Invitrogen™ CD275 (B7-H2) Monoclonal Antibody (HK5.3), eFluor™ 660, eBioscience™

Catalog No.	Quantity
50-112-4415	100 μg

Catalog No. 50-112-4415 Supplier Invitrogen™ Supplier No. 50598582

Description

Rat Monoclonal Antibody

The HK5.3 monoclonal antibody reacts with mouse B7RP-1, also known as B7h, B7-H2, GL50 and ICOS Ligand. B7RP-1, a member of the B7 family, has a predicted molecular weight of ~40 kDa and belongs to the Ig superfamily. Mouse B7RP-1 is expressed by antigen presenting cells including B cells, monocytes/macrophages and dendritic cells. B7RP-1 binds to the ICOS molecule (AILIM, CRP-1) expressed by activated T cells. The interaction of ICOS/B7RP-1 plays an important role in the T cell costimulation pathway. Applications Reported: This HK5.3 antibody has been reported for use in flow cytometric analysis. Applications Tested: This HK5.3 antibody has been tested by flow cytometric analysis of mouse splenocytes. This can be used at less than or equal to 0.5 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 detecting this fluorochome. Excitation: 633-647 nm; Emission: 668 nm; Laser: Red Laser. Filtration: 0.2 μm post-manufacturing filtered.

Inducible co-stimulator ligand (ICOSL) is a specific ligand on antigen-presenting cells and cells of the peripheral tissue that binds to the inducible co-stimulator receptor (ICOS). ICOS belongs to the CD28/CD152 receptor family that regulates T-cell activation and function. ICOSL is expressed on monocytes, dendritic cells and B cells and can be induced by inflammatory stimuli in peripheral tissue. Binding to ICOSL delivers a co-stimulatory signal for T cell proliferation and cytokine secretion.

Specifications

• Antigen: CD275 (B7-H2)
• Applications: Flow Cytometry
• Classification: Monoclonal
• Clone: HK5.3
• Concentration: 0.2 mg/mL
• Conjugate: eFluor 660
• Formulation: PBS with 0.09% sodium azide; pH 7.2
• Gene: Icosl
• Gene Accession No.: Q9JHJ8
• Gene Alias: AU044799; B7 homolog 2; B7 homologue 2; B7h; B7H2; B7-H2; B7-like protein Gl50; B7-related protein 1; B7RP1; B7RP-1; BG071784; CD275; GI50; GL50; GL50-B; ICOS ligand; ICOSL; ICOS-L; ICOSLG; inducible T-cell costimulator ligand; inducible T-cell co-stimulator ligand; KIAA0653; LICOS; Ly115l; mKIAA0653; RGD1562791; transmembrane protein B7-H2 ICOS ligand
• Gene Symbols: Icosl
• Host Species: Rat
• Purification Method: Affinity chromatography
• Quantity: 100 μg
• Regulatory Status: RUO
• Primary or Secondary: Primary
• Gene ID (Entrez): 50723
• Target Species: Mouse
• Content And Storage: 4°C, store in dark, DO NOT FREEZE!
• Product Type: Antibody
• Form: Liquid
• Isotype: IgG2a κ

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 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.