Technical Data
I0650-01A
ICOS (CD278, Inducible T cell co stimulator, Activation inducible lymphocyte immunomediatory molecule, AILIM, MGC39850) (Biotin)
Description:
The inducible costimulator (ICOS, T cell activation molecule H4) is similar to human CD28 (24% homology), and plays an analogous role in the T cell activation process. Secondary signaling through CD28 or ICOS results in discrete cytokine secretion profiles by the activated T cells(1). Signaling by either molecule is effectively down regulated by CD152 (CTLA-4) engagement(2). Human GL50 is a member of the B7 family sharing ~20% homology with CD80 (B7-1) and CD86 (B7-2), and has been shown to be the ligand for ICOS(3). Two RNA splice variants exist for this molecule, differing only in the cytoplasmic domain.

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

Recommended Dilution:
Functional Application: Blocks binding of recombinant GL50-muIg to HPB-MLT cells.
Flow cytometry: 1ug/ml
Optimal dilutions to be determined by the researcher.

Protocol:
1. Five x 10e5 cultured human HPB-MLT cells were washed and incubated 45 minutes on ice with 80ul of I0650-01A at 1ug/ml.
2. Cells were washed twice and incubated with Streptavidin(PE) (S7973-88P) after which they were washed three times, fixed and analyzed by FACS. Cells stained positive with a mean shift of 1.38 log10 fluorescent units when compared to a Mouse IgG1(Biotin) negative control at a similar concentration.
3. Binding was blocked when cells were pre incubated 10 minutes with 20ul of 0.5mg/ml anti-CD278 antibody (I0650-01).

Storage and Stability:
May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
TypeIsotypeCloneGrade
MabIgG1,k7H152Affinity Purified
SizeStorageShippingSourceHost
100ug-20CBlue IceHumanMouse
Concentration:
~0.5mg/ml
Immunogen:
Human HPB-MLT cells, recombinant human ICOS-muIg.
Purity:
Purified by Protein A affinity chromatography.
Form
Supplied as a liquid in PBS, pH 7.5, 0.2% BSA, 0.04% sodium azide, 5% glycerol. Labeled with Biotin.
Specificity:
Recognizes human ICOS. Binds to recombinant ICOS-muIg in ELISA, and to the cell surface of HPB-MLT cells in flow cytometry.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1) Beier, K.C., R.A. Kroczek, et al. 2000, Eur J Immunol. 30(12):3707-3717.
2) Riley, J.L., C.H. June, et al. 2001, J. Immunol.166 : 4943-4948.
3) Ling, V., M. Collins, et al. 2000, J. Immunol.164: 1653-1657.
4) Ling, V., M. Collins, et al. 2001, J. Immunol.166: 7300-7308.