Donkey anti-Rat IgG (H+L) Secondary Antibody [HRP] (Pre-adsorbed)

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Western Blot: Donkey anti-Rat IgG (H+L) Secondary Antibody [HRP] (Pre-adsorbed) [NBP1-73749] - Donkey anti-Rat IgG (H+L) Secondary Antibody [HRP] [NBP1-73749] - Lane 1: Rat IgG whole molecule. Lane 2: Rat IgG F(c) ...read more

Product Details

Summary
Reactivity RtSpecies Glossary
Applications WB, ELISA, IHC, IHC-Fr, IHC-P
Clonality
Polyclonal
Host
Donkey
Conjugate
HRP
Format
Pre-adsorbed
Concentration
LYOPH

Order Details

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

Immunogen
This Donkey anti-Rat IgG (H+L) Secondary Antibody [HRP] (Pre-adsorbed) was developed against rat IgG whole molecule.
Specificity
Assay of Donkey anti-Rat IgG (H+L) Secondary Antibody [HRP] (Pre-adsorbed) by immunoelectrophoresis resulted in a single precipitin arc against anti-Peroxidase, anti-Donkey serum, Rat IgG and Rat serum. No reaction was observed against Bovine, Chicken, Goat, Guinea Pig, Hamster, Horse, Human, Mouse, Rabbit and Sheep serum proteins. This antibody was pre-adsorbed against bovine chicken goat guinea pig hamster horse human mouse rabbit & sheep serum proteins.
Isotype
IgG
Clonality
Polyclonal
Host
Donkey
Purity
Immunogen affinity purified

Applications/Dilutions

Dilutions
  • Western Blot 1:1000 - 1:10000
  • ELISA 1:10000 - 1:50000
  • Immunohistochemistry 1:500 - 1:2500
  • Immunohistochemistry-Frozen 1:500 - 1:2500
  • Immunohistochemistry-Paraffin 1:500 - 1:2500
Application Notes
This product has been assayed against 1.0 ug of Rat IgG in a standard capture ELISA using ABTS ELISA PEROXIDASE SUBSTRATE for 30 minutes at room temperature. A working dilution of 1:15000 to 1:60000 is suggested for this product.

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

Packaging, Storage & Formulations

Storage
Store at 4C in the dark.
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
Immunogen affinity purified
Reconstitution Instructions
Reconstitute with 1.0 ml deionized water

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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Product General Protocols

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WB Video Protocol

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Other Available Formats

Alkaline Phosphatase NBP1-72681
Allophycocyanin/Cy7 NBP1-72743APCCY7
Biotin NBP1-73684
DyLight 405 NBP1-72743V
DyLight 488 NBP1-72743G
DyLight 550 NBP1-72743R
DyLight 594 NBP1-72743DL594
DyLight 650 NBP1-72743C
DyLight 680 NBP1-72743FR
DyLight 755 NBP1-72743IR
FITC NBP1-73683
HRP NBP1-73749
Janelia Fluor 549 NBP1-72743JF549
Janelia Fluor 646 NBP1-72743JF646
PE/Atto594 NBP1-72743PEATT594
PE/Cy5.5 NBP1-72743PECY55
PE/Cy7 NBP1-72743PECY7
Rhodamine NBP1-73569
Texas Red NBP1-73685

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