Key Product Details

Species Reactivity

Rabbit

Applications

ELISA, Immunohistochemistry, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Guinea Pig IgG

Format

Pre-adsorbed

Concentration

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

View all IgG (H+L) Secondary Antibodies »

Product Summary for Guinea Pig anti-Rabbit IgG (H+L) Secondary Antibody (Pre-adsorbed)

Immunogen

Rabbit IgG whole molecule

Specificity

This antibody was pre-adsorbed against Human, Goat, and Mouse Serum Proteins. No reaction was observed against Goat, Human and Mouse Serum Proteins.

Clonality

Polyclonal

Host

Guinea Pig

Isotype

IgG

Description

This product was prepared from monospecific antiserum by immunoaffinity chromatography using Rabbit 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-Guinea Pig Serum, Rabbit IgG and Rabbit 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 Guinea Pig anti-Rabbit IgG (H+L) Secondary Antibody (Pre-adsorbed)

Western Blot: Guinea Pig anti-Rabbit IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72763] - Western Blot of Guinea Porcine anti-Rabbit IgG (H+L) Secondary antibody (Pre-adsorbed). Lane M: 3 ul Molecular Ladder. Lane 1: Rabbit IgG whole molecule. Lane 2: Rabbit IgG F(ab) Fragment. Lane 3: Rabbit IgG F(c) Fragment. All samples were reduced. Load: 50 ng per lane. Block for 30 min at RT. Primary Antibody: Guinea Porcine anti-Rabbit IgG (H+L) Secondary antibody (Pre-adsorbed) 1:500 for 60 min at RT. Secondary antibody: Anti-Guinea Porcine IgG (GOAT) Peroxidase Conjugated Antibody 1:40,000 in blocking buffer for 30 min at RT. Predicted/Observed Size: 25 and 50 kDa for Rabbit IgG and Serum, 25 kDa for F(c) and F(ab). Rabbit F(c) migrates slightly higher.
ELISA: Guinea Pig anti-Rabbit IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72763] - ELISA Results of Purified Guinea Porcine Anti-Rabbit IgG mx3 Antibody tested against purified Rabbit IgG (Mx Human IgG, Goat IgG, Mouse IgG). Each well was coated in duplicate with 10 ug of Rabbit IgG [Red Line], Human IgG [Green Line], Goat IgG [Blue Line], Mouse IgG [Purple Line]. The working dilution is 1:17,000. 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 3% Fish Gel/PBS Blocking buffer, Goat Anti-Guinea Porcine mx10 HRP conjugated and TMB substrate.
Guinea Pig anti-Rabbit IgG (H+L) Secondary Antibody (Pre-adsorbed)

Guinea Pig anti-Rabbit IgG (H+L) Secondary Antibody (Pre-adsorbed)

Western Blot of Guinea Pig anti-Rabbit IgG (H+L) Secondary Antibody (Pre-adsorbed)

Applications for Guinea Pig anti-Rabbit IgG (H+L) Secondary Antibody (Pre-adsorbed)

Application
Recommended Usage

ELISA

1:20000 - 1:100000

Immunohistochemistry

1:1000 - 1:5000

Western Blot

1:2000 - 1:10000
Application Notes
This secondary antibody is suitable for use in ELISA, immunohistochemistry, and western blot. Specific conditions for reactivity should be optimized by the end user.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Multi-step

Formulation

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

Format

Pre-adsorbed

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 Guinea Pig anti-Rabbit IgG (H+L) Secondary Antibody (Pre-adsorbed)

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 Guinea Pig anti-Rabbit IgG (H+L) Secondary Antibody (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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