Key Product Details

Species Reactivity

Rabbit

Applications

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

Label

HRP

Antibody Source

Polyclonal Donkey IgG

Format

Pre-adsorbed

Concentration

0.5 mg/ml

View all IgG (H+L) Secondary Antibodies »

Product Specifications

Immunogen

This Donkey anti-Rabbit IgG (H+L) Secondary Antibody [HRP] (Pre-adsorbed) was developed against rabbit IgG-heavy and light chain.

Specificity

By immunoelectrophoresis and ELISA this Donkey anti-Rabbit IgG (H+L) Secondary Antibody [HRP] (Pre-adsorbed) reacts specifically with rabbit IgG and with light chains common to other rabbit immunoglobulins. No antibody was detected against non-immunoglobulin serum proteins. Antiserum was cross adsorbed using bovine, chicken, goat, human, mouse, pig and rat immunosorbents to remove cross reactive Antibodies. Less than 0.1% cross reactivity to bovine, chicken, goat, horse, human, mouse, pig and rat IgG was detected. This antibody may cross react with IgG from other species.

Clonality

Polyclonal

Host

Donkey

Isotype

IgG

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

Application
Recommended Usage

ELISA

1:10000-1:100000

Immunocytochemistry/ Immunofluorescence

1:200 - 1:5000

Immunohistochemistry

1:200 - 1:5000

Immunohistochemistry-Paraffin

1:200 - 1:5000

Simple Western

1:50

Western Blot

1:5000 - 1:50000
Application Notes
In Simple Western internal validation: Separated by size; antibody dilution of 1:50; matrix was 12-230 kDa; detected by Chemiluminescence.. See Simple Western Antibody Database for Simple Western validation: separated by size; antibody dilution of 1:50; matrix was 12-230 kDa; detected by Chemiluminescence.

Reviewed Applications

Read 1 review rated 5 using NB7185 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

Phosphate Buffered Saline (PBS) containing 0.2% BSA

Format

Pre-adsorbed

Preservative

0.05% Pro-Clean 400

Concentration

0.5 mg/ml

Shipping

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

Stability & Storage

Store at 4C. Do not freeze.

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-Rabbit IgG (H+L) Secondary Antibody [HRP] (Pre-adsorbed)

Product Datasheets

Certificate of Analysis

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

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

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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