Rabbit anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed)

Western Blot: Rabbit anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72731] - Western blot expression of phospho PDH, PDK1, LDH5 and HIF1 alpha in hepatoma HepG2 cells, at 0, 30 and 60 min following exposure to 100 ug/ml of amifostine. Secondary antibody used: Rabbit anti-Mouse IgG (H+L) Secondary Antibody (NBP1-72731). Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep30986) licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: Rabbit anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72731] - Hep core expression in Huh7 cells. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature. The secondary antibody was diluted in 1% BSA in PBS at 1:500 dilution for 20 minutes at room temperature. Image from verified customer review. Image from the DyLight 488 version of this antibody.

Western Blot: Rabbit anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72731] - Western Blot: Rabbit anti-Mouse IgG Secondary Antibody [Rhodamine] [NB120-6725] - Lane 1: Mouse IgG. Lane 2: None. Load: 50 ng per lane.

Immunocytochemistry/Immunofluorescence: Rabbit anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72731] - Immunocytochemistry/Immunofluorescence: Rabbit anti-Mouse IgG (H+L) Secondary Antibody [DyLight 488] [NBP1-72889] Image from the DyLight 488 version of this antibody.

Fluorophore-linked immunosorbent assay: Rabbit anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72731]

ELISA: Rabbit anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72731] - ELISA results of purified Rabbit anti-Mouse IgG (H+L) Secondary antibody (Pre-adsorbed) tested against purified Mouse IgG. Each well was coated in duplicate with 1.0 ug of Mouse IgG [red line] and Human IgG [blue 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 3% fish gelatin, Goat anti-Rabbit igG Antibody Peroxidase Conjugated (Min X Bv Ch Gt GP Ham Hs Hu Ms Rt & Sh Serum Proteins) and TMB ELISA Substrate.

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

Rabbit anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) Summary

• Description: This product was prepared from monospecific antiserum by immunoaffinity chromatography using Mouse IgG coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities. Assay by immunoelectrophoresis (IEP) resulted in a single precipitin arc against anti-Rabbit Serum, Mouse IgG and Mouse 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: Rabbit anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) was produced by repeated immunization with mouse IgG whole molecule in rabbit
• Specificity: This antibody was pre-adsorbed against Human Serum Proteins.
  
  No reaction was observed against Human Serum Proteins.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Purity: Multi-step

Applications/Dilutions

• Dilutions:
  • ELISA 1:20000 - 1:40000
  • 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 is suitable for immunoblotting (western or dot blot), ELISA, and immunohistochemistry as well as other peroxidase-antibody based enzymatic assays requiring lot-to-lot consistency.

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 (NBP1-72731)(3)

Publications using NBP1-72731

• Shengjie Zhang, Eui-Jun Kim, Junfeng Huang, Peng Liu, Kristine Donahue, Qinchuan Wang, Yidan Wang, Sean Mcilwain, Ling Xie, Xian Chen, Lingjun Li, Wei Xu NEAT1 repression by MED12 creates chemosensitivity in p53 wild-type breast cancer cells. The FEBS journal 2024-02-21 [PMID: 38380720]
• Ducas AA, Kuhn DCS, Bath LC Et al. Increased matrix metalloproteinase 9 activity correlates with flow-mediated intraluminal thrombus deposition and wall degeneration in human abdominal aortic aneurysm JVS Vasc Sci 2021-10-07 [PMID: 34617048]
  
  Details:
  Citation using the Biotin version of this antibody.
• Koukourakis MI, Giatromanolaki A, Zois CE et al. Normal tissue radioprotection by amifostine via Warburg-type effects Sci Rep 2016-08-11 [PMID: 27507219] (WB)