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Donkey anti-Goat IgG (H+L) Secondary Antibody [HRP] (Pre-adsorbed)

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Product Details

Summary
Reactivity GtSpecies Glossary
Applications WB, ELISA, IHC
Clonality
Polyclonal
Host
Donkey
Conjugate
HRP
Format
Pre-adsorbed
Concentration
LYOPH

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Donkey anti-Goat IgG (H+L) Secondary Antibody [HRP] (Pre-adsorbed) Summary

Description
Store vial at 4C prior to restoration. For extended storage aliquot contents and freeze at -20C or below. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use.

This product was prepared from monospecific antiserum by immunoaffinity chromatography using Goat 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-Peroxidase, anti-Donkey Serum, Goat IgG and Goat Serum
Immunogen
Donkey anti-Goat IgG (H+L) Secondary Antibody [HRP] (Pre-adsorbed) was produced by repeated immunization with Goat IgG whole molecule in donkey.
Specificity
This antibody was pre-adsorbed against Chicken, Guinea Pig, Hamster, Horse, Mouse, Rabbit, and Rat Serum Proteins.

No reaction was observed against Chicken, Guinea Pig, Hamster, Horse, Mouse, Rabbit and Rat Serum Proteins.
Isotype
IgG
Clonality
Polyclonal
Host
Donkey
Purity
Multi-step

Applications/Dilutions

Dilutions
  • ELISA 1:170000
  • Immunohistochemistry 1:500 - 1:2500
  • Western Blot 1:2000 - 1:10000
Application Notes
This product has been tested by ELISA and is suitable for use in immunoelectrophoresis, western-blot, competitive western-blot, ELISA and competitive ELISA assays. Specific conditions for reactivity and signal detection should be optimized by the end user.

NOTE: Do NOT add Sodium Azide (it inhibits HRP irreversibly).

Packaging, Storage & Formulations

Storage
Store lyophilized antibody at 4C in the dark. Aliquot reconstituted liquid and store at -20C. Avoid freeze-thaw cycles.
Buffer
Lyophilized from 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 10 mg/mL Bovine Serum Albumin (BSA) - Immunoglobulin and Protease free
Preservative
0.01% Gentamicin Sulfate
Concentration
LYOPH
Purity
Multi-step
Reconstitution Instructions
Reconstitute with 1.0 ml deionized water (or equivalent).

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.

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