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.
Applications:Western 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.
Immunocytochemistry: 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° 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.
Direct 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
Optimal dilutions should be determined by each laboratory for each application.
Storage and Stability:Lyophilized powder may be stored at 4°C for short-term only.
Reconstitute to nominal volume by adding sterile PBS with 40-50% glycerol and store at -20°C.
Reconstituted product is stable for 12 months at -20°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.
仕様
Size：100ug
Host：sheep
Source Antibody：human
Grade：Affinity Purified
Purity：Purified by human HES-4 affinity chromatography.
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.
Specificity：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.
