Key Product Details

Species Reactivity

Chicken

Applications

ELISA, Immunohistochemistry, Western Blot

Label

HRP

Antibody Source

Polyclonal Goat IgG

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

View all IgG (H+L) Secondary Antibodies »

Product Specifications

Immunogen

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

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Description

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

This product was prepared from monospecific antiserum by immunoaffinity chromatography using Chicken IgG coupled to agarose beads. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Peroxidase, anti-Goat Serum, Chicken IgG and Chicken Serum.

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

Western Blot: Goat anti-Chicken IgG (H+L) Secondary Antibody [HRP] [NB7608] - Goat anti-Chicken IgG (H+L) Secondary Antibody [HRP] [NB7608] - Lane 1: Chicken IgG whole molecule. Lane 2: Chicken IgG F(c) Fragment. Lane 3: Chicken IgG Fab Fragment. All samples were reduced. Load: 50 ng per lane. Block: incubated with blocking buffer for 30 min at RT. Primary Antibody: Anti-Chicken IgG (H&L) (GOAT) Antibody 1:1,000 for 60 min at RT. Secondary antibody: Anti-Goat IgG (DONKEY) Peroxidase Conjugated Antibody 1:40,000 in incubated with blocking buffer for 30 min at RT. Predicted/Observed Size: 25 and 72 kDa for Chicken IgG, 25 kDa for F(c) and Fab. Chicken F(c) migrates slightly higher.
ELISA: Goat anti-Chicken IgG (H+L) Secondary Antibody [HRP] [NB7608] - ELISA results of purified Goat anti-Chicken IgG (H+L) Secondary Antibody [HRP] tested against purified Chicken IgG. Each well was coated in duplicate with 1.0 ug of Chicken IgG (red line). The starting dilution of antibody was 5ug/ml and the X-axis represents the Log10 of a 3-fold dilution. This titration is a 4-parameter curve fit where the IC50 is defined as the titer of the antibody. Assay performed using TMB substrate.

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

Application
Recommended Usage

ELISA

1:20000 - 1:100000

Immunohistochemistry

1:1000 - 1:10000

Western Blot

1:1000 - 1:5000
Application Notes
This product is tested in immunoelectrophoresis, western-blot, competitive western-blot, ELISA and competitive ELISA assays. Anti-Chicken IgG HRP Antibody is suitable for 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

Formulation

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

Please see the vial label for concentration. If unlisted please contact technical services.

Shipping

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

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid 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 for Goat anti-Chicken IgG (H+L) Secondary Antibody [HRP]

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 Goat anti-Chicken 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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