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Key Product Details

Species Reactivity

Rabbit

Applications

ELISA, Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot

Label

Alkaline Phosphatase

Antibody Source

Polyclonal Goat IgG

Format

Pre-adsorbed

Concentration

1.5 mg/ml

Product Specifications

Immunogen

This Goat anti-Rabbit IgG (H+L) Secondary Antibody [Alkaline Phosphatase] (Pre-adsorbed) was developed against purified rabbit IgG (H&L).

Reactivity Notes

Based on IEP, no reactivity is observed to non-immunoglobulin rabbit serum proteins, serum proteins from bovine, human, or mouse, and IgG from human or mouse

Specificity

Based on IEP, this Goat anti-Rabbit IgG (H+L) Secondary Antibody [Alkaline Phosphatase] (Pre-adsorbed) heavy gamma chains on rabbit IgG and light chains on all rabbit immunoglobulins. This antibody has been pre-adsorbed against bovine, human, or mouse IgG

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Description

Purity > 95% based on SDS-PAGE. Goat serum was obtained from healthy animals of US origin and under the care of a registered veterinarian.

Store undiluted liquid at 2-8C. For storage at -20C, dilute with an equal volume of glycerol to prevent loss of enzymatic activity. Prepare working dilutions prior to use and then discard.

Applications

Application
Recommended Usage

ELISA

1:200-1:5000

Immunohistochemistry

1:500-1:5000

Western Blot

1:200-1:5000
Application Notes
This conjugate is suitable for all immunoassay applications.

Formulation, Preparation, and Storage

Purification

Affinity purified

Formulation

30 mM Triethanolamine, pH 7.2, 5 mM Magnesium Chloride, 0.1 mM Zinc Chloride, 1 % (w/v) BSA, Protease/IgG free

Format

Pre-adsorbed

Preservative

0.05% Sodium Azide

Concentration

1.5 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. After dilution, add an equal volume of glycerol to prevent loss of enzymatic activity and store at -20C. Avoid repeated freeze-thaw cycles.

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

Certificate of Analysis

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

Product Specific Notices

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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