Key Product Details

Species Reactivity

Goat

Applications

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

Label

Unconjugated

Antibody Source

Polyclonal Donkey IgG

Format

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

Product Specifications

Immunogen

Goat IgG whole molecule

Specificity

This antibody was pre-adsorbed against Chicken, Guinea Pig, Hamster, Horse, Mouse, Rabbit, and Rat Serum Proteins. No reaction was observed against Chicken, Guinea Pig, Hamster, Horse, Mouse, Rabbit or Rat Serum Proteins. Specificity was confirmed using ELISA minimal cross reactivity against the targets stated above.

Clonality

Polyclonal

Host

Donkey

Isotype

IgG

Description

This product was prepared from monospecific antiserum by immunoaffinity chromatography using Goat 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, Goat IgG and Goat 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-Goat IgG (H+L) Secondary Antibody (Pre-adsorbed)

Western Blot: Donkey anti-Goat IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP2-60657]

Western Blot: Donkey anti-Goat IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP2-60657]

Western Blot: Donkey anti-Goat IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP2-60657] - Lane 1: Goat IgG Load: 50 ug per lane Secondary antibody: Goat IgG (H&L) Antibody DyLight (TM) 488 conjugated Pre-Adsorbed at 1:1,000 for 60 min at RT Block for30 min at RT Predicted/Observed size: 55 and 28 kDa, 55 and 28 kDa Image using the DyLight 488 form of this antibody.
Immunocytochemistry/ Immunofluorescence: Donkey anti-Goat IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP2-60657]

Immunocytochemistry/ Immunofluorescence: Donkey anti-Goat IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP2-60657]

Immunocytochemistry/Immunofluorescence: Donkey anti-Goat IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP2-60657] - Image shows detection of PD-1 in mouse frozen tissue sections (salivary gland injected with human T cells) using DyLight™ 488 conjugated donkey anti-goat IgG (H+L) secondary antibody (green). Image from verified customer review. Image using the DyLight 488 form of this antibody.
Western Blot: Donkey anti-Goat IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP2-60657]

Western Blot: Donkey anti-Goat IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP2-60657]

Western Blot: Donkey anti-Goat IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP2-60657] - Lane 1: Goat IgG. Lane 2: None. Load: 50 ng per lane. Primary antibody: None. Secondary antibody: Texas Red donkey secondary antibody at 1:1,000 for 60 min at RT. Blocking: incubated with blocking buffer for 30 min at RT. Predicted/Observed size: 25 & 55 kDa, 25 & 55 kDa for Goat IgG. Other band(s): None. Image using the Texas Red form of this antibody.
Western Blot: Donkey anti-Goat IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP2-60657]

Western Blot: Donkey anti-Goat IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP2-60657]

Western Blot: Donkey anti-Goat IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP2-60657] - Rabbit anti-Transferrin, Goat-anti-Alpha-1-Anti-Trypsin, and Mouse-a-GST. Detection shown was using DyLight549 Donkey anti-Rabbit IgG, DyLight 488 Donkey anti-Mouse IgG, and DyLight 649 Donkey anti-Goat IgG at 1:10000 (in blocking buffer min RT). Blots were washed, rinsed in methanol and dried. Image using the DyLight 649 form of this antibody.
Immunocytochemistry/ Immunofluorescence: Donkey anti-Goat IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP2-60657]

Immunocytochemistry/ Immunofluorescence: Donkey anti-Goat IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP2-60657]

Immunocytochemistry/Immunofluorescence: Donkey anti-Goat IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP2-60657] - Image shows anti-Histone detection using a DyLight™ 488 conjugate (green). Anti-Tubulin was detected using a DyLight™ 549 conjugate (red). Nuclei were counter-stained using DAPI (blue). Image using the DyLight 488 form of this antibody.
Fluorophore-linked immunosorbent assay: Donkey anti-Goat IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP2-60657]

Fluorophore-linked immunosorbent assay: Donkey anti-Goat IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP2-60657]

Fluorophore-linked immunosorbent assay: Donkey anti-Goat IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP2-60657] - Using the DyLight 405 format of this antibody.
ELISA: Donkey anti-Goat IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP2-60657]

ELISA: Donkey anti-Goat IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP2-60657]

ELISA: Donkey anti-Goat IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP2-60657] - ELISA Results of Donkey Donkey anti-Goat IgG (H+L) Secondary antibody (Pre-adsorbed) tested against purified Goat IgG MX7. Each well was coated in duplicate with 1.0 ug of Goat IgG as well as Chicken IgG, Guinea Porcine IgG, Hamster IgG, Equine IgG, Mouse IgG, Rabbit IgG, and Rat IgG. The working dilution is 1:56,000. 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 Mx7 HRP conjugated and TMB substrate.

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

Application
Recommended Usage

ELISA

1:20000 - 1:100000

Immunohistochemistry

1:200-1:1000

Western Blot

1:500-1:2000
Application Notes
This product has been tested by ELISA and is suitable for western blot and immunohistochemistry, as well as other assays requiring lot-to-lot consistency.

Flow Cytometry Panel Builder

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Advanced Features

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Flow Cytometry

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

Certificate of Analysis

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

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Protocols

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FAQs

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