Key Product Details

Species Reactivity

Mouse

Applications

Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Western Blot

Label

FITC (Excitation = 495 nm, Emission = 519 nm)

Antibody Source

Polyclonal Donkey IgG

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

View all IgG (H+L) Secondary Antibodies »

Product Specifications

Immunogen

Mouse IgG whole molecule

Clonality

Polyclonal

Host

Donkey

Isotype

IgG

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-Fluorescein, anti-Donkey Serum, Mouse IgG and Mouse Serum.

Scientific Data Images for Donkey anti-Mouse IgG (H+L) Secondary Antibody [FITC]

Western Blot: Donkey anti-Mouse IgG (H+L) Secondary Antibody [FITC] [NB120-6816] - Western Blot of Fluorescein Donkey anti-Mouse IgG (H+L) Secondary Antibody [FITC]. Lane 1: Mouse IgG.Lane 2: None.Load: 50 ng per lane.Primary antibody: None.Secondary antibody: Donkey anti-Mouse IgG (H+L) Secondary Antibody [FITC] at 1:1000 for 60 min at RT.Blocking: Fluorescent blocking buffer for 30 min at RT.Predicted/Observed size: 25 & 55 kDa, 25 & 55 kDa for Mouse IgG.Other band(s): None.
Donkey anti-Mouse IgG (H+L) Secondary Antibody [FITC]

Donkey anti-Mouse IgG (H+L) Secondary Antibody [FITC]

Western Blot of Fluorescein Donkey Anti-Mouse IgG secondary antibody. Lane 1: Mouse IgG. Lane 2: None. Load: 50 ng per lane. Primary antibody: None. Secondary antibody: Fluorescein donkey secondary antibody at 1:1,000 for 60 min at RT. Blocking: for 30 min at RT. Predicted/Observed size: 25 & 55 kDa, 25 & 55 kDa for Mouse IgG. Other band(s): None.

Applications for Donkey anti-Mouse IgG (H+L) Secondary Antibody [FITC]

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:500 - 1:2500

Western Blot

1:1000 - 1:10000
Application Notes
This product has been tested by dot blot and 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.

Formulation, Preparation, and Storage

Purification

Multi-step

Formulation

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

Please see the vial label for concentration. If unlisted please contact technical services.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: IgG (H+L)

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

Additional IgG (H+L) Products

Product Documents for Donkey anti-Mouse IgG (H+L) Secondary Antibody [FITC]

Product Datasheets

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

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Product Specific Notices for Donkey anti-Mouse IgG (H+L) Secondary Antibody [FITC]

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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