Goat anti-Rabbit IgG (H+L) Secondary Antibody [Rhodamine]

Novus Biologicals, part of Bio-Techne | Catalog # NB7575

Novus Biologicals, part of Bio-Techne

Key Product Details

Species Reactivity

Rabbit

Applications

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

Label

Rhodamine (Excitation = 570 nm, Emission = 590 nm)

Antibody Source

Polyclonal Goat IgG

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

View all IgG (H+L) Secondary Antibodies »

Product Specifications

Immunogen

Rabbit IgG whole molecule

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Description

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

This product was prepared from monospecific antiserum by immunoaffinity chromatography using Rabbit IgG coupled to agarose. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Goat Serum, Rabbit IgG and Rabbit Serum.

Scientific Data Images for Goat anti-Rabbit IgG (H+L) Secondary Antibody [Rhodamine]

Western Blot: Goat anti-Rabbit IgG (H+L) Secondary Antibody [Rhodamine] [NB7575] - Western Blot of Goat anti-Rabbit IgG (H+L) Secondary Antibody [Rhodamine]. Lane 1: Rabbit IgG. Lane 2: None. Load: 50 ng per lane.Primary antibody: None.Secondary antibody: Goat anti-Rabbit IgG (H+L) Secondary Antibody [Rhodamine] at 1:1000 for 60 min at RT.Block: Fluorescent blocking buffer for 30 min at RT.Predicted/Observed size: 25 & 55 kDa, 25 & 55 kDa for Rabbit IgG. Other band(s): None.
Western Blot: Goat anti-Rabbit IgG (H+L) Secondary Antibody [Rhodamine] [NB7575] - Lane 1: Rabbit IgG whole molecule. Lane 2: Rabbit IgG F(ab) Fragment. Lane 3: Rabbit IgG F(c) Fragment. Lane 4: Rabbit IgM Whole Molecule. Lane 5: Normal Rabbit Serum. All samples were reduced. Load: 50 ng per lane. Block: incubated with blocking buffer for 30 min at RT. Primary Antibody: Anti-Rabbit IgG (H&L) (GOAT) Antibody 1:1,000 for 60 min at RT. Secondary antibody: Anti-Goat IgG (DONKEY) Peroxidase Conjugated Antibody 1:40,000 in incubated with blocking buffer for 30 min at RT. Predicted/Observed Size: 25 and 50 kDa for Rabbit IgG and Serum, 25 kDa for F(c) and F(ab), 70 and 23 kDa for IgM. Rabbit F(c) migrates slightly higher.
Dot Blot: Goat anti-Rabbit IgG (H+L) Secondary Antibody [Rhodamine] [NB7575] - Dot Blot of Goat anti-Rabbit IgG (H+L) Secondary Antibody [Rhodamine].
Antigen: Rabbit IgG.
Load: Lane 1 - 50ng Lane 2 - 16.67ng Lane 3 - 5.56ng Lane 4 - 1.85ng Lane 5 - 0.62ng.
Primary antibody: none.
Secondary antibody: Goat anti-Rabbit IgG (H+L) Secondary Antibody [Rhodamine] at 1:1000 for 60 min at RT.
Block: Fluorescent blocking buffer for 60 min at RT.
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Applications for Goat anti-Rabbit IgG (H+L) Secondary Antibody [Rhodamine]

Application
Recommended Usage

Flow Cytometry

1:500 - 1:2500

Fluorophore-linked immunosorbent assay

1:10000 - 1:50000

Immunocytochemistry/ Immunofluorescence

1:1000 - 1:5000

Western Blot

1:1000 - 1:10000
Application Notes
This product has been tested by dot blot and 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.

Use in ICC/IF reported in secitific publication PMID: 32705217

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

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

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-Rabbit IgG (H+L) Secondary Antibody [Rhodamine]

Product Datasheets

Certificate of Analysis

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Product Specific Notices for Goat anti-Rabbit IgG (H+L) Secondary Antibody [Rhodamine]

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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