The Hey family of bHLH transcription factors are direct targets of Notch signaling. HEY2 (gridlock) is an important determinant of mammalian heart development, loss of which in mouse leads to cardiac defects with high postnatal lethality. Mouse Hey1 knockouts have no apparent phenotypic defect but the combined loss of Hey1 and Hey2 results in embryonic death after embryonic day 9.5 (E9.5) with a global lack of vascular remodeling and massive hemorrhage. Similar defects are observed in Jagged1 and Notch1 knockout mice.

Suitable for use in Immunohistochemistry. Other applications not tested.

Optimal dilutions to be determined by the researcher.

1. Dissect tissues. Freeze on dry ice. 2. Cut on a cryostat - 10um slices. 3. Dry slides at RT for 30-90 min. 4. Fix with cold Acetone:Methanol (50:50) for 2 min. Either process further or store at -20°C. Alternative is fixation with 4% PFA and treatment with a 0.05% trypsin solution. Incubate for 10 min. at 37°C. Followed by 3 washes with PBS. 5. Let air dry. 6. Treat with PBS for 5 min. 7. Incubate in 50mM ammonium chloride for 30 min. 8. Treat with PBS for 5 min. 9. Treat with Blocking serum for 30-45 min. 10. Primary antibody, H2100-10, for 90 min. Dilution: 1:100-1:600 11. Wash 3 times for 5 min. 12. Secondary fluorescent conjugated antibody for 30 min. 13. Wash 3 times for 5 min. and mount.

Store product at 4°C in the dark. DO NOT FREEZE! Stable at 4°C for 12 months after receipt as an undiluted liquid. Dilute required amount only prior to immediate use. Further dilutions can be made in assay buffer. Caution: MaxLight™550 conjugates are sensitive to light. For maximum recovery of product, centrifuge the original vial prior to removing the cap.

Note: Applications are based on unconjugated antibody.

Synthetic peptide corresponding to aa314-330 from human Hey 2/HRT2.

Supplied as a liquid in PBS, pH 7.2. No preservative added. Labeled with MaxLight™550.

Purified by affinity chromatography.

Recognizes human Hey 2/HRT2. Species Crossreactivity: rodent.