondary Antibody [HRP] tested against purified Chicken IgG. Each well was coated in duplicate with 1.0 ug of Chicken IgG (red line). The starting dilution of antibody was 5ug/ml and the X-axis represents the Log10 of a 3-fold dilution. This titration is a 4-parameter curve fit where the IC50 is defined as the titer of the antibody. Assay performed using TMB substrate.. Using the HRP format of this antibody.

ELISA: Goat anti-Chicken IgG (H+L) Secondary Antibody [NBP1-72720] - ELISA Results of Goat anti-Chicken IgG (H+L) Secondary antibody tested against purified Chicken IgG/IgY. Each well was coated in duplicate with 1.0 ug of Chicken IgG/IgY. The working dilution is 1:47,000. The starting dilution of antibody was 5ug/ml and the X-axis represents the Log10 of a 3-fold dilution. This titration is a 4-parameter curve fit where the IC50 is defined as the titer of the antibody. Assay performed using HRP Conjugated Stabilizer, Donkey Anti-Goat IgG mx10 HRP conjugated, and TMB substrate.

• Reactivity: Ch
• Applications: WB, DB, ELISA, IHC
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated

Goat anti-Chicken IgG (H+L) Secondary Antibody Summary

• Description: This product was prepared from monospecific antiserum by immunoaffinity chromatography using Chicken IgG coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Goat Serum, Chicken IgG and Chicken Serum.
  
  Store vial at 4C prior to opening. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use. For extended storage aliquot contents and freeze at -20C or below. Avoid cycles of freezing and thawing.
• Immunogen: Chicken IgG whole molecule
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Purity: Multi-step

Applications/Dilutions

• Dilutions:
  • Dot Blot
  • ELISA 1:20000 - 1:100000
  • Immunohistochemistry 1:1000 - 1:5000
  • Western Blot 1:2000 - 1:10000
• Application Notes: This product has been tested by ELISA and is suitable for western blot and immunohistochemistry, as well as other assays requiring lot-to-lot consistency.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
• Preservative: 0.01% Sodium Azide
• Purity: Multi-step

Background

Antibodies, also known as immunoglobulins (Igs) are critical for immunity and are grouped into five primary classes: IgG, IgM, IgA, IgD, and IgE. The most abundant antibody isotype is immunoglobulin G (IgG) with concentrations ranging from 7.5-22 mg/ml in human serum and has a molecular weight of 150 kDa. The major effector functions of IgG include neutralization, opsonization, complement fixation and antibody dependent cell-mediated cytotoxicity (ADCC). This monomeric immunoglobulin, expressed on the surface of mature B cells, is often depicted as a Y-shape and comprised of 2 heavy chains and 2 light chains linked by disulfide bonds. The heavy chain is type gamma including subtypes gamma 1, gamma 2, gamma 3, and gamma 4 while the light chain is either a kappa or lambda chain. An IgG molecule has two antigen binding sites, each consisting of a heavy and light chain N-terminal variable domain. When combined with the constant heavy chain 1 (Ch1) and the constant light chain domains, it forms the fragment antigen-binding (Fab) region (2 per antibody). The remaining domains (Ch2-Ch4) of both heavy chains make up the Fc region and contain a site for covalently linking an enzymatic or fluorochrome probe, such as HRP or Janelia Fluor 549, for target detection and visualization (1,2,3). The 4 IgG subclasses, sharing 95% amino acid identity, include IgG1, IgG2, IgG3, and IgG4 for humans and IgG1, IgG2a, IgG2b, and IgG3 for mice. The relative abundance of each human subclass is 60% for IgG1, 32% for IgG2, 4% for IgG3, and 4% for IgG4. In an IgG deficiency, there may be a shortage of one or more subclasses (4). References 1. Painter RH. (1998) Encyclopedia of Immunology (Second Edition). Elsevier. 1208-1211 2. Chapter 9 - Antibodies. (2012) Immunology for Pharmacy. Mosby 70-78 3. Schroeder H, Cavacini, L. (2010) Structure and Function of Immunoglobulins. J Allergy Clin Immunol. 125(2 0 2): S41-S52. PMID: 20176268 4. Vidarsson G, Dekkers G, Rispens T. (2014) IgG subclasses and allotypes: from structure to effector functions. Front Immunol. 5:520. PMID: 25368619

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Secondary Antibodies are guaranteed for 1 year from date of receipt.

Publications for IgG (H+L) Antibody (NBP1-72720)(1)

Publication using NBP1-72720

• Amer SA, Al-Khalaifah HS, Gouda A et al. Potential Effects of Anthocyanin-Rich Roselle (Hibiscus sabdariffa L.) Extract on the Growth, Intestinal Histomorphology, Blood Biochemical Parameters, and the Immune Status of Broiler Chickens Antioxidants (Basel, Switzerland) 2022-03-13 [PMID: 35326194] (IF/IHC)