Rabbit anti-Porcine IgG (H+L) Secondary Antibody

Western Blot: Rabbit anti-Porcine IgG (H+L) Secondary Antibody [NBP1-73812] - Lane 1: Swine IgG Load: 100 ng per lane Primary antibody: Porcine IgG (H+L) Antibody incubated overnight at 4C. Secondary antibody: Rabbit anti-Porcine IgG (H+L) Secondary Antibody incubated for 30 min at RT; incubated with blocking buffer for 30 min at RT. Predicted/Observed size: 55 and 28 kDa, 55 and 28 kDa.

• Reactivity: Po
• Applications: WB, ELISA, IHC
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: LYOPH

Rabbit anti-Porcine IgG (H+L) Secondary Antibody Summary

• Description: This product was prepared from monospecific antiserum by a delipidation and defibrination. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-rabbit serum, Swine IgG and Swine Serum.
  
  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.
• Immunogen: Swine IgG whole molecule
• Isotype: Serum
• Clonality: Polyclonal
• Host: Rabbit
• Purity: Delipidation and Defibrination

Applications/Dilutions

• Dilutions:
  • ELISA 1:20000 - 1:100000
  • Immunohistochemistry 1:1000 - 1:5000
  • Western Blot 1:2000 - 1:10000
• Application Notes: This product has been tested in western blot and is suitable for use in ELISA and immunohistochemistry. Specific conditions for reactivity should be optimized by the end user.

Packaging, Storage & Formulations

• Storage: Store lyophilized antibody at 4C. Aliquot reconstituted liquid and store at -20C. Avoid freeze-thaw cycles.
• Buffer: Lyophilized from 0.01 M Sodium Phosphate, 0.15 M Sodium Chloride, pH 7.2
• Preservative: 0.01% Sodium Azide
• Concentration: LYOPH
• Purity: Delipidation and Defibrination
• Reconstitution Instructions: Reconstitute with 2.0 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.

Publications for IgG (H+L) Antibody (NBP1-73812)(2)

Publications using NBP1-73812

• Wiarda J, Shircliff A, Stasko J et al. Conserved B cell signaling, activation, and differentiation in porcine jejunal and ileal Peyer's patches despite distinct immune landscapes bioRxiv 2023-09-15 (IHC)
• Wiarda JE, Shircliff AL, Becker SR et Al. Conserved B cell signaling, activation, and differentiation in porcine jejunal and ileal Peyer's patches despite distinct immune landscapes Mucosal Immunol 2024-08-13 [PMID: 39147277]