Goat anti-Human IgG (H+L) Secondary Antibody [Alkaline Phosphatase] Summary
Description |
Purity > 95% based on SDS-PAGE. Goat serum was obtained from healthy animals of US origin and under the care of a registered veterinarian.
Store undiluted liquid at 2-8C. For storage at -20C, dilute with an equal volume of glycerol to prevent loss of enzymatic activity. Prepare working dilutions prior to use and then discard. |
Immunogen |
This Goat anti-Human IgG (H+L) Secondary Antibody [Alkaline Phosphatase] was developed against purified human IgG (H&L). |
Specificity |
Based on IEP, this Goat anti-Human IgG (H+L) Secondary Antibody [Alkaline Phosphatase] reacts with heavy gamma chains on human IgG and light chains on all human immunoglobulins. |
Isotype |
IgG |
Clonality |
Polyclonal |
Host |
Goat |
Purity |
Affinity purified |
Applications/Dilutions
Dilutions |
- ELISA 1:200-1:5000
- Immunohistochemistry 1:500-1:5000
- Immunohistochemistry-Paraffin
- Western Blot 1:200-1:5000
|
Application Notes |
This conjugate is suitable for all immunoassay applications. |
Reactivity Notes
Based on IEP, no reactivity is observed to non-immunoglobulin human serum immunoglobulins
Packaging, Storage & Formulations
Storage |
Store at 4C. After dilution, add an equal volume of glycerol to prevent loss of enzymatic activity and store at -20C. Avoid repeated freeze-thaw cycles. |
Buffer |
30 mM Triethanolamine, pH 7.2, 5 mM Magnesium Chloride, 0.1 mM Zinc Chloride, 1 % (w/v) BSA, Protease/IgG free |
Preservative |
0.05% Sodium Azide |
Concentration |
1.5 mg/ml |
Purity |
Affinity purified |
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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