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mAbs Against HUMAN Protein

HP-1α

Antibody Validated
CONTACT INFORMATION:Monoclonal Antibodies Unit, Centro Nacional de Investigaciones Oncológicas
STATUS:Validated
TYPE:rat anti human
CLONE NAME:HP330A
PROTEIN:human full length HP1α
PROTEIN WEB:http://www.hprd.org/summary?hprd_id=05131&isoform_id=05131_3&isoform_name=Isoform_1
ANTIGEN USED:GST-HP1α and GST-HP1γ recombinant proteins
FUSION PARTNER:NS-1
ISOTYPE:IgG2a
SPECIES REACTIVITY:Human
PREPARATION AND STORAGE:Aliquot and store at 4C. Do not freeze
APP RECOMMENDED:IHQ-paraffin, IHQ-frozen, IF, WB
APP NO TESTED:IP, Flow cytometry

DESCRIPTION

Heterochromatin protein-1 (HP1) is a methyl-lysine binding protein localized at heterochromatin sites, where it mediates gene
silencing. HP-1alpha is involved inthe formation of functional kinetochore through interactionwith MIS12 complex proteins. HP-
1alpha also interacts with lamin B receptor, contributing to the association of the heterochromatin with the inner nuclear membrane.

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
HP330A 1:20 supernatant None rabbit anti rat HRP Tonsil / Nuclear not done
Paraffin tissue
Recommended Result obtained is satisfactory. The reagent can be use in this application
HP330A 1:20 upernatant 20 min ER2 Tonsil / Nuclear not done Available
Immunofluorescence
Recommended Result obtained is satisfactory. The reagent can be use in this application
HP330A neat upernatant 20 min ER2 Tonsil / Nuclear not done Available
Enlarge image
HP1 (330A) IHC in transfected cytospins and by ELISA
Heterochromatin protein-1 (HP1) is a methyl-lysine binding protein localized at heterochromatin sites and expressed in all cell types. We made a mAb against HP1 protein using, as immunogen, a combination of HP1 alpha and HP1 gamma protein. We could not distinguish by IHC in tranfected Hek HP1 alpha and HP1 gamma which subunit was recognized by our antibody since strong nuclear staining was observed in both cytospins. Using ELISA technique we could determine that our antibody is specific for the alpha subunit.
Enlarge image
Rat monoclonal 330A can be used to detect HP1 α expression in frozen human tissues.
Enlarge image
Rat monoclonal 330A can be used to detect HP1 α expression in paraffin human tissue
Enlarge image
Immunofluorescence of HP1 (330A) mAb in Ramos cell line
WB Techniques
WB Techniques Clone Dilution Antibody concentration +/- control Expected MW Observed Mw Positivity in other species Protocol
Western Blotting
Recommended Result obtained is satisfactory. The reagent can be use in this application
HP330A neat supernatant Hela / 22kDa 22kDa Not done Available
Immunoprecipitation
Enlarge image
Rat monoclonal HP330A can be used to detect HP1α protein in human cell lines by WB.
Lanes
Lane 1 Transfected Hek-V5-HP1 α (10цg) (+)
Lane 2 Transfected Hek-V5-HP-1 γ (10цg) (-)
Lane 3 Transfected Hek-V5-P16 (10цg) (-)
Lane 4 Hela cell line (100цg) (+)
Lane 5 Jurkat cell line (100цg) (+)
Lane 6 Ramos cell line (100 цg) (+)

Rabbit antibody from Cell Signaling was used at 1:800