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Mouse anti-Rabbit IgG (H+L) Secondary Antibody [HRP] (Pre-adsorbed)

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Western Blot: Mouse anti-Rabbit IgG (H+L) Secondary Antibody [HRP] (Pre-adsorbed) [NBP1-73746] - Western blot of Mouse anti-Rabbit IgG (H+L) Secondary Antibody [HRP] (Pre-adsorbed).Lane 1: Rabbit IgG.Lane 2: ...read more
Western Blot: Mouse anti-Rabbit IgG (H+L) Secondary Antibody [HRP] (Pre-adsorbed) [NBP1-73746] - Lane 1: Rabbit IgG whole molecule. Lane 2: Rabbit IgG F(ab) Fragment. Lane 3: Rabbit IgG F(c) Fragment. Lane 4: Rabbit IgM ...read more
Western blot of Peroxidase conjugated Mouse Anti-Rabbit IgG Pre-Adsorbed secondary antibody. Lane 1: Rabbit IgG. Lane 2: None. Load: 50 ng per lane. Primary antibody: None. Secondary antibody: Peroxidase mouse secondary ...read more
Western Blot of Mouse anti-Rabbit IgG (H+L) Secondary Antibody [HRP] (Pre-adsorbed)

Product Details

Summary
Reactivity RbSpecies Glossary
Applications WB, ELISA, IHC
Clonality
Polyclonal
Host
Mouse
Conjugate
HRP
Format
Pre-adsorbed
Concentration
LYOPH

Order Details

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Mouse anti-Rabbit IgG (H+L) Secondary Antibody [HRP] (Pre-adsorbed) Summary

Description
Store vial at 4C prior to restoration. 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 temperature. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use.

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-Peroxidase, anti-Mouse Serum, Rabbit IgG and Rabbit Serum
Immunogen
Rabbit IgG whole molecule
Specificity
This antibody was pre-adsorbed against Human, Goat, and Mouse Serum Proteins.

No reaction was observed against or Goat, Human and Mouse Serum Proteins.
Isotype
IgG
Clonality
Polyclonal
Host
Mouse
Purity
Multi-step

Applications/Dilutions

Dilutions
  • ELISA 1:10000 - 1:50000
  • Immunohistochemistry 1:500 - 1:2500
  • Western Blot 1:1000 - 1:10000
Application Notes
This product has been tested by ELISA and western blot and is assayed against 1.0 ug of Rabbit IgG in a standard capture ELISA using ABTS (2,2'-azino-bis-[3-ethylbenthiazoline-6-sulfonic acid]) as a substrate for 30 minutes at room temperature. A working dilution of 1:8,000 to 1:32,000 of the reconstitution concentration is suggested for this product.

NOTE: Do NOT add Sodium Azide (it inhibits HRP irreversibly).
Readout System

Packaging, Storage & Formulations

Storage
Store lyophilized antibody at 4C in the dark. Aliquot reconstituted liquid and store at -20C. Avoid freeze-thaw cycles.
Buffer
Lyophilized from 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 10 mg/mL Bovine Serum Albumin (BSA) - Immunoglobulin and Protease free
Preservative
0.01% Gentamicin Sulfate
Concentration
LYOPH
Purity
Multi-step
Reconstitution Instructions
Reconstitute with 1.0 ml deionized water (or equivalent).

Background

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

Limitations

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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