HES4 (Hairy Family Helix-loop-helix Transcription Factor)

HES-4 is a 221 aa, 24 kD member of the HES-family of transcriptional repressors. HES-4 contains a species-specific bHLH domain (aa 33-93), a specific orange DNA-binding domain (aa 109-150), a 65 aa Proline-rich SH3-binding domain, and a WRPW Groucho/TLE interacting C- terminal motif. HES-4 is a HES-1 paralog likely to maintain stem cell pluripotency. In the region used as immunogen, the amino acid sequence of human and canine HES-4 are 80% identical.\n\nApplications: \nWestern Blot: This antibody can be used at 0.1-0.2ug/ml with the appropriate secondary reagents to detect human HES-4. The detection limit for rhHES-4 is approximately 5 ng/lane under non-reducing and reducing conditions. \nImmunocytochemistry: This antibody has been used at a concentration of 10ug/ml to detect HES-4 in HEK293 cells. Cells were fixed with PBS containing 4% paraformaldehyde for 20 minutes at room temperature and blocked with PBS containing 10% normal donkey serum, 0.1% Triton X-100, and 1% BSA for 45 minutes at room temperature. After blocking, cells were incubated with diluted primary antibody overnight at 4 degrees C followed by fluorochrome-coupled anti-sheep IgG at room temperature in the dark for one hour. Between each step, cells were washed with PBS containing 0.1% BSA. \nDirect ELISA: This antibody can be used at 0.5-1.0ug/ml with the appropriate secondary reagents to detect human HES-4. The detection limit for rhHES-4 is approximately 0.2 ng/well.\nOptimal dilutions should be determined by each laboratory for each application.\n\nStorage and Stability:\nLyophilized powder may be stored at 4 degrees C for short-term only. Reconstitute to nominal volume by adding sterile PBS with 40-50% glycerol and store at -20 degrees C. Reconstituted product is stable for 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

H2034-38A

Size

100ug

Applications

ELISA, ICC, WB

Hosts

Sheep

Reactivities

Hum

Form

Supplied as a lyophilized powder from a 0.2um filtered solution in PBS with 5% trehalose. Reconstitute with sterile PBS, 40-50% glycerol. If 0.5ml is used, the concentration will be 0.2mg/ml.

P Type

Pab

Purity

Purified by human HES-4 affinity chromatography.

Isotype

IgG

Additional Info

Selected for its ability to recognize human HES-4 in the applications listed below. In direct ELISAs and Western Blots, this antibody shows less than 1% crossreactivity with rhHES-1, rhHES-2, rhHES-5, and rhHES-7.