Invitrogen™ CD294 (CRTH2) Monoclonal Antibody (BM16), eFluor™ 660, eBioscience™

Catalog No.	Quantity
50-294-942	100 Tests

Catalog No. 50-294-942 Supplier Invitrogen™ Supplier No. 50294942

Description

Rat Monoclonal Antibody

Description: This BM16 monoclonal antibody reacts with human CD294, which is also known as G protein-coupled receptor-44 (GPR-44) or chemoattractant receptor expressed on Th2 cells (CRTH2). CD294 is expressed on Th2 cells, some CD8+ cells, eosinophils, and basophils. Moreover, CD294 binds prostaglandin D2, which promotes leukocyte chemotaxis during allergic responses. CD294 has been reported to mediate prostaglandin D2-mediated suppression of apoptosis of Th2 cells by activating the PI3K pathway. Applications Reported: This BM16 antibody has been reported for use in flow cytometric analysis. Applications Tested: This BM16 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 μL (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. 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.

CRTH2 (GPR44) is a chemoattractant receptor-homologous molecule expressed on Th2 cells that is an identified receptor for the prostaglandin D2 (PGD2) and prostaglandin D receptor. CRTH2 is expressed on Th2 cells and eosinophils and mediates chemotaxis of these cells to PGD2 and is thus thought to be a key receptor mediating eosinophil and Th2 recruitment during allergic responses. However, CRTH2-null mice showed enhanced eosinophil recruitment into the lung consistent with observations that the CRTH2-null mice produced significantly higher amounts of interleukin-5 (IL-5) and IL-3. This suggests that CRTH2 plays a nonredundant role in restricting eosinophilia and allergic response in vivo. CRTH2 is also a novel member of the G protein-coupled leukocyte chemoattractant receptor family, which is selectively expressed in Th2 but not Th1 lineage cells. CRTH2 has been reported to be expressed in the central nervous system, peripheral tissues, and in blood cells, particularly activated T helper-2 cells. Two transcripts have been isolated: a 3.5 kb transcript expressed primarily in the thalamus, frontal cortex, pons and hippocampus; and a 3.4 kb transcript expressed in fetal liver, leukocytes and thymus. CRTH2 is selectively expressed in vivo in an activated state of Th2 cells including allergen-responsive Th2 cells, suggesting its pivotal roles in ongoing Th2-type immune reactions. At least two different isoforms of CRTH2 are known to exist. Single nucleotide polymorphisms in the 3' UTR of the CRTH2 gene have been associated with asthma susceptibility.

Specifications

• Antigen: CD294 (CRTH2)
• Applications: Flow Cytometry
• Classification: Monoclonal
• Clone: BM16
• Concentration: 5 μL/Test
• Conjugate: eFluor 660
• Formulation: PBS with BSA and 0.09% sodium azide; pH 7.2
• Gene: PTGDR2
• Gene Accession No.: Q9Y5Y4
• Gene Alias: CD294; chemoattractant receptor homologous molecule expressed on T helper type 2 cells; chemoattractant receptor-homologous molecule expressed on TH2 cells; CRTH2; DL1R; DP2; G protein-coupled receptor 44; Gpr44; G-protein coupled receptor 44; Grp45; MGC130436; PGD2 receptor; prostaglandin D2 receptor 2; prostaglandin D2 receptor CRTH2; Ptgdr2; putative G-protein coupled receptor 44
• Gene Symbols: PTGDR2
• Host Species: Rat
• Purification Method: Affinity chromatography
• Quantity: 100 Tests
• Regulatory Status: RUO
• Primary or Secondary: Primary
• Gene ID (Entrez): 11251
• Target Species: Human
• 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.