Key Product Details

Species Reactivity

Mouse

Applications

ELISA, Immunohistochemistry, Western Blot

Label

HRP

Antibody Source

Polyclonal Rabbit IgG

Format

Pre-adsorbed

Concentration

LYOPH mg/ml

View all IgG (H+L) Secondary Antibodies »

Product Summary for Rabbit anti-Mouse IgG (H+L) Secondary Antibody [HRP] (Pre-adsorbed)

Immunogen

Mouse IgG whole molecule

Specificity

This antibody was pre-adsorbed against Human Serum Proteins. No reaction was observed against Human Serum Proteins. Specificity was confirmed by ELISA at less than 1% cross reactivity against other mouse or human heavy or light chain isotypes.

Clonality

Polyclonal

Host

Rabbit

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 Mouse IgG coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Peroxidase, anti-Rabbit Serum, Mouse IgG, and Mouse Serum

Scientific Data Images for Rabbit anti-Mouse IgG (H+L) Secondary Antibody [HRP] (Pre-adsorbed)

Western Blot: Rabbit anti-Mouse IgG (H+L) Secondary Antibody [HRP] (Pre-adsorbed) [NBP1-73435] - WB shows detection of mouse anti-beta Actin monoclonal antibody (Catalog# NBP1-47423) using a HRP conjugated rabbit anti-mouse IgG H/L chains secondary antibody (Catalog# NBP1-73435). Image from verified customer review.
ELISA: Rabbit anti-Mouse IgG (H+L) Secondary Antibody [HRP] (Pre-adsorbed) [NBP1-73435] - ELISA results of purified Rabbit anti-Mouse IgG (H+L) Secondary Antibody [HRP] (Pre-adsorbed) tested against purified Mouse IgG. (Red Line). Each well was coated in duplicate with 1.0 ug of antigen. 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 Blocking buffer

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

Application
Recommended Usage

ELISA

1:150000

Immunohistochemistry

1:200 - 1:1000

Western Blot

1:500 - 1:2500
Application Notes
This product has been tested by ELISA and is suitable for ELISA, Sandwich ELISA, titration assays, western-blot, immunoprecipitation, Immunohistochemistry. Specific conditions for reactivity and signal detection should be optimized by the end user.

NOTE: Do NOT add Sodium Azide (it inhibits HRP irreversibly).
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 1 review rated 5 using NBP1-73435 in the following applications:

Formulation, Preparation, and Storage

Purification

Multi-step

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

Format

Pre-adsorbed

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.

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 Rabbit anti-Mouse 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 Rabbit anti-Mouse 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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