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A3812 Sigma-Aldrich

Anti-Rabbit IgG (whole molecule)–Alkaline Phosphatase antibody produced in goat

affinity isolated antibody, buffered aqueous glycerol solution

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Properties

Related Categories Alkaline Phosphatase, Alkaline Phosphatase Conjugates, Alphabetical Index, Antibodies, Antibody Conjugates,
conjugate   alkaline phosphatase conjugate
clone   polyclonal
biological source   goat
application(s)   direct ELISA: 1:30,000
  dot blot: 1:30,000
  immunohistochemistry (formalin-fixed, paraffin-embedded sections): 1:50
  western blot: 1:30,000
species reactivity   rabbit
form   buffered aqueous glycerol solution
shipped in   wet ice
storage temp.   2-8°C
antibody form   affinity isolated antibody
antibody product type   secondary antibodies
Quality Level   200

Description

General description

Anti-rabbit IgG (whole molecule) is produced in goat using purified rabbit IgG as the immunogen. Antibody is isolated from anti-rabbit IgG antiserum by immunospecific purification which removes essentially all goat serum proteins, including immunoglobulins that do not specifically bind to rabbit IgG. The antibody preparation is solid phase adsorbed with human serum proteins to ensure minimal cross reactivity in tissue or cell preparations. Anti-Rabbit IgG is conjugated to alkaline phosphatase by protein cross linking with 0.2% glutaraldehyde.
Specificity of the antiserum is determined by immunoelectrophoresis prior to conjugation, versus normal rabbit serum and rabbit IgG. Identity and purity of the antibody is established by immunoelectrophoresis prior to conjugation. Electrophoresis of the product followed by diffusion versus anti-goat IgG and anti-goat whole serum results in single arcs of precipitation.

IgGs are glycoprotein antibodies that modulate several immune responses. Rabbit IgGs against target proteins are often used as primary antibodies in various research applications. Thus, secondary anti-rabbit IgGs conjugated to a detectable substrate are useful tools for the analysis of target proteins.

Immunoglobulin G (IgG) belongs to the immunoglobulin family and is a widely expressed serum antibody. It consists of a gamma (γ) heavy chain in the constant (C) region. The monomeric 150kDa structure of IgG constitutes two identical heavy chains and two identical light chains with molecular weight of 50kDa and 25kDa, respectively. The primary structure of this antibody also contains disulfide bonds involved in linking the two heavy chains, linking the heavy and light chains and resides inside the chains. IgG is further subdivided into four classes namely, IgG1, IgG2, IgG3, and IgG4 with different heavy chains, named γ1, γ2, γ3, and γ4, respectively. Limited digestion using papain cleaves the antibody into three fragments, two of which are identical and contain the antigen-binding activity (Fab fragments). The third fragment does not possess antigen-binding activity and is known as fragment crystallizable (Fc).

Immunogen

Purified rabbit IgG

Application

Citrullination of antithrombin by PADI4 was analyzed by ELISA using alkaline phosphatase conjugated goat anti-rabbit IgG as the secondary diluted at 1:5000 in 0.05M carbonate/bicarbonate buffer (Ph 9.6).

Goat anti-rabbit IgG (whole molecule)-alkaline phosphatase antibody can be used for western blot and ELISA

It can also be used for immunohistochemistry (1:50) and dot bot (1:30,000) assays.

Physical form

Solution in 0.05 M Tris buffer, pH 8.0, containing 1 mM MgCl2, 10 mM glycine, 1% bovine serum albumin, 50% glycerol and 15 mM sodium azide

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Preparation Note

Adsorbed to reduce background staining with human samples.

Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport
WGK Germany 
2
Protocols & Articles

Articles

Antibody Basics

Immunoglobulins (Igs) are produced by B lymphocytes and secreted into plasma. The Ig molecule in monomeric form is a glycoprotein with a molecular weight of approximately 150 kDa that is shaped more ...
Keywords: Affinity chromatography, Centrifugation, Chromatography, Digestions, Direct immunofluorescence, Gene expression, High performance liquid chromatography, Immunofluorescence, Ion Exchange, Microscopy, Precipitation, Purification, Rheumatology, Scanning electron microscopy

How to Choose a Secondary Antibody

The following information is provided to help you decide which secondary antibody may be best for your particular application.
Keywords: Amplification, Enzyme-linked immunosorbent assay, Flow cytometry, Immobilization, Immunocytochemistry, Immunofluorescence, Immunohistochemistry, Western blot

Secondary Antibodies, Conjugates and Kits

Secondary antibodies are polyclonal or monoclonal antibodies that bind to primary antibodies or antibody fragments, such as the Fc or Fab regions. They are typically labeled with probes that make the...
Keywords: Absorption, Adsorption, Amplification, Bacterial conjugations, Cancer, Digestions, Electrophoresis, Enzyme-linked immunosorbent assay, Flow cytometry, Gel electrophoresis, Gene expression, Hormones, Immobilization, Immunocytochemistry, Immunofluorescence, Immunohistochemistry, Immunology, Immunoprecipitation, Infrared spectroscopy, Microbiology, Microscopy, Purification, Vitamins, Western blot

Protocols

Western Blot Protocol | Immunoblotting Protocol

Western Blotting refers to the electrophoretic transfer of proteins from sodium dodecyl sulfate polyacrylamide gels to sheets of PVDF or nitrocellullose membrane, followed by immunodetection of prote...
Keywords: AGE, Buffers, Cell disruption, Detection methods, Detergents, Dialysis, Electroblotting, Electrophoresis, Enzyme activity, Gel electrophoresis, Immunoprecipitation, PAGE, Protein extraction, Purification, Sample preparations, Western blot

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Peer-Reviewed Papers
15

References

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