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

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

• Immunogen: Chickens were immunized with purified, non-denatured goat IgG (containing both heavy and light chains).
• Specificity: There is no detectable cross-reactivity with rabbit, human or mouse IgG.
• Isotype: IgY
• Clonality: Polyclonal
• Host: Chicken
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • ELISA
  • Western Blot 1:5000
• Application Notes: Antibodies were tested for specificity using double- immunodiffusion in agarose gels and ELISA analysis. In the latter, a 1:75000 dilution gave an O.D. of 1.0 in a 15 minute reaction using tetramethylbenzidine as substrate.
• Readout System:
  • NovaLume Plus Chemiluminescent Reagent LUM-555

Reactivity Notes

There is no detectable cross-reactivity with rabbit, human or mouse IgG.

Packaging, Storage & Formulations

• Storage: Store at 4C in the dark.
• Buffer: HEPES (50 mM, pH 7.0), NaCl (0.1 M), bovine serum albumin (0.2%, w/v).
• Preservative: 0.01% Thimerosal
• Concentration: 1.0 mg/ml
• Purity: Immunogen affinity purified

Notes

Purification Notes
Chickens were immunized with purified, non-denatured goat IgG (containing both heavy and light chains). Eggs were collected from hyperimmunized hens and the IgY fraction purified from the yolks. Anti-goat IgGs were affinity-purified over a column conjugated with purified goat IgG (containing both heavy and light chains) and then conjugated with horseradish peroxidase.

Storage Notes
Store at 4C in the dark. Under these conditions, the antibodies should have a shelf life of at least 12 months (provided they remain sterile). For longer storage periods, dilute the antibodies with sterile glycerol and store at -20C. Do not freeze these antibodies unless you want to store them for longer periods of time.

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.

⚠ WARNING: This product can expose you to chemicals including mercury, which is known to the State of California to cause reproductive toxicity with developmental effects.  For more information go to www.P65Warnings.ca.gov.