Key Product Details

Species Reactivity

Mouse

Applications

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, ELISA, Immunocytochemistry/ Immunofluorescence, Chromatin Immunoprecipitation (ChIP), Dot Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

Format

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

Product Specifications

Immunogen

Mouse IgG whole molecule

Specificity

This antibody was pre-adsorbed against Human Serum Proteins. No reaction was observed against Human Serum Proteins. Specificity was confirmed using ELISA at less than 1% cross reactivity against Human IgG.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Description

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

Western Blot: Goat anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72739]

Western Blot: Goat anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72739]

Western Blot: Goat anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72739] - 100 ug of Mouse IgG (Lane 2) was run on a 4-20% gel and transferred to 0.45 um nitrocellulose. After blocking with 1% BSA-TTBS 30 min at 20C, Antibody was used at 1:1000 in blocking buffer for Fluorescent Western blotting. Molecular weight markers are in lane 1. Image from the DyLight 488 version of this antibody.
Immunocytochemistry/ Immunofluorescence: Goat anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72739]

Immunocytochemistry/ Immunofluorescence: Goat anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72739]

Immunocytochemistry/Immunofluorescence: Goat anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72739] - DyLight and ATTO dye conjugated antibodies provide high signal and low background for confocal microscopy and high resolution Stimulated Emission Depletion (STED) Microscopy. Both Dylight and Atto conjugated secondary antibodies maintained robust, intense signal during repeated laser excitation and de-excitation used during STED microscopy. Shown here are: A. (Green) Mouse anti NuP (NuP=Nuclear Pore Protein) detected with Dylight 488 Goat anti mouse B. (Red) Rabbit Anti Ezh1/2 Pab (Ezh=enhancer of zeste homology) with detection by ATTO 425 conjugated Goat anti Rabbit C. (Red and Green) Images combined. Image from the DyLight 488 version of this antibody.
Immunohistochemistry-Paraffin: Goat anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72739]

Immunohistochemistry-Paraffin: Goat anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72739]

Immunohistochemistry-Paraffin: Goat anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72739] - Staining of paraffin embedded mouse tissue. Goat anti-Mouse DyLight 488 was used as a secondary antibody. Image from verified customer review. Image from the DyLight 488 version of this antibody.
Immunocytochemistry/ Immunofluorescence: Goat anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72739]

Immunocytochemistry/ Immunofluorescence: Goat anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72739]

Immunocytochemistry/Immunofluorescence: Goat anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72739] - DyLight and ATTO dye conjugated antibodies provide high signal and low background for confocal microscopy (upper images) and high resolution Stimulated Emission Depletion (STED) Microscopy (lower images). Both Dylight and ATTO conjugated secondary antibodies maintained robust, intense signal during repeated laser excitation and de-excitation used during STED microscopy. Shown here are: A. (Green) Mouse anti NuP (NuP=Nuclear Pore Protein) detected with Dylight 488 Goat anti mouse B. (Red) Rabbit Anti Ezh1/2 Pab (Ezh=enhancer of zeste homology) with detection by ATTO 425 conjugated Goat anti Rabbit (Red and Green) Images combined. Image from the DyLight 488 version of this antibody.
Immunohistochemistry-Frozen: Goat anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72739]

Immunohistochemistry-Frozen: Goat anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72739]

Immunohistochemistry-Frozen: Goat anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72739] - MDA-MB-231 cells were stained. Goat anti-Mouse DyLight 488 was used as a secondary antibody and DRAQ5 was used for nuclear stain. Image from verified customer review. Image from the DyLight 488 version of this antibody.
Dot Blot: Goat anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72739] - Dot Blot of Goat anti-Mouse IgG antibody Alkaline Phosphatase Conjugated Min X Bv Ch Gt GP Ham Hs Hu Rb, Rt, Sh Serum Proteins. Antigen: Mouse IgG. Load: 200 ng, 66.7 ng, 22.2 ng, 7.41 ng, or 2.47 ng as indicated.Primary antibody: Anti-Mouse IgG antibody Alkaline Phosphatase Conjugated at 1:1,000 for 60 min at 4C.Secondary antibody: N/A.Block: Blocking Buffer for Fluorescent Western Blotting for 60 min at RT. Image using the Alkaline Phosphatase format of this antibody.
Dot Blot: Goat anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72739] - A three-fold serial dilution of Mouse IgG starting at 200ug was spotted onto 0.45 um nitrocellulose. After blocking in 5% Blotto 1 Hour at 20C, Antibody was used at 1:1000 in blocking buffer for Fluorescent Western blotting. Image using the FITC format of this antibody.
ELISA: Goat anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72739]

ELISA: Goat anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72739]

ELISA: Goat anti-Mouse IgG (H+L) Secondary Antibody (Pre-adsorbed) [NBP1-72739] - ELISA results of purified Goat anti-Mouse IgG (H+L) Secondary Antibody [Unconjugated] (Pre-adsorbed) tested against purified Mouse IgG. Each well was coated in duplicate with 1.0 ug of Mouse IgG red line) Human IgG (green line). 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 blocking buffer, Gt-anti-Donkey HRP conjugate 616-1302, and TMB substrate.

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

Application
Recommended Usage

ELISA

1:100000 - 1:500000

Immunohistochemistry

1:1000 - 1:6000

Immunohistochemistry-Frozen

1:5000 - 1:25000

Immunohistochemistry-Paraffin

1:5000 - 1:25000

Western Blot

1:3000 - 1:12000
Application Notes
This product has been tested by ELISA and is suitable for ELISA, western blot, and immunohistochemistry, as well as other assays requiring lot-to-lot consistency.

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

Certificate of Analysis

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

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Protocols

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FAQs

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