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

Images

 
Western Blot: Goat anti-Rabbit IgG (H+L) Secondary Antibody [DyLight 488] (Pre-adsorbed) [NBP1-72944] - Western Blot of Unconjugated Goat anti-Rabbit IgG (H+L) Secondary antibody [DyLight 488] (Pre-adsorbed). Lane M: 3 ...read more
Immunocytochemistry/ Immunofluorescence: Goat anti-Rabbit IgG (H+L) Secondary Antibody [DyLight 488] (Pre-adsorbed) [NBP1-72944] - Myosin expression in HeLa cells. Cells were fixed in 4% paraformaldehyde for 20 minutes ...read more
Immunocytochemistry/ Immunofluorescence: Goat anti-Rabbit IgG (H+L) Secondary Antibody [DyLight 488] (Pre-adsorbed) [NBP1-72944]
Immunocytochemistry/ Immunofluorescence: Goat anti-Rabbit IgG (H+L) Secondary Antibody [DyLight 488] (Pre-adsorbed) [NBP1-72944] - DyLight 488 conjugated anti-Rabbit IgG was used to demonstrate 2 color STED ...read more

Product Details

Summary
Reactivity RbSpecies Glossary
Applications WB, FLISA, ICC/IF
Clonality
Polyclonal
Host
Goat
Conjugate
DyLight 488
Format
Pre-adsorbed
Concentration
LYOPH

Order Details

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

Immunogen
Rabbit IgG whole molecule
Specificity
This antibody was pre-adsorbed against Bovine, Chicken, Goat, Guinea Pig, Hamster, Horse, Human, Mouse, Rat, and Sheep Serum Proteins.

No reaction was observed against Bovine, Chicken, Goat, Guinea Pig, Hamster, Horse, Human, Mouse, Rat and Sheep Serum Proteins. This antibody will react with heavy chains of rabbit IgG and with light chains of most rabbit immunoglobulins.
Isotype
IgG
Clonality
Polyclonal
Host
Goat
Purity
Multi-step

Applications/Dilutions

Dilutions
  • Fluorophore-linked immunosorbent assay 1:20000
  • Immunocytochemistry/Immunofluorescence 1:5000
  • Western Blot 1:10000-1:25000
Application Notes
This secondary antibody 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.
Reviewed Applications
Read 1 Review rated 5
using
NBP1-72944 in the following applications:

Packaging, Storage & Formulations

Storage
Store lyophilized antibody at 4C in the dark. Aliquot reconstituted liquid and store at -20C. Avoid freeze-thaw cycles.
Buffer
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
Preservative
0.01% Sodium Azide
Concentration
LYOPH
Purity
Multi-step
Reconstitution Instructions
Reconstitute with 100 ul deionized water (or equivalent).

Notes

This product was prepared from monospecific antiserum by immunoaffinity chromatography using Rabbit 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, Rabbit IgG and Rabbit Serum

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.

Background

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

Limitations

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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Review for IgG (H+L) Antibody (NBP1-72944) (1) 51

Average Rating: 5
(Based on 1 review)
We have 1 review tested in 1 species: Human.

Reviews using NBP1-72944:
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Immunofluorescence IgG (H+L) NBP1-72944
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5
reviewed by:
Verified Customer
IF Human 11/12/2014
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Summary

ApplicationImmunofluorescence
Sample Testedhela
SpeciesHuman

Blocking

Blocking Detailsblocked in 1% BSA in PBS, 20 mins at RT

Primary Anitbody

Dilution Ratioprimary antibody against myosin, diluted 1/500 in 1%BSA, incubated 20 min RT

Secondary Antibody

Secondary Descriptionthis is the secondary
Secondary Concentration1/500 in 1% BSA in PBS, 20 mins at RT

Details

Detection Notesimmunofluorescence
Fixation Detailsfixed in 4% paraformaldehyde, 20 mins at RT
Wash Descriptionwashed three times between each incubation with PBS

Product General Protocols

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Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for IgG (H+L) Antibody (NBP1-72944) (0)

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Verified Customer
11/12/2014
Application: IF
Species: Human