Sheep anti-Rat IgG (H+L) Secondary Antibody [HRP]

Sheep anti-Rat IgG (H+L) Secondary Antibody [HRP] [NBP2-22109] - Western Blot analysis using Sheep anti-Rat IgG (H+L) Secondary antibody [HRP] (50ng/lane). Lane M: Molecular Ladder Lane 1: Rat IgG whole molecule Lane 2: Rat IgG F(c) fragment Lane 3: Rat IgG Fab fragment Lane 4: Rat IgM whole molecule Lane 5: Rat Serum Primary antibody: at 1:1000 for 60min at RT. Secondary antibody: Donkey anti-Sheep IgG (Peroxidase) at 1:40,000 in blocking buffer for 30min at RT. Block: Blocking buffer for 30min at RT

• Reactivity: Rt
• Applications: WB, ELISA, IHC
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: HRP
• Concentration: 2 mg/ml

Sheep anti-Rat IgG (H+L) Secondary Antibody [HRP] Summary

• Description: To avoid repeated freezing and thawing, aliquot reconstituted stock solution and store at -20C. Dilute required amount only prior to immediate use. Further dilutions can be made in assay buffer. Note: Sodium azide is a potent inhibitor of peroxidase and should not be added to HRP conjugates. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. and prior to removing the cap.
• Immunogen: Rat IgG whole molecule
• Specificity: Recognizes rat IgG. Assay by Immunoelectrophoresis resulted in a single precipitin arc against anti-peroxidase, anti-sheep serum, rat IgG and rat serum.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • ELISA 1:5000-1:100000
  • Immunohistochemistry 1:500-1:2500
  • Western Blot 1:1000-1:10000
• Application Notes: ELISA: Assayed against 1ug of Rat IgG in a standard capture ELISA using ABTS as a substrate for 30 minutes at RT. Optimal working dilutions to be determined by researcher.
• Readout System:
  • NovaLume Plus Chemiluminescent Reagent LUM-555

Packaging, Storage & Formulations

• Storage: Store at 4C in the dark.
• Buffer: PBS, pH 7.2 with 10mg/ml BSA (IgG and protease free)
• Preservative: 0.01% Gentamicin Sulfate
• Concentration: 2 mg/ml
• Purity: Immunogen affinity purified

Notes

To avoid repeated freezing and thawing, aliquot reconstituted stock solution and store at -20C. Dilute required amount only prior to immediate use. Further dilutions can be made in assay buffer. Note: Sodium azide is a potent inhibitor of peroxidase and should not be added to HRP conjugates. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. and prior to removing the cap.

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.

FAQs for IgG (H+L) Antibody (NBP2-22109). (Showing 1 - 1 of 1 FAQ).

1. We have a question about the sheep anti-Rat IgE Antibody (NBP2-22109).The datasheet of the product only label the content of antibody as 0.5mg, but the antibody is in liquid. So we didn't know the concentration or the volume of the antibody. Can NOVUS could provide this information? Please reply me as soon as possible, thank you very much?
   • This antibody has a concentration of 2.0 mg/ml. With 0.5mg, the volume should be 0.25ml.