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Key Product Details

Species Reactivity

Mouse

Applications

Flow Cytometry, Immunomicroscopy

Label

DyLight 594 (Excitation = 593 nm, Emission = 618 nm)

Antibody Source

Polyclonal Donkey IgG

Format

Pre-adsorbed

Concentration

LYOPH mg/ml

Product Specifications

Immunogen

This Donkey anti-Mouse IgG (H+L) Secondary Antibody [DyLight 594] (Pre-adsorbed) was developed against purified mouse IgG (H&L).

Reactivity Notes

Based on IEP, no reactivity is observed to non-immunoglobulin mouse serum immunoglobulins and IgG from bovine, chicken, goat, guinea pig, hamster, horse, human, rabbit, rat, or sheep

Specificity

Based on IEP, this Donkey anti-Mouse IgG (H+L) Secondary Antibody [DyLight 594] (Pre-adsorbed) heavy gamma chains on mouse IgG and light chains on all mouse immunoglobulins. This antibody has been pre-adsorbed against bovine, chicken, goat, guinea pig, hamster, horse, human, rabbit, rat or sheep IgG

Clonality

Polyclonal

Host

Donkey

Isotype

IgG

Description

Affinity purified antibody is > 95% based on SDS-PAGE. Donkey serum was obtained from healthy animals of US origin and under the care of a registered veterinarian. DyLight 594 (Ex = 593 nm; Em = 618 nm). After reconstituted, prepare fresh working dilution daily

Product is stable for up to 4 weeks at 2-8C after rehydration. For extended storage after rehydration, add an equal volume of glycerol and store at -20C.

Scientific Data Images

Donkey anti-Mouse IgG (H+L) Secondary Antibody [DyLight 594] (Pre-adsorbed)

Immunocytochemistry/ Immunofluorescence: Donkey anti-Mouse IgG (H+L) Secondary Antibody [DyLight 594] (Pre-adsorbed) [NBP1-75617] -

Immunocytochemistry/ Immunofluorescence: Donkey anti-Mouse IgG (H+L) Secondary Antibody [DyLight 594] (Pre-adsorbed) [NBP1-75617] - Effect of varied WAT masses from obese mice on vimentin & CD24 protein expression in MDA-MB-231 cells. (a) Representative images of MDA-MB-231 cells cultured with or without 200 mg or 800 mg of obese subcutaneous or visceral WAT-CM & co-stained with anti-vimentin, anti-CD24, & DAPI; scale bar = 100 µm. (b) & (c) are the relative expression of the respective markers, mean ± SEM (n = 6 biological replicates). The average protein expression from five images per replicate were normalized to DAPI & analyzed relative to the control cultures. No groups were found to be significantly different by the Friedman test. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32899433), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Donkey anti-Mouse IgG (H+L) Secondary Antibody [DyLight 594] (Pre-adsorbed)

Immunocytochemistry/ Immunofluorescence: Donkey anti-Mouse IgG (H+L) Secondary Antibody [DyLight 594] (Pre-adsorbed) [NBP1-75617] -

Immunocytochemistry/ Immunofluorescence: Donkey anti-Mouse IgG (H+L) Secondary Antibody [DyLight 594] (Pre-adsorbed) [NBP1-75617] - Mouse WAT-CM induces the expression of CD24 in mesenchymal MDA-MB-231 cells but does not affect the expression of vimentin. Representative images of MDA-MB-231 cells cultured with or without lean or obese subcutaneous or visceral WAT-CM & co-stained for vimentin, CD24, & DAPI (a); scale bar = 100 µM. (b) & (c) are the relative expression of the respective markers, mean ± SEM (n = 6 biological replicates). The average protein expression from five images per replicate were normalized to DAPI & analyzed relative to the control cultures. Significance was determined by Friedman’s test. If significant, Dunn’s test was used for further pairwise comparison; * p < 0.05. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32899433), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Donkey anti-Mouse IgG (H+L) Secondary Antibody [DyLight 594] (Pre-adsorbed)

Immunocytochemistry/ Immunofluorescence: Donkey anti-Mouse IgG (H+L) Secondary Antibody [DyLight 594] (Pre-adsorbed) [NBP1-75617] -

Immunocytochemistry/ Immunofluorescence: Donkey anti-Mouse IgG (H+L) Secondary Antibody [DyLight 594] (Pre-adsorbed) [NBP1-75617] - Mouse WAT-CM induces the expression of epithelial biomarkers in mesenchymal triple negative breast cancer cell line grown in a 3D culture. (a) Representative images of MDA-MB-231 cells cultured with or without lean or obese subcutaneous or visceral WAT-CM & co-stained for E-cadherin, claudin-7, & DAPI; scale bar = 100 µm. (b) & (c) are the relative expression of the respective markers, mean ± SEM (n = 6 biological replicates). The average protein expression from five images per replicate were normalized to DAPI & analyzed relative to the control cultures. Significance was determined by the Friedman test. If significant, Dunn’s test was used for further pairwise comparison; * p < 0.05. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32899433), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications

Application
Recommended Usage

Flow Cytometry

1:20 - 1:2000

Immunomicroscopy

1:20 - 1:2000
Application Notes
This conjugate is suitable for immunomicroscopy and flow cytometry.

Formulation, Preparation, and Storage

Purification

Affinity purified

Reconstitution

Rehydrate with 1.1 ml of deionized water and let stand 30 minutes at room temperature to dissolve. (Product has been overfilled to ensure complete recovery.) Centrifuge to remove any particulates. Prepare fresh working dilution daily.

Formulation

10 mM Sodium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 1 % (w/v) BSA, Protease/IgG free

Format

Pre-adsorbed

Preservative

0.05% 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. Mix reconstituted liquid with an equal volume of glycerol, aliquot contents and freeze at -20C or below for long term storage.

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

Certificate of Analysis

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

Product Specific Notices



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.

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