Invitrogen™ Armenian Hamster IgG Isotype Control (eBio299Arm), eFluor™ 660, eBioscience™

Catalog No.	Quantity
50-112-4376	100 μg

Catalog No. 50-112-4376 Supplier Invitrogen™ Supplier No. 50488882

Description

Armenian Hamster Monoclonal Isotype Control

The eBio299Arm monoclonal antibody is useful as an isotype control immunoglobulin. Applications Reported: This eBio299Arm antibody has been reported for use in immunocytochemistry, immunohistochemistry, and flow cytometric analysis. Applications Tested: This eBio299Arm antibody has been tested by flow cytometric analysis of normal human peripheral blood cells and mouse splenocytes. Use isotype control at the same concentration as experimental antibody. eFluor 660 is a replacement for Alexa Fluor 647. eFluor 660 emits at 659 nm and is excited with the red laser (633 nm). Please make sure that your instrument is capable of detecting this fluorochome. Excitation: 633-647 nm; Emission: 668 nm; Laser: Red Laser. Filtration: 0.2 μm post-manufacturing filtered.

The isotype of a primary antibody and the application it is being used in can result in background staining. Primary antibody background noise can be caused by binding to Fc receptors on target cells; by non-specific interactions with cellular proteins, carbohydrates, and lipids; or by cell autofluorescence. Isotype control antibodies can act as negative controls to help differentiate non-specific background signal from specific antibody signal because they have no relevant specificity to a target antigen. While isotype controls are most commonly used in flow cytometry, they are useful in other applications such as chromatin immunoprecipitation (ChIP), immunohistochemistry, and gel shifts. Isotype controls should match with the primary antibody species and isotype so that the level of specific staining by the primary antibody may be accurately determined. If using directly labeled primary antibodies, the isotype control works best if conjugated with the same label as the test antibody.

Specifications

• Antigen: Armenian Hamster IgG
• Applications: Control, Flow Cytometry, Immunohistochemistry, Immunocytochemistry, Immunohistochemistry
• Classification: Monoclonal
• Clone: eBio299Arm
• Concentration: 0.2 mg/mL
• Conjugate: eFluor 660
• Formulation: PBS with 0.09% sodium azide; pH 7.2
• Gene Alias: IgG; Immunoglobulin G; ImmunoglobulinG
• Host Species: Armenian Hamster
• Purification Method: Affinity chromatography
• Quantity: 100 μg
• Regulatory Status: RUO
• Primary or Secondary: Isotype Control (Primary)
• Target Species: Not Applicable
• Content And Storage: 4°C, store in dark, DO NOT FREEZE!
• Product Type: Isotype Control
• Form: Liquid
• Isotype: IgG

Frequently Asked Questions (FAQs)

I am unable to analyze my cells stained with eFluor Organic Dyes today. What options do I have?

Our options will depend on the samples you are analyzing.

If cell viability is not critical, you can store your stained samples at 4 degrees C or on ice overnight in the dark and analyze the following day.

For samples stained with eFluor organic fluorochromes, we recommend that cells be suspended in 100 uL of Flow Cytometry Staining Buffer (Cat. No. 00-4222) and 100 uL of eBioscience IC Fixation Buffer (Cat. No. 00-8222); samples can be incubated for up to 3 days at 4 degrees C in the dark. Alternatively, the 1-step Fix/Lyse Solution (Cat. No. 00-5333) can be used. This is a great option when working with whole blood but also works for other cell types.

Can the eFluor Organic fluorochromes be used for intracellular staining?

Yes, the eFluor Organic fluorochromes can be used for intracellular staining. The eFluor organic fluorochromes maintain bright signal and require minimal changes in compensation when fixed with eBioscience IC Fixation Buffer (Cat. No. 00-8222-49) and Permeabilization Buffer (Cat. No. 00-8333-56) or 1-step Fix/Lyse Solution (Cat. No. 00-5333-54, 00-5333-57) (as compared to live cells).

Is the eFluor 660 fluorochrome compatible with Anti-Cy5/Alexa Fluor 647 beads?

Yes, in-house studies have demonstrated that the eFluor 660 fluorochrome is recognized by Anti-Cy5/Alexa Fluor 647 beads. Side by side studies with Alexa Fluor 647 versus eFluor 660 conjugated antibodies have demonstrated comparable results.

Are the eFluor Organic Dyes photo-labile?

As with other fluorochromes, we recommend minimal exposure to light to maintain optimal signal.