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

Western Blot: Goat anti-Hamster IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-71730] - Golden Syrian Hamster IgG (Lane 2) and Armenian Hamster IgG (Lane 3) was run on a 4-20% gel and transferred to 0.45 um nitrocellulose. After blocking with 5% Blotto 30 min at 20C, Antibody was used at 1:5000 in blocking buffer for Fluorescent Western blotting. Molecular weight markers are in lane 1.

Immunocytochemistry/Immunofluorescence: Goat anti-Hamster IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-71730] - This image shows anti-histone detection using a DyLight 488 conjugate (green). Anti-Tubulin was detected using a DyLight 549 conjugate (red). Nuclei were counter-stained using DAPI (blue). The image was captured using an Axio Imager.Z1 (Zeiss Micro Imaging Inc).. Using the DyLight 488 format of this antibody.

Western Blot: Goat anti-Hamster IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-71730] - Anti-TNF-alpha was detected using a DyLight 800 conjugate. The image was captured using the Odyssey Infrared Imaging System developed by LI-COR.

ELISA: Goat anti-Hamster IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-71730] - ELISA results of purified Goat anti-Hamster IgG (H+L) Secondary Antibody [Unconjugated] (Pre-adsorbed) tested against purified Golden Syrian Hamster IgG and Armenian Hamster IgG. Each well was coated in duplicate with 1.0 ug of protein from different species. Golden Syrian Hamster IgG (Green Line), Armenian Hamster IgG (Red Line), Mouse IgG (Blue Line), and Rat IgG (Purple Line). 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 Blocking buffer

Dot Blot: Goat anti-Hamster IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-71730] - Dot Blot for Goat anti-Hamster IgG (H+L) Secondary Antibody [Texas Red] (Pre-adsorbed). Lane 1: 100 ng Hamster IgG. Lanes 2-5: serial dilution 3 fold of Antigen. Primary Antibody: none.Secondary Antibody: Goat anti-Hamster IgG (H+L) Secondary Antibody [Texas Red] (Pre-adsorbed) at 1:1000 dilution at RT for 1 hour. Block: Fluorescent blocking buffer at RT for 30 minutes.. Using the Texas Red format of this antibody.

Fluorophore-linked immunosorbent assay: Goat anti-Hamster IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-71730] - Using the DyLight 405 format of this antibody.

• Reactivity: Ha
• Applications: WB, DB, ELISA, ICC/IF, IHC
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Format: Pre-adsorbed

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

• Description: This product was prepared from monospecific antiserum by immunoaffinity chromatography using Golden Syrian and 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, Golden Syrian Hamster IgG and Armenian Hamster IgG
  
  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 and Golden Syrian Hamster IgG, whole molecule
• Specificity: This antibody was pre-adsorbed against Mouse and Rat Serum Proteins.
  
  No reaction was observed against Mouse or Rat serum proteins. Specificity was confirmed using ELISA at less than 1% cross reactivity.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Purity: Multi-step

Applications/Dilutions

• Dilutions:
  • Dot Blot
  • ELISA 1:20000 - 1:100000
  • Fluorophore-linked immunosorbent assay
  • Immunocytochemistry/ Immunofluorescence
  • Immunohistochemistry 1:1000 - 1:5000
  • Western Blot 1:2000 - 1:10000
• Application Notes: This product has been tested by ELISA and dot blot and is suitable for ELISA, western blot, and immunohistochemistry, as well as other assays requiring lot-to-lot consistency.

Reactivity Notes

Cricetulus migratorius (Armenian Hamster); Melanochromis auratus (Golden Syrian Hamster)

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 (NBP1-71730)(3)

Publications using NBP1-71730

• Trzilova D, Anjuwon-Foster BR, Torres Rivera D, Tamayo R Rho factor mediates flagellum and toxin phase variation and impacts virulence in Clostridioides difficile PLoS Pathog. 2020-08-01 [PMID: 32785266] (Hamster)
• Liang Q, Zhang L, Wood RW Et al. Avian Reticuloendotheliosis Viral Oncogene Related B Regulates Lymphatic Endothelial Cells during Vessel Maturation and Is Required for Lymphatic Vessel Function in Adult Mice Am J Pathol 2019-09-21 [PMID: 31539516]
  
  Details:
  Citation using the DyLight 405 version of this antibody.
• Anjuwon-Foster BR, Maldonado-Vazquez N, Tamayo R. Et al. Characterization of Flagellum and Toxin Phase Variation in Clostridioides difficile Ribotype 012 Isolates J Bacteriol 2018-05-09 [PMID: 29735765] (WB, WB)
  
  Details:
  Citation using the DyLight 800 version of this antibody.