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Donkey anti-Mouse IgG (H+L) Secondary Antibody

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Western Blot: Donkey anti-Mouse IgG (H+L) Secondary Antibody [NB120-6707] - Lane 1: Mouse IgG. Lane 2: None. Load: 50 ng per lane. Primary antibody: None. Secondary antibody: Fluorescein donkey secondary antibody at ...read more
Immunocytochemistry/ Immunofluorescence: Donkey anti-Mouse IgG (H+L) Secondary Antibody [NB120-6707] - Immunocytochemistry/ Immunofluorescence: Donkey anti-Mouse IgG (H+L) Secondary Antibody [DyLight 488] (Pre-absorbed) ...read more
Western Blot: Donkey anti-Mouse IgG (H+L) Secondary Antibody [NB120-6707] - Lane 1: Mouse IgG. Lane 2: None. Load: 50 ng per lane. Primary Antibody: None. Secondary antibody: Mouse IgG (H&L) Antibody Peroxidase ...read more
Western Blot: Donkey anti-Mouse IgG (H+L) Secondary Antibody [NB120-6707] - Lane 1: Mouse IgG. Lane 2: None. Load: 50 ng per lane. Primary antibody: None. Secondary antibody: Texas Red donkey secondary antibody at ...read more
Western Blot: Donkey anti-Mouse IgG (H+L) Secondary Antibody [NB120-6707] - DyLight (R) dyes can be used for two-color western blot detection with low background and high signal. Image using the DyLight 680 form of this ...read more
Fluorophore-linked immunosorbent assay: Donkey anti-Mouse IgG (H+L) Secondary Antibody [NB120-6707] - Using the DyLight 405 format of this antibody.
ELISA: Donkey anti-Mouse IgG (H+L) Secondary Antibody [NB120-6707] - ELISA results of purified Donkey anti-Mouse IgG (H+L) Secondary Antibody [Biotin] tested against Mouse IgG. Each well was coated in duplicate with 1.0 ...read more
ELISA: Donkey anti-Mouse IgG (H+L) Secondary Antibody [NB120-6707] - ELISA Results of Donkey anti-Mouse IgG (H+L) Secondary antibody tested against purified Mouse IgG. Each well was coated in duplicate with 1.0 ug of ...read more

Product Details

Summary
Reactivity MuSpecies Glossary
Applications WB, ELISA, ICC/IF, IHC
Clonality
Polyclonal
Host
Donkey
Conjugate
Unconjugated

Donkey anti-Mouse IgG (H+L) Secondary Antibody Summary

Description
This product was prepared from monospecific antiserum by immunoaffinity chromatography using Mouse IgG coupled to agarose. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Donkey Serum, Mouse IgG and Mouse 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
Mouse IgG whole molecule
Isotype
IgG
Clonality
Polyclonal
Host
Donkey
Purity
Immunogen affinity purified

Applications/Dilutions

Dilutions
  • ELISA 1:50000 - 1:150000
  • Fluorophore-linked immunosorbent assay
  • Immunocytochemistry/ Immunofluorescence
  • Immunohistochemistry 1:1000 - 1:5000
  • Western Blot 1:2000 - 1:10000
Application Notes
This product has been tested by ELISA and is suitable for ELISA, western blot, and immunohistochemistry, as well as other assays requiring lot-to-lot consistency.
Publications
Read Publication using NB120-6707.

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
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.

Publications for IgG (H+L) Antibody (NB120-6707)(1)

We have publications tested in 1 application: WB.


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Product General Protocols

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Video Protocols

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ICC/IF Video Protocol

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