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mAbs Against HUMAN Protein

BTLA

Antibody Validated
CONTACT INFORMATION:Monoclonal Antibodies Unit, Centro Nacional de Investigaciones Oncológicas
STATUS:Validated
TYPE:mouse anti human
CLONE NAME:FLO67B
PROTEIN:human full length BTLA protein
PROTEIN WEB:http://www.ncbi.nlm.nih.gov/protein/145580621
ANTIGEN USED:HIS-BTLA
FUSION PARTNER:NS1/Ag4-1 (NS1) cells
ISOTYPE:IgG1
SPECIES REACTIVITY:human
PREPARATION AND STORAGE:Aliquot and store at 4C. Do not freeze
APP RECOMMENDED:IHQ-frozen, IHQ-paraffin, IF
APP NO RECOMMENDED:WB
APP NO TESTED:IP, Flow cytometry

DESCRIPTION

B and T lymphocyte attenuator (BTLA) has recently been identified as a new inhibitory receptor of the CD28 superfamily, with similarities to cytotoxic T lymphocyte activation antigen (CTLA)-4 and programmed death (PD)-1. Engagement of BTLA on T lymphocytes can profoundly reduce the T cell receptor (TCR)-mediated activation.

APPLICATIONS

IHC Techniques
IHC Techniques Clone Dilution Antibody concentration Antigen retrieval method Visualization kit +/- control Protein localization Positivity in other species Protocol
Frozen tissue and cytospins
Recommended Result obtained is satisfactory. The reagent can be use in this application
FLO67B 1:5 supernatant Tonsil / Cytoplasmic
Paraffin tissue
Recommended Result obtained is satisfactory. The reagent can be use in this application
FLO67B 1:50 supernatant 20min ER2 (Tris-EDTA) Novolink kit Tonsil / Kidney Cytoplasmic Available
Immunofluorescence
Recommended Result obtained is satisfactory. The reagent can be use in this application
FLO67B neat supernatant Tonsil / Citoplasmic Available
Enlarge image
BTLA antibody (FLO67B) in transfected cells
Cytoplasmic staining of BTLA mAb in transfected HEK-V5-BTLA cells. Anti-V5 mAb was used as positive control. HEK-V5-B-MYB transfected cells were used as a negative control.
Enlarge image
BTLA expression on human frozen tonsil.
Enlarge image
FLO67B staining in human paraffin tissues.
Enlarge image
BTLA (FLO67B) immunofluorescence in human paraffin tonsil.
Anti-BTLA (FLO67B) is represented in green. anti-PD1 is represented in red. DAPI in blue.
WB Techniques
WB Techniques Clone Dilution Antibody concentration +/- control Expected MW Observed Mw Positivity in other species Protocol
Western Blotting
Not recommended Reagent discarded for this application
FLO67B neat supernatant / Available
Immunoprecipitation
Enlarge image
Anti-BTLA (FLO67B) antibody in Western Blotting
FLO67B antibody is not working well by WB. The MW observed is around 35kDa. The antibody is not validated for WB since the results obtained are not reproducible at the moment.

Lane 1 Hek-BTLA (30ug)
Lane 2 Hek-Bcl7b (30ug)
Lane 3 Tonsil (100ug)
Lane 4 DOHH2 cell line (100ug)
Lane 5 Daudi cell line (100ug)
Lane 6 YT cell line (100ug)
Lane 7 Karpas 422 cell line (100ug)
Lane 8 REH cell line (100ug)
Lane 9 LP1 cell line (100ug)