Donkey anti-Mouse IgG (H+L) Secondary Antibody [PE/Cy5.5]

• Reactivity: Mu
• Applications: WB, ELISA, FLISA, IHC, IHC-Fr, IHC-P
• Clonality: Polyclonal
• Host: Donkey
• Conjugate: PE/Cy5.5

Donkey anti-Mouse IgG (H+L) Secondary Antibody [PE/Cy5.5] Summary

• Immunogen: This Donkey anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) was developed against mouse IgG whole molecule.
• Specificity: Assay of Donkey anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) by immunoelectrophoresis resulted in a single precipitin arc against anti-Donkey serum, Mouse IgG and Mouse serum. No reaction was observed against Bovine, Chicken, Goat, Guinea Pig, Hamster, Horse, Human, Rabbit, Rat and Sheep serum proteins. Specificity was confirmed using ELISA at less than 1.0% cross reactivity against IgG proteins purified from the species listed above. This antibody was pre-adsorbed against bovine chicken goat guinea pig hamster horse human rabbit rat & sheep serum proteins.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Donkey
• Purity: Immunogen affinity purified

Applications/Dilutions

• Application Notes: Optimal dilution of this antibody should be experimentally determined. For optimal results using our Tandem dyes, please avoid prolonged exposure to light or extreme temperature fluctuations. These can lead to irreversible degradation or decoupling. When staining intracellular targets, specific attention to the fixation and permeabilization steps in your flow protocol may be required. Please contact our technical support team at technical@novusbio.com if you have any questions.

Packaging, Storage & Formulations

• Storage: Store at 4C in the dark. Do not freeze.
• Buffer: PBS
• Preservative: 0.05% Sodium Azide
• Purity: Immunogen affinity purified

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.