Donkey anti-Mouse IgG (H+L) Secondary Antibody [DyLight 650]

• Reactivity: Mu
• Applications: WB, Flow, IHC
• Clonality: Polyclonal
• Host: Donkey
• Conjugate: DyLight 650
• Concentration: LYOPH

Donkey anti-Mouse IgG (H+L) Secondary Antibody [DyLight 650] Summary

• Description: Affinity purified antibody is > 95% based on SDS-PAGE. Donkey serum was obtained from healthy animals of US origin and under the care of a registered veterinarian. DyLight 650 (Ex = 652 nm; Em = 672 nm). After reconstituted, prepare fresh working dilution daily
  
  Product is stable for up to 4 weeks at 2-8C after rehydration. For extended storage after rehydration, add an equal volume of glycerol and store at -20C.
• Immunogen: This Donkey anti-Mouse IgG (H+L) Secondary Antibody [DyLight 650] was developed against purified mouse IgG (H&L).
• Specificity: Based on IEP, this Donkey anti-Mouse IgG (H+L) Secondary Antibody [DyLight 650] reacts with heavy gamma chains on mouse IgG and light chains on all mouse immunoglobulins.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Donkey
• Purity: Affinity purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 1:20 - 1:2000
  • Immunomicroscopy 1:20 - 1:2000
  • Western Blot
• Application Notes: This conjugate is suitable for immunomicroscopy and flow cytometry.
  
  Use in Western Blot was reported in the scientific literature (PMID: 31300552).

Reactivity Notes

Based on IEP, no reactivity is observed to non-immunoglobulin mouse serum immunoglobulins

Packaging, Storage & Formulations

• Storage: Store lyophilized antibody at 4C. Mix reconstituted liquid with an equal volume of glycerol, aliquot contents and freeze at -20C or below for long term storage.
• Buffer: 10 mM Sodium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 1 % (w/v) BSA, Protease/IgG free
• Preservative: 0.05% Sodium Azide
• Concentration: LYOPH
• Purity: Affinity purified
• Reconstitution Instructions: Rehydrate with 1.1 ml of deionized water and let stand 30 minutes at room temperature to dissolve. (Product has been overfilled to ensure complete recovery.) Centrifuge to remove any particulates. Prepare fresh working dilution daily.

Notes

DyLight (R) is a trademark of Thermo Fisher Scientific Inc. and its subsidiaries.

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 (NBP1-75614)(1)

Publication using NBP1-75614

• LaRock DL, Sands JS, Ettouati E et al. Inflammasome inhibition blocks cardiac glycoside cell toxicity J Biol Chem 2019-07-14 [PMID: 31300552] (WB)