Key Product Details

Species Reactivity

Human

Applications

ELISA, Immunohistochemistry, Peptide ELISA, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit 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

Human IgG whole molecule

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Description

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

Store vial at 4C prior to opening. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use. For extended storage aliquot contents and freeze at -20C or below. Avoid cycles of freezing and thawing.

Scientific Data Images for Rabbit anti-Human IgG (H+L) Secondary Antibody

Western Blot: Rabbit anti-Human IgG (H+L) Secondary Antibody [NB120-6715]

Western Blot: Rabbit anti-Human IgG (H+L) Secondary Antibody [NB120-6715]

Western Blot: Rabbit anti-Human IgG (H+L) Secondary Antibody [NB120-6715] - Lane 1: Human IgG. Lane 2: None. Load: 50 ng per lane. Primary antibody: Human IgG Antibody biotin conjugated at 1:1,000 for 60 min at RT. Secondary antibody: Peroxidase streptavidin secondary antibody at 1:40,000 for 30 min at RT. Blocking: incubated with blocking buffer for 30 min at RT. Predicted/Observed size: 25 & 55 kDa, 25 & 55 kDa for Human IgG. Other band(s): None.
Western Blot: Rabbit anti-Human IgG (H+L) Secondary Antibody [NB120-6715]

Western Blot: Rabbit anti-Human IgG (H+L) Secondary Antibody [NB120-6715]

Western Blot: Rabbit anti-Human IgG (H+L) Secondary Antibody [NB120-6715] - Lane 1: Human IgG Load: 50 ng per lane Primary antibody: Human IgG (H&L) Antibody Biotin Conjugated at 1:1000 for 60 min RT Secondary antibody: HRP Conjugated Streptavidin at 1:40,000 for 30 min at RT Block: incubated with blocking buffer for 30 min at RT
ELISA: Rabbit anti-Human IgG (H+L) Secondary Antibody [NB120-6715]

ELISA: Rabbit anti-Human IgG (H+L) Secondary Antibody [NB120-6715]

ELISA: Rabbit anti-Human IgG (H+L) Secondary Antibody [NB120-6715] - ELISA results of purified Rabbit anti-Human IgG (H+L) Secondary Antibody [HRP] (Pre-adsorbed) tested against purified Human IgG. Each well was coated in duplicate with 1.0 ug of Human 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.. Using the HRP format of this antibody.

Applications for Rabbit anti-Human IgG (H+L) Secondary Antibody

Application
Recommended Usage

ELISA

1:1000000

Immunohistochemistry

1:2000 - 1:10000

Western Blot

1:10000 - 1:50000
Application Notes
This product has been tested by ELISA and is suitable for western blot and immunohistochemistry, as well as other assays requiring lot-to-lot consistency.

Use in Peptide ELISA was reported in the scientific literature (PMID: 22720696).

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2

Preservative

0.01% Sodium Azide

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 Rabbit anti-Human IgG (H+L) Secondary Antibody

Product Datasheets

Certificate of Analysis

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

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Product Specific Notices for Rabbit anti-Human IgG (H+L) Secondary Antibody

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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