Goat anti-Mouse IgG (H+L) Secondary Antibody [Rhodamine] (Pre-adsorbed) Summary
| Description |
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
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 resulted in a single precipitin arc against anti-Goat Serum, Mouse IgG and Mouse Serum |
| Immunogen |
Mouse IgG whole molecule |
| Specificity |
This antibody was pre-adsorbed against Human Serum Proteins.
No reaction was observed against Human Serum Proteins. |
| Isotype |
IgG |
| Clonality |
Polyclonal |
| Host |
Goat |
| Purity |
Multi-step |
Applications/Dilutions
| Dilutions |
- Flow Cytometry 1:500 - 1:2500
- Fluorophore-linked immunosorbent assay 1:10000 - 1:50000
- Immunocytochemistry/ Immunofluorescence 1:1000 - 1:5000
- Western Blot 1:1000 - 1:10000
|
| Application Notes |
This product has been tested by western blot and 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. |
Packaging, Storage & Formulations
| Storage |
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles. |
| Buffer |
0.01 M Sodium 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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Blogs on IgG (H+L)
![Western Blot: Goat anti-Mouse IgG (H+L) Secondary Antibody [Rhodamine] (Pre-adsorbed) [NBP1-73133] - Western Blot of Goat anti-Mouse IgG (H+L) Secondary Antibody [Rhodamine] (Pre-adsorbed).Lane 1: Mouse IgG. Lane 2: None. Load: 50 ng per lane.Primary antibody: None.Secondary antibody: Goat anti-Mouse IgG (H+L) Secondary Antibody [Rhodamine] (Pre-adsorbed) at 1:1000 for 60 min at RT.Block: Fluorescent blocking buffer for 30 min at RT.Predicted/Observed size: 28 & 55 kDa, 28 & 55 kDa for Mouse IgG. Other band(s): None.](http://images.novusbio.com/fullsize/Goat-anti-Mouse-IgG-H+L-Secondary-Antibody-Rhodamine-Pre-adsorbed-Western-Blot-NBP1-73133-img0006.jpg "Western Blot: Goat anti-Mouse IgG (H+L) Secondary Antibody [Rhodamine] (Pre-adsorbed) [NBP1-73133] - Western Blot of Goat anti-Mouse IgG (H+L) Secondary Antibody [Rhodamine] (Pre-adsorbed).Lane 1: Mouse IgG. Lane 2: None. Load: 50 ng per lane.Primary antibody: None.Secondary antibody: Goat anti-Mouse IgG (H+L) Secondary Antibody [Rhodamine] (Pre-adsorbed) at 1:1000 for 60 min at RT.Block: Fluorescent blocking buffer for 30 min at RT.Predicted/Observed size: 28 & 55 kDa, 28 & 55 kDa for Mouse IgG. Other band(s): None.")
