Key Product Details

Species Reactivity

Rat

Applications

Immunohistochemistry, Western Blot, ELISA

Label

HRP

Antibody Source

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

Product Specifications

Immunogen

Rat IgG whole molecule

Clonality

Polyclonal

Host

Chicken

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 Rat IgG coupled to agarose. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Peroxidase, anti-Chicken Serum, Rat IgG and Rat Serum.

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

Western Blot: Chicken anti-Rat IgG (H+L) Secondary Antibody [HRP] [NB120-6836] - Western Blot of Chicken anti-Rat IgG (H+L) Secondary antibody [HRP]. Lane M: 3 ul Molecular Ladder. Lane 1: Rat IgG whole molecule. Lane 2: Rat IgG F(c) Fragment. Lane 3: Rat IgG Fab Fragment. Lane 4: Rat IgM Whole Molecule. Lane 5: Rat Serum. All samples were reduced. Load: 50 ng per lane. Block for 30 min at RT. Primary Antibody: Chicken anti-Rat IgG (H+L) Secondary antibody [HRP] 1:1,000 for 60 min at RT. Secondary Antibody: Anti-Chicken IgG (GOAT) Peroxidase Conjugated Antibody 1:40,000 in blocking buffer for 30 min at RT. Predicted/Obsevered Size: 25 and 55 kDa for Rat IgG and Serum, 25 kDa for F(c) and Fab, 78 and 25 kDa for IgM. Rat F(c) migrates slightly higher.

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

Application
Recommended Usage

ELISA

1:10000 - 1:100000

Immunohistochemistry

1:500 - 1:2500

Western Blot

1:1000 - 1:10000
Application Notes
This product has been tested by ELISA and is assayed against 1.0 ug of Rat IgG in a standard capture ELISA using ABTS (2,2'-azino-bis-[3-ethylbenthiazoline-6-sulfonic acid] substrate for 30 minutes at room temperature. A working dilution of 1:2,000 to 1:12,000 of the reconstitution concentration is suggested for this product. Optimal concentration in other immunoassays should be determined by researcher.

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 Chicken anti-Rat 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 Chicken anti-Rat IgG (H+L) Secondary Antibody [HRP]

Chicken products cannot be exported to Canada.

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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