Key Product Details

Species Reactivity

Sheep

Applications

Dot Blot, ELISA, Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot

Label

Biotin

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

Sheep IgG whole molecule

Specificity

This antibody was pre-adsorbed against Human Serum Proteins. No reaction was observed against Human Serum Proteins.

Clonality

Polyclonal

Host

Rabbit

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 Sheep 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-biotin, anti-Rabbit Serum, Sheep IgG and Sheep Serum

Scientific Data Images

Dot Blot: Rabbit anti-Sheep IgG (H+L) Secondary Antibody [Biotin] (Pre-adsorbed) [NB7584] - Dot Bot of Rabbit anti-Sheep IgG (H+L) Secondary antibody [Biotin] (Pre-adsorbed).Lane 1: 100ng.Lane 2: 33.3ng.Lane 3: 11.1ng.Lane 4: 3.7ng.Lane 5: 1.23ng.Primary Antibody: Rb-anti-sheep Biotin at 1ug/mL.Secondary Antibody: Streptavidin HRP at 1:40,000.Blocking Buffer for 30 min at RT.
ELISA: Rabbit anti-Sheep IgG (H+L) Secondary Antibody [Biotin] (Pre-adsorbed) [NB7584] - ELISA results of purified Rabbit anti-Sheep IgG (H+L) Secondary antibody [Biotin] (Pre-adsorbed) antibody Min X human serum tested against purified Sheep IgG. Each well was coated in duplicate with 1.0 ug of sheep 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 Blocking buffer, Streptavidin HRP conjugate 1:10, 000, and TMB substrate.
Rabbit anti-Sheep IgG (H+L) Secondary Antibody [Biotin] (Pre-adsorbed)

Rabbit anti-Sheep IgG (H+L) Secondary Antibody [Biotin] (Pre-adsorbed)

Dot Bot of Rabbit anti-Sheep IgG (H+L) Secondary Antibody [Biotin] (Pre-adsorbed). Lane 1: 100ng. Lane 2: 33.3ng. Lane 3: 11.1ng. Lane 4: 3.7ng. Lane 5: 1.23ng. Primary Antibody: Rb-anti-sheep Biotin at 1ug/mL. Secondary Antibody: Streptavidin HRP S000-03 at 1:40,000. Blocking Buffer: BlockOut for 30 min at RT.

Applications

Application
Recommended Usage

ELISA

1:20000 - 1:100000

Immunohistochemistry

1:1000 - 1:5000

Immunohistochemistry-Paraffin

1:500-1:1000

Western Blot

1:2000 - 1:10000
Application Notes
This product has been tested by ELISA and Dot Blot and is suitable for western blot, ELISA and immunohistochemistry as well as other antibody based assays requiring lot-to-lot consistency.

Formulation, Preparation, and Storage

Purification

Multi-step

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

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

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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