Donkey anti-Goat IgG (H+L) Secondary Antibody [DyLight 649] Summary
| Description |
Store vial at 4C prior to restoration. For extended storage aliquot contents and freeze at -20C or below. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use.
This product was prepared from monospecific antiserum by immunoaffinity chromatography using Goat IgG coupled to agarose beads followed by conjugation to fluorochrome and extensive dialysis against the buffer stated above. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Donkey Serum, Goat IgG and Goat Serum |
| Immunogen |
Goat IgG, whole molecule |
| Specificity |
This antibody will react with heavy chains of Goat IgG and with light chains of most Goat immunoglobulins. |
| Isotype |
IgG |
| Clonality |
Polyclonal |
| Host |
Donkey |
| Purity |
Multi-step |
Applications/Dilutions
| Dilutions |
- Fluorophore-linked immunosorbent assay 1:20000
- Immunocytochemistry/ Immunofluorescence 1:5000
- Western Blot 1:10000
|
| Application Notes |
This product has been tested by dot blot. This product is designed for immunofluorescence microscopy, fluorescence based plate assays (FLISA) and fluorescent western blotting. This product is also suitable for multiplex analysis, including multicolor imaging, utilizing various commercial platforms. The emission spectra for this DyLight(TM) conjugate match the principle output wavelengths of most common fluorescence instrumentation. |
Packaging, Storage & Formulations
| Storage |
Store lyophilized antibody at 4C in the dark. Aliquot reconstituted liquid and store at -20C. Avoid freeze-thaw cycles. |
| Buffer |
Lyophilized from 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 |
| Concentration |
LYOPH |
| Purity |
Multi-step |
| Reconstitution Instructions |
Reconstitute with 100 ul deionized water (or equivalent). |
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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Blogs on IgG (H+L)
![Western Blot: Donkey anti-Goat IgG (H+L) Secondary Antibody [DyLight 649] [NBP1-72846] - color western blot detection with low background and high signal. Anti-tubulin was detected using a DyLight 549 conjugate. Anti-TNFa was detected using a DyLight 649 conjugate. The image was captured using the Imaging System.](http://images.novusbio.com/fullsize/Donkey-anti-Goat-IgG-H+L-Secondary-Antibody-DyLight-649-Western-Blot-NBP1-72846-img0001.jpg "Western Blot: Donkey anti-Goat IgG (H+L) Secondary Antibody [DyLight 649] [NBP1-72846] - color western blot detection with low background and high signal. Anti-tubulin was detected using a DyLight 549 conjugate. Anti-TNFa was detected using a DyLight 649 conjugate. The image was captured using the Imaging System.")
