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Goat anti-Mouse IgG (H+L) Secondary Antibody [PE] (Pre-adsorbed)

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Dot Blot: Goat anti-Mouse IgG (H+L) Secondary Antibody [PE] (Pre-adsorbed) [NB7594] - Dot Blot of Goat anti-Mouse IgG (H+L) Secondary Antibody [PE] (Pre-adsorbed). Antigen: Mouse IgG. Load: 3-fold serial dilution ...read more
Dot Blot of Goat anti-Mouse IgG Antibody Phycoerythrin Conjugated. Antigen: Mouse IgG. Load: 3-fold serial dilution starting at 200 ng. Primary antibody: None. Secondary antibody: Phycoerythrin goat secondary ...read more

Product Details

Summary
Reactivity MuSpecies Glossary
Applications DB, Flow, ICC/IF
Clonality
Polyclonal
Host
Goat
Conjugate
PE
Format
Pre-adsorbed

Order Details

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Goat anti-Mouse IgG (H+L) Secondary Antibody [PE] (Pre-adsorbed) Summary

Description
This product is stable at 4C as an undiluted liquid. Dilute only prior to immediate use. Centrifuge product if not completely clear after standing at room temperature. Do not freeze after reconstitution. Store reagent in the dark. Use subdued lighting during handling and incubation of cells prior to analysis.

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, conjugation to phycoerythrin and subsequent purification, and extensive dialysis against the buffer stated above. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Phycoerythrin, anti-Goat Serum, Mouse IgG and Mouse Serum
Immunogen
Goat anti-Mouse IgG (H+L) Secondary Antibody [PE] (Pre-adsorbed) was produced by repeated immunization with Mouse IgG whole molecule in goat.
Specificity
No reaction was observed against Bovine, Chicken, Goat, Guinea Pig, Hamster, Horse, Human, Rabbit, Rat and Sheep Serum Proteins.
Isotype
IgG
Clonality
Polyclonal
Host
Goat
Purity
Multi-step

Applications/Dilutions

Dilutions
  • Dot Blot
  • Flow Cytometry 1:100-1:250
  • Immunocytochemistry/ Immunofluorescence 1:100-1:250
Application Notes
This product has been tested by western blot and dot blot and is suitable for immunomicroscopy and flow cytometry or FACS analysis as well as other antibody based fluorescent assays requiring extremely low background levels, absence of F(c) mediated binding, lot-to-lot consistency, high titer and specificity. The maximum amount of reagent required to stain 1 x 10E6 cells in flow cytometry is approximately 1.0 ug of antibody conjugate. Lesser amounts of reagent may be sufficient for staining. Optimal titers for other applications should be determined by the researcher. As a general guideline dilutions of 1:100 to 1:250 should be suitable for most applications.

Packaging, Storage & Formulations

Storage
Store at 4C. Do not freeze.
Buffer
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 10 mg/mL Bovine Serum Albumin (BSA) - Immunoglobulin and Protease free
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.

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Video Protocols

ICC/IF Video Protocol

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