Key Product Details

Species Reactivity

Goat

Applications

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, ELISA, Immunocytochemistry/ Immunofluorescence

Label

HRP

Antibody Source

Polyclonal Donkey IgG
View all IgG (H+L) Secondary Antibodies »

Product Specifications

Immunogen

Donkey anti-Goat IgG (H+L) Secondary Antibody [HRP] was produced by repeated immunization with Goat IgG whole molecule in donkey.

Clonality

Polyclonal

Host

Donkey

Isotype

IgG

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. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Peroxidase, anti-Donkey Serum, Goat IgG and Goat Serum.

Scientific Data Images for Donkey anti-Goat IgG (H+L) Secondary Antibody [HRP]

ELISA: Donkey anti-Goat IgG (H+L) Secondary Antibody [HRP] [NBP1-73223] - ELISA results of purified Donkey anti-Goat IgG (H+L) Secondary Antibody [HRP] tested against purified Goat IgG. Each well was coated in duplicate with 1.0 ug of Goat IgG

Applications for Donkey anti-Goat IgG (H+L) Secondary Antibody [HRP]

Application
Recommended Usage

ELISA

1:10000 - 1:150000

Immunocytochemistry/ Immunofluorescence

0.25-2 ug/ml

Immunohistochemistry

1:500 - 1:2500

Immunohistochemistry-Paraffin

1:200-1:500

Western Blot

1:1000 - 1:5000
Application Notes
This product is suitable for use in immunoelectrophoresis, western-blot, competitive western-blot, ELISA and competitive ELISA assays. It is also suitable in IHC. Specific conditions for reactivity and signal detection should be optimized by the end user.

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

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Reconstitution

Reconstitute with 1.0 ml deionized water (or equivalent).

Formulation

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 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store lyophilized antibody at 4C in the dark. Aliquot reconstituted liquid and store at -20C. Avoid freeze-thaw cycles.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: IgG (H+L)

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

Additional IgG (H+L) Products

Product Documents for Donkey anti-Goat IgG (H+L) Secondary Antibody [HRP]

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Product Specific Notices for Donkey anti-Goat IgG (H+L) Secondary Antibody [HRP]

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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Protocols

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FAQs

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