		Technical Data
B5001-61K	BTLA (B- and T-lymphocyte Associated Protein, B- and T-lymphocyte Attenuator, BTLA1, CD272, CD272 Antigen, FLJ16065, MGC129743) (FITC)

Description:
Human CD272 (BTLA, B and T Lymphocyte Attenuator) is a member of the Immunoglobulin superfamily and has sequence homology to PD-1 and CTLA4. It is expressed on T and B lymphocytes and other hemopoetic lineages. Engagement of this molecule by its ligand, CD270 (HVEM, a TNF superfamily member) can down regulate activated T and B cell responses. CD272 levels on antigen specific CD8+ T cells have been reported to decrease in viral specific, but not melanoma specific activated lines. Polymorphism of the CD272 molecule has been linked to rheumatoid arthritis.

Applications:
Suitable for use in ELISA and Flow Cytometry. Other applications not tested.

Recommended Dilutions:
ELISA: Detection
Flow Cytometry: 1:50
Optimal dilutions to be determined by the researcher.

Flow Cytometry Protocol:
1. 5x10e5 ficoll prepared human peripheral blood mononuclear cells (PBMC) per tube were pelleted and pre incubated 5 minutes with 20ul of human IgG at 300ug/ml, after which they were washed and incubated 45 minutes on ice with 80ul of B5001-61K at a 1:50 dilution (5ug/ml).
2. They were then washed three times, fixed and analyzed by FACS using a lymphoid gate.
3. Cells stained positive with a mean shift of 0.54 log10 fluorescent units when compared to a Mouse IgG1 (FITC) negative control at a similar concentration.
4. Binding was blocked when cells were pre incubated 10 minutes with 20ul of 0.5mg/ml of B5001-61G.

Storage and Stability:
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. FITC conjugates are sensitive to light.

Type	Isotype	Clone	Grade
Mab	IgG1,k	11K3	Affinity Purified

Size		Storage	Shipping	Source	Host
120 tests		-20°C	Blue Ice	Human	Mouse

Concentration:

~0.25mg/ml

Immunogen:

Recombinant protein corresponding to soluble human CD272

Purity:

Purified by Protein A affinity chromatography.

Form

Supplied as a liquid in 50mM sodium phosphate, pH 7.5, 100mM potassium chloride, 150mM sodium chloride, 0.2% BSA, 0.04% sodium azide, 5% glycerol. Labeled with fluorescein isothiocyanate (FITC).

Specificity:

Recognizes human CD272 (BTLA). Binds to CD272 on the cell surface of human PBMC in Flow Cytometry and to recombinant CD272-muIg in ELISA. Does not block binding of Biotinylated CD270 (HVEM)-muIg to CD272-muIg in ELISA, but appears to enhance this interaction.

Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.

1. Derre’ L, DE Speiser, et al. (2010) J Clin Invest 120(1):157-167, 2. Otsuki N, Azuma M, et al. (2006) Biochem Biophys Res Com 344(4): 1121-1127, 3. Lin SC, Chan CH, et al. (2006) J Biomed Sci 13(6): 853-860, 4. Pasero C, D Olive, et al. (2009) Curr Mol Med 9(7): 911-927, 5. Gavrieli M, KM Murphy, et al. (2006) Adv Immunol 92: 152-185.