Rabbit anti-Rat IgG (H+L) Secondary Antibody [DyLight 680] (Pre-adsorbed)

Novus Biologicals, part of Bio-Techne | Catalog # NBP1-72991

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NBP1-72991
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Key Product Details

Species Reactivity

Rat

Applications

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

Label

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

Antibody Source

Polyclonal Rabbit IgG

Format

Pre-adsorbed

Concentration

LYOPH mg/ml

View all IgG (H+L) Secondary Antibodies »

Product Specifications

Immunogen

Rat IgG whole molecule

Specificity

This antibody was pre-adsorbed against Human Serum Proteins. No reaction was observed against Human Serum Proteins. This antibody will react with heavy chains of Rat IgG and with light chains of most Rat immunoglobulins.

Clonality

Polyclonal

Host

Rabbit

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 Rat 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-Rabbit Serum, Rat IgG and Rat Serum

Scientific Data Images

Western Blot: Rabbit anti-Rat IgG (H+L) Secondary Antibody [DyLight 680] (Pre-adsorbed) [NBP1-72991] - Western Blot using anti-Rat IgG (H+L) Secondary Antibody [DyLight 680] (Pre-absorbed). Lane M: 3 ul Molecular Ladder. Lane 1: Rat IgG Isotype Control (NBP1-97040). Lane 2: Rat IgG, Fc Isotype Control (NBP1-96967). Lane 3: Rat IgG Fab Fragment (NBP1-97014). Lane 4: Rat IgM Isotype Control (NBP1-96776). Lane 5: Rat Serum. All samples were reduced. Load: 50 ng per lane. Blocking for 30 min at RT. Primary Antibody: Anti-Rat IgG (H+L) Antibody, blocked for 60 min at RT. Secondary Antibody: Anti-Rabbit IgG Peroxidase Conjugated Antibody blocked for 30 min at RT. Predicted/Observed Size: 25 and 55 kDa for Rat IgG and Serum, 25 kDa for F(c) and Fab, 78 and 25 kDa for IgM. Rat F(c) migrates slightly higher.
Western Blot: Rabbit anti-Rat IgG (H+L) Secondary Antibody [DyLight 680] (Pre-adsorbed) [NBP1-72991] - Anti-tubulin was detected using a DyLight 680 conjugate. Anti-TNF alpha was detected using a DyLight 800 conjugate. The image was captured using the Odyssey Infrared Imaging System developed by LI-COR.
Rabbit anti-Rat IgG (H+L) Secondary Antibody [DyLight 680] (Pre-adsorbed)

Rabbit anti-Rat IgG (H+L) Secondary Antibody [DyLight 680] (Pre-adsorbed)

Western Blot of Rabbit anti-Rat IgG (H+L) Secondary Antibody [DyLight 680] (Pre-adsorbed)
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Applications

Application
Recommended Usage

ELISA

1:20000

Immunocytochemistry/ Immunofluorescence

1:5000

Western Blot

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. The emission spectra for this DyLight(TM) conjugate match the principle output wavelengths of most common fluorescence instrumentation.

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

Format

Pre-adsorbed

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.

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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