Goat anti-Armenian Hamster IgG (H+L) Secondary Antibody

Western Blot: Goat anti-Armenian Hamster IgG (H+L) Secondary Antibody [NB120-5738] - Western Blot: Goat anti-Armenian Hamster IgG Secondary Antibody [FITC] [NB120-5743] - Lane 1: Armenian Hamster IgG. Lane 2: none. Load: 50 ng per lane. Image using the FITC form of this antibody.

Western Blot: Goat anti-Armenian Hamster IgG (H+L) Secondary Antibody [NB120-5738] - Lane 1: Armenian Hamster IgG. Lane 2: none. Load: 50 ug per lane. Primary antibody: Armenian Hamster IgG Biotin conjugated antibody at 1:1,000 for overnight at 4C. Secondary antibody: HRP Streptavidin secondary antibody at 1:40,000 for 30 min at RT. Blocking buffer for 30 min at RT. Predicted/Observed size: 55 kDa, 28 kDa for Human IgG.

Western Blot: Goat anti-Armenian Hamster IgG (H+L) Secondary Antibody [NB120-5738] - Western Blot: Goat anti-Armenian Hamster IgG (H+L) Secondary Antibody [Texas Red] [NBP1-73458] - Lane 1: Armenian Hamster IgG. Lane 2: None. Load: 50 ng per lane. Primary antibody: None. Secondary antibody: Texas Red goat secondary antibody at 1:1,000 for 60 min at RT. Block: incubated with blocking buffer for 30 min t RT. Predicted/Observed size: 25 & 55 kDa, 25 & 55 kDa for Armenian Hamster IgG. Other band(s): None.

ELISA: Goat anti-Armenian Hamster IgG (H+L) Secondary Antibody [NB120-5738] - ELISA Results of Goat anti-Armenian Hamster IgG (H+L) Secondary antibody tested against purified Armenian Hamster IgG. Each well was coated in duplicate with 1.0 ug of Armenian Hamster IgG. The working dilution is 1:21,000. The starting dilution of antibody was 5ug/ml and the X-axis represents the Log10 of a 3-fold dilution. This titration is a 4-parameter curve fit where the IC50 is defined as the titer of the antibody. Assay performed using HRP Conjugate Substrate, Donkey Anti-Goat IgG HRP conjugated, and TMB substrate.

• Reactivity: ArHa
• Applications: WB, DB, ELISA, IHC, IP, ChIP
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated

Goat anti-Armenian Hamster IgG (H+L) Secondary Antibody Summary

• Description: This product was prepared from monospecific antiserum by immunoaffinity chromatography using Armenian Hamster 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-Goat Serum, Armenian Hamster IgG and Armenian Hamster Serum
  
  Store vial at 4C prior to opening. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use. For extended storage aliquot contents and freeze at -20C or below. Avoid cycles of freezing and thawing.
• Immunogen: Armenian Hamster IgG whole molecule
• Specificity: Reactivity with Golden Syrian Hamster IgG is greatly reduced.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Purity: Multi-step

Applications/Dilutions

• Dilutions:
  • Chromatin Immunoprecipitation (ChIP)
  • Dot Blot
  • ELISA 1:20000 - 1:100000
  • Immunohistochemistry 1:1000 - 1:5000
  • Immunoprecipitation
  • Western Blot 1:2000 - 1:10000
• Application Notes: This product has been tested by ELISA and is suitable for immunoprecipitation, immunodiffusion, conjugation and most immunological methods requiring lot-to-lot consistency, high titer and specificity.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
• Preservative: 0.01% Sodium Azide
• Purity: Multi-step

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 (NB120-5738)(2)

Publications using NB120-5738

• Iijima N, Yamaguchi M, Hayashi T et al. miR-147-3p in pathogenic CD4 T cells controls chemokine receptor expression for the development of experimental autoimmune diseases. Journal of autoimmunity 2024-10-11 [PMID: 39395343]
• Zhu X, Trimarco JD, Williams CA Et al. ZBTB7A promotes virus-host homeostasis during human coronavirus 229E infection Cell Rep 2022-10-15 [PMID: 36243002] (WB)
  
  Details:
  Citation using the HRP version of this antibody.