Invitrogen™ F(ab')2-Goat anti-Rat IgG (H+L) Secondary Antibody, eFluor™ 660, eBioscience™

Catalog No.	Quantity
50-112-4350	100 μg

Catalog No. 50-112-4350 Supplier Invitrogen™ Supplier No. 50401782

Description

F(ab')2-Goat Polyclonal Secondary Antibody

Description: The goat anti-rat IgG polyclonal antibody reacts with the heavy and light chains of rat IgG, with minimal cross reactivity to human, mouse, bovine, goat, hamster, and rabbit serum proteins. Reactivity has been shown to all IgG subclasses: IgG1, IgG2a, IgG2b, IgG2c and some crossreactivity to rat IgM. The antibody is a F(ab')2 fragment which contains the 2 Fab domains still linked through the hinge region of the immunoglobulin. Removal of the Fc region helps to reduce issues typically found with Fc-mediated binding. This polyclonal has been validated as a secondary reagent (its ability to recognize cells stained with a rat IgG followed by this polyclonal antibody). Applications Reported: This poly antibody has been reported for use in flow cytometric analysis. Applications Tested: This poly antibody has been tested by flow cytometric analysis of mouse splenocytes first stained with rat anti-mouse antibody. This can be used at less than or equal to 0.25 μg per test. A test is defined as the amount (μg) of antibody that will stain a cell sample in a final volume of 100 μL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. 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 dete...

Anti-Rat secondary antibodies are affinity-purified antibodies with well-characterized specificity for rat immunoglobulins and are useful in the detection, sorting or purification of its specified target. Secondary antibodies offer increased versatility enabling users to use many detection systems (e.g. HRP, AP, fluorescence). They can also provide greater sensitivity through signal amplification as multiple secondary antibodies can bind to a single primary antibody. Most commonly, secondary antibodies are generated by immunizing the host animal with a pooled population of immunoglobulins from the target species and can be further purified and modified (i.e. immunoaffinity chromatography, antibody fragmentation, label conjugation, etc.) to generate highly specific reagents.

Specifications

• Antigen: Rat IgG (H+L)
• Applications: Flow Cytometry
• Classification: Polyclonal
• Concentration: 0.2 mg/mL
• Conjugate: eFluor 660
• Formulation: PBS with 0.09% sodium azide; pH 7.2
• Host Species: Goat
• Purification Method: Affinity chromatography
• Quantity: 100 μg
• Regulatory Status: RUO
• Primary or Secondary: Secondary
• Target Species: Rat
• Content And Storage: 4°C, store in dark, DO NOT FREEZE!
• Product Type: Secondary Antibody
• Form: Liquid
• Isotype: Ig

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.

What buffers are compatible with the eFluor Organic fluorochromes and eVolve QDots?

The eFluor Organic fluorochromes and eVolve QDots can be used with flow staining buffers containing PBS and protein.

What is the difference between eFluor Organic Dyes and eVolve Qdots?

The eFluor Organic Dyes (eFluor 450, APC-eFluor 780, PerCP-eFluor 710, eFluor 710) are conventional fluorochromes. In contrast, the eVolve line of products are Quantum dots.

Are the eFluor Organic Dyes photo-labile?

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

Safety and Handling

WARNING: Reproductive Harm - www.P65Warnings.ca.gov