Key Product Details

Species Reactivity

Mouse

Applications

Western Blot, Fluorophore-linked immunosorbent assay, Immunocytochemistry/ Immunofluorescence

Label

DyLight 680 (Excitation = 692 nm, Emission = 712 nm)

Antibody Source

Polyclonal Donkey IgG
View all IgG (H+L) Secondary Antibodies »

Product Specifications

Immunogen

Mouse IgG whole molecule

Specificity

This antibody will react with heavy chains of Mouse IgG and with light chains of most Mouse immunoglobulins.

Clonality

Polyclonal

Host

Donkey

Isotype

IgG

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 Mouse 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, Mouse IgG and Mouse Serum

Scientific Data Images for Donkey anti-Mouse IgG (H+L) Secondary Antibody [DyLight 680]

Western Blot: Donkey anti-Mouse IgG (H+L) Secondary Antibody [DyLight 680] [NBP1-72936] - DyLight (R) dyes can be used for two-color western blot detection with low background and high signal.
Donkey anti-Mouse IgG (H+L) Secondary Antibody [DyLight 680]

Donkey anti-Mouse IgG (H+L) Secondary Antibody [DyLight 680]

DyLight(TM) dyes can be used for two-color western blot detection with low background and high signal. Anti-tubulin was detected using a DyLight(TM) 680 conjugate. Anti-TNFa was detected using a DyLight(TM) 800 conjugate. The image was captured using the Odyssey(R) Infrared Imaging System developed by LI-COR.

Applications for Donkey anti-Mouse IgG (H+L) Secondary Antibody [DyLight 680]

Application
Recommended Usage

Fluorophore-linked immunosorbent assay

1:20000

Immunocytochemistry/ Immunofluorescence

1:5000

Western Blot

1:10000
Application Notes
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.

Reviewed Applications

Read 2 reviews rated 5 using NBP1-72936 in the following applications:

Formulation, Preparation, and Storage

Purification

Multi-step

Reconstitution

Reconstitute with 100 ul deionized water (or equivalent).

Formulation

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 mg/ml

Shipping

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

Stability & Storage

Store lyophilized antibody at 4C in the dark. Aliquot reconstituted liquid and store at -20C. Avoid freeze-thaw cycles.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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 [DyLight 680]

Certificate of Analysis

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

Product Specific Notices for Donkey anti-Mouse IgG (H+L) Secondary Antibody [DyLight 680]



DyLight (R) is a trademark of Thermo Fisher Scientific Inc. and its subsidiaries.

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.

Citations for Donkey anti-Mouse IgG (H+L) Secondary Antibody [DyLight 680]

Customer Reviews for Donkey anti-Mouse IgG (H+L) Secondary Antibody [DyLight 680] (2)

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  • Donkey anti-Mouse IgG (H+L) Secondary Antibody [DyLight 680]
    Name: Anonymous
    Application: Western Blot
    Sample Tested: B cell lymphoma
    Species: Mouse
    Verified Customer | Posted 08/29/2017
    Donkey anti-Mouse IgG (H+L) Secondary Antibody [DyLight 680] NBP1-72936
  • Donkey anti-Mouse IgG (H+L) Secondary Antibody [DyLight 680]
    Name: Christopher England
    Verified Customer | Posted 12/05/2016
    CD20 expression was examined in four lymphoma cell lines. The primary antibody was rabbit and the B-actin antibody was mouse. Dylight 680 donkey anti-mouse IgG was used as the fluorescent secondary to stain B-actin.
    Donkey anti-Mouse IgG (H+L) Secondary Antibody [DyLight 680] NBP1-72936

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Protocols

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FAQs

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