Rabbit F(ab')2 anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed)

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-62587

Novus Biologicals, part of Bio-Techne

Key Product Details

Species Reactivity

Mouse

Applications

Dot Blot, ELISA, Immunohistochemistry, SDS-Page, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit 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 Specifications

Immunogen

Rabbit F(ab')2 anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) was produced by repeated immunization with Mouse IgG whole molecule in goat.

Specificity

No reaction was observed against anti-Pepsin, anti-Rabbit IgG F(c) or Human Serum Proteins.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Description

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, pepsin digestion and chromatographic separation. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Rabbit Serum, Mouse IgG and Mouse 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 F(ab')2 anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed)

Dot Blot: Rabbit F(ab')2 anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP2-62587] - Dot Blot: Rabbit F(ab')2 anti-Mouse IgG (H+L) Secondary Antibody [FITC] (Pre-absorbed) [NBP2-60695] - Dot Blot results of Rabbit F(ab')2 anti-Mouse IgG (H+L) Secondary Antibody [PE] (Pre-absorbed). Dots are Mouse IgG at (1) 100 ng, (2) 33.3 ng, (3) 11.1 ng, (4) 3.70 ng, (5) 1.23 ng. Incubated for 1hr at RT; blocked for 60 min at RT. Imaged with BioRad ChemiDoc, Rhodamine filter. Image from the PE version of this antibody. Image using the FITC form of this antibody.
SDS-PAGE: Rabbit F(ab')2 anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP2-62587]

SDS-PAGE: Rabbit F(ab')2 anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP2-62587]

SDS-Page: Rabbit F(ab')2 anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP2-62587] - SDS-PAGE of Rabbit F(ab')2 anti-Mouse IgG (H+L) Secondary Antibody [Unconjugated] (Pre-adsorbed). Lane 1: Reduced Rabbit F(ab')2 anti-Mouse IgG (H+L) Secondary Antibody [Unconjugated] (Pre-adsorbed). Lane 2: 5uL protein molecular weight standard. Lane 3: Non-reduced Rabbit F(ab')2 anti-Mouse IgG (H+L) Secondary Antibody [Unconjugated] (Pre-adsorbed). Load: 1ug per lane. Predicted/Observed size: Non-reduced at 75 kDa , Reduced at 25 kDa.

Applications for Rabbit F(ab')2 anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed)

Application
Recommended Usage

ELISA

1:2000 - 1:20000

Immunohistochemistry

1:1000 - 1:5000

Western Blot

1:1000 - 1:10000
Application Notes
This product has been tested by ELISA and SDS-PAGE and is suitable for immunomicroscopy and flow cytometry or FACS analysis as well as other antibody based fluorescent assays requiring extremely low background levels, absence of F(c) mediated binding, lot-to-lot consistency, high titer and specificity. The maximum amount of reagent required to stain 1 x 10E6 cells in flow cytometry is approximately 1.0 ug of antibody. Lesser amounts of reagent may be sufficient for staining. Optimal titers for other applications should be determined by the researcher. As a general guideline dilutions of 1:100 to 1:250 should be suitable for most applications.

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

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Certificate of Analysis

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Product Specific Notices for Rabbit F(ab')2 anti-Mouse 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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