Key Product Details

Species Reactivity

Sheep

Applications

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

Label

Unconjugated

Antibody Source

Polyclonal Donkey IgG

Format

Pre-adsorbed
View all IgG (H+L) Secondary Antibodies »

Product Specifications

Immunogen

Donkey anti-Sheep IgG (H+L) Secondary Antibody (Pre-adsorbed) was produced by repeated immunization with Sheep IgG whole molecule in donkey.

Specificity

This antibody was pre-adsorbed against Chicken, Guinea Pig, Hamster, Horse, Human, Mouse, Rabbit, and Rat Serum Proteins. No reaction was observed against Chicken, Guinea Pig, Hamster, Horse, Human, Mouse, Rabbit and Rat Serum Proteins.

Clonality

Polyclonal

Host

Donkey

Isotype

IgG

Description

This product was prepared from monospecific antiserum by immunoaffinity chromatography using Sheep 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-Donkey Serum, Sheep IgG and Sheep Serum

Store vial at 4C prior to opening. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use. For extended storage aliquot contents and freeze at -20C or below. Avoid cycles of freezing and thawing.

Scientific Data Images for Donkey anti-Sheep IgG (H+L) Secondary Antibody (Pre-adsorbed)

Western Blot: Donkey anti-Sheep IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72745]

Western Blot: Donkey anti-Sheep IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72745]

Western Blot: Donkey anti-Sheep IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72745] - 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.
Immunocytochemistry/ Immunofluorescence: Donkey anti-Sheep IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72745]

Immunocytochemistry/ Immunofluorescence: Donkey anti-Sheep IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72745]

Immunocytochemistry/Immunofluorescence: Donkey anti-Sheep IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72745] - Immunocytochemistry/Immunofluorescence: Donkey anti-Sheep IgG (H+L) Secondary Antibody [DyLight 488] (Pre-absorbed) [NBP1-73002] Image from the DyLight 488 version of this antibody.
ELISA: Donkey anti-Sheep IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72745]

ELISA: Donkey anti-Sheep IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72745]

ELISA: Donkey anti-Sheep IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72745] - ELISA results of purified Polyclonal Donkey anti-Sheep IgG (H+L) Secondary Antibody [Alkaline Phosphatase] (Pre-adsorbed) tested against Sheep IgG. Each well was coated in duplicate with 1.0ug of antigen. The starting dilution of antibody was 5ug/ml and the X-axis represents the Log10 of a 3-fold dilution. This titration is a 4-parameter curve fit where the IC50 is defined as the titer of the antibody. Assay performed using Buffer. Using the Alkaline Phosphatase format of this antibody.
Fluorophore-linked immunosorbent assay: Donkey anti-Sheep IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72745]

Fluorophore-linked immunosorbent assay: Donkey anti-Sheep IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72745]

Fluorophore-linked immunosorbent assay: Donkey anti-Sheep IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72745] - Using the DyLight 405 format of this antibody.
ELISA: Donkey anti-Sheep IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72745]

ELISA: Donkey anti-Sheep IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72745]

ELISA: Donkey anti-Sheep IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72745] - ELISA Results of Donkey anti-Sheep IgG (H+L) Secondary antibody (Pre-adsorbed) tested against purified Sheep IgG Mx8. Each well was coated in duplicate with 1.0 ug of Sheep IgG [Red Line], Chicken IgG [Green Line], Guinea Porcine IgG [Blue Line], Hamster IgG [Purple Line], Equine IgG, Human IgG, Mouse IgG, Rabbit IgG, and Rat IgG. The working dilution of Sheep IgG MX8 is 1:49,000. The starting dilution of antibody was 5ug/ml and the X-axis represents the Log10 of a 3-fold dilution. This titration is a 4-parameter curve fit where the IC50 is defined as the titer of the antibody. Assay performed using HRP Conjugate Stabilizer, Goat Anti-Donkey IgG HRP conjugated, and TMB substrate.

Applications for Donkey anti-Sheep IgG (H+L) Secondary Antibody (Pre-adsorbed)

Application
Recommended Usage

ELISA

1:20000 - 1:100000

Immunohistochemistry

1:1000 - 1:5000

Western Blot

1:2000 - 1:10000
Application Notes
This product has been tested by ELISA and is suitable for use in immunoelectrophoresis, western-blot, competitive western-blot, ELISA and competitive ELISA assays. Specific conditions for reactivity and signal detection should be optimized by the end user.

Formulation, Preparation, and Storage

Purification

Multi-step

Formulation

0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2

Format

Pre-adsorbed

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-Sheep IgG (H+L) Secondary Antibody (Pre-adsorbed)

Certificate of Analysis

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Product Specific Notices for Donkey anti-Sheep IgG (H+L) Secondary Antibody (Pre-adsorbed)

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-Sheep IgG (H+L) Secondary Antibody (Pre-adsorbed)

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Protocols

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FAQs

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