Donkey anti-Rabbit IgG (H+L) Secondary Antibody [DyLight 488] (Pre-adsorbed)

Novus Biologicals, part of Bio-Techne | Catalog # NBP1-75292

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NBP1-75292
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Key Product Details

Species Reactivity

Rabbit

Applications

Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Paraffin, Immunomicroscopy

Label

DyLight 488 (Excitation = 493 nm, Emission = 518 nm)

Antibody Source

Polyclonal Donkey IgG

Format

Pre-adsorbed

Concentration

LYOPH mg/ml

View all IgG (H+L) Secondary Antibodies »

Product Specifications

Immunogen

This Donkey anti-Rabbit IgG (H+L) Secondary Antibody [DyLight 488] (Pre-adsorbed) was developed against purified rabbit IgG (H&L).

Reactivity Notes

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

Specificity

Based on IEP, this Donkey anti-Rabbit IgG (H+L) Secondary Antibody [DyLight 488] (Pre-adsorbed) heavy gamma chains on rabbit IgG and light chains on all rabbit immunoglobulins. This antibody has been pre-adsorbed against bovine, chicken, goat, guinea pig, hamster, horse, human, mouse, rat or sheep IgG

Clonality

Polyclonal

Host

Donkey

Isotype

IgG

Description

Purity > 95% based on SDS-PAGE. Donkey serum was obtained from healthy animals of US origin and under the care of a registered veterinarian. 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

Immunocytochemistry/Immunofluorescence: Donkey anti-Rabbit IgG (H+L) Secondary Antibody [DyLight 488] (Pre-adsorbed) [NBP1-75292] - Brain endothelial cells were stained with anti-CD31 antibody (raised in rabbit) and then with NBP1-75292. After counterstaining with DAPI, image was acquired with confocal microscopy. Antibody dilution: 1:1000 for 2 hours at r.t. Image from verified customer review.

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.

Use in Immunohistochemistry (PMID: 28821924) and Immunohistochemistry-Paraffin (PMID: 31882863) was reported in the scientific literature. Donkey anti-Rabbit IgG (H+L) Secondary Antibody [DyLight 488] (Pre-adsorbed) validated for Immunocytochemistry/Immunofluorescence from a verified customer review.

Reviewed Applications

Read 1 review rated 5 using NBP1-75292 in the following applications:

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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

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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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