Cell Signaling Technology

Product Pathways - Companion Products

Rabbit IgG Isotype Control (Alexa Fluor® 647 Conjugate) #3452

Applications Reactivity Sensitivity Source
F Endogenous Rabbit

Applications Key:  F=Flow Cytometry
Reactivity Key:
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

The purified antibody is not directed against any known antigen. It functions as an isotype control for rabbit antibodies.

Source / Purification

The antibody was isolated from naive rabbit sera and purified by affinity chromotography. The antibody was conjugated to Alexa Fluor® 647 under optimal conditions with a F/P ratio of 2-6.Important Note on Use: This control antibody must be diluted to the same concentration (not dilution) as the specific antibody in analysis. Higher background fluorescence may result if excessive amounts of rabbit IgG isotype control are used. Do not use this antibody at 1:50 dilution in the same way as recommended for other CST Alexa Fluor® conjugated antibodies.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, untreated (green) or etoposide-treated (blue), using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb (Alexa Fluor® 647 Conjugate) #9720 compared to Rabbit IgG Isotype Control (Alexa Fluor® 647 Conjugate) (red).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, untreated (green) or LY294002- and wortmannin-treated (blue), using Phospho-Akt (Ser473) (193H12) Rabbit mAb (Alexa Fluor® 647 Conjugate) #2337 compared to Rabbit IgG Isotype Control (Alexa Fluor® 647 Conjugate) (red).

Description

Rabbit IgG was conjugated to Alexa Fluor® 647 fluorescent dye under optimal conditions, and tested in-house for direct flow cytometric analysis of human and mouse cells. Alexa Fluor® dye is maximally excited by red light (e.g. 633 nm He-Ne laser). Antibody conjugated of the Alexa Fluor® 647 dye produce far-red-fluorescence emission with a peak at 655 nm.

Directions for Use

Important! This control antibody must be diluted to the same concentration (not dilution) as the specific antibody in analysis. Higher background fluorescence may result if excessive amounts of rabbit IgG isotype control are used. Do not use this antibody at 1:50 dilution in the same way as recommended for other CST Alexa Fluor® conjugated antibodies.

Background

Isotype control antibodies are used to estimate the nonspecific binding of target primary antibodies due to Fc receptor binding or other protein-protein interactions. An isotype control antibody should have the same immunoglobulin type and be used at the same concentration as the test antibody.

Application References

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Companion Products

Alexa Fluor® is a registered trademark of Molecular Probes, Inc.


For Research Use Only. Not For Use In Diagnostic Procedures.

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