HEY 2 (Helix-loop-helix Transcription Factor, Hairy/enhancer-of-split related with YRPW motif 2, bHLHb32, BHLHB32, Cardiovascular helix-loop-helix factor 1, CHF1, Class B basic helix-loop-helix protein 32, GRIDLOCK, GRL, Hairy and enhancer of split-relate

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.\n\nApplications: \nSuitable for use in Immunohistochemistry. Other applications not tested.\n\nRecommended Dilutions:\nImmunohistochemistry: 1:200-1:1000\nOptimal dilutions to be determined by the researcher.\n\nImmunohistochemistry Protocol:\n1. Dissect tissues. Freeze on dry ice.\n2. Cut on a cryostat - 10um slices.\n3. Dry slides at RT for 30-90 min.\n4. Fix with cold Acetone:Methanol (50:50) for 2 min. Either process further or store at -20 degrees C. Alternative is fixation with 4% PFA and treatment with a 0.05% trypsin solution. Incubate for 10 min. at 37 degrees C. Followed by 3 washes with PBS.\n5. Let air dry.\n6. Treat with PBS for 5 min.\n7. Incubate in 50mM ammonium chloride for 30 min.\n8. Treat with PBS for 5 min.\n9. Treat with Blocking serum for 30-45 min.\n10. Primary antibody, H2100-10, for 90 min. Dilution: 1:100-1:600\n11. Wash 3 times for 5 min.\n12. Secondary fluorescent conjugated antibody for 30 min.\n13. Wash 3 times for 5 min. and mount.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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