iHAP1

Catalog No.S9695 Batch:S969501

Technical Data

Formula	C₂₀H₁₄ClNO₂S
Molecular Weight	367.85	CAS No.	105925-39-1
Solubility (25°C)*	In vitro	DMSO	20 mg/mL (54.36 mM)
		Ethanol	5 mg/mL (13.59 mM)
		Water	Insoluble
* <1 mg/ml means slightly soluble or insoluble. * Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations. * Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

Preparing Stock Solutions

Biological Activity

Description

iHAP1 is a well-established microtubule poison, blocks microtubule polymerization fully at 5 µM which is independent of deregulated transcription, causes marked sensitization when deleting Rod1 gene in HeLa cells.

Targets

microtubule polymerization ^[1]

In vitro

The depletion of Rod1 and TACC3 results in sensitization to iHAP1, which is caused by chemogenic interaction between Rod1 and iHAP1. As for the RPE1‐hTERT P53−/− background, deletion of Rod1 in HeLa cells causes marked sensitization to iHAP1.^[1]

Protocol (from reference)

Cell Assay:^{^[1]}	Cell lines RPE1‐hTERT P53−/− Flag‐Cas9 cell line Concentrations 0.16-1 µM Incubation Time 12 days Method The Sulforhodamine B (SRB) proliferation assay was used to assess in vitro cytotoxicity of the compounds. Cells were treated with sublethal drug concentrations (LD20) for 12 days from the day of seeding. If necessary, transient siRNA transfection was performed 3 days before drug treatment. After 12 days, medium was removed from the wells and cells were washed once with PBS and fixed in prechilled 10% trichloroacetic acid (TCA) for 30 min at 4°C. Cells were subsequently washed two times with double‐distilled water and stained with 0.4% SRB (1% acetic acid for 20 min at room temperature protected from light). SRB was then removed, and cells were washed four times in 1% acetic acid. After complete light‐protected drying, SRB was dissolved by addition of 10 mM Tris pH 8 and gentle shaking at room temperature for 2 h. The absorbance at A510 was read. Percentage of cell growth inhibition was calculated. The IncuCyte® live‐cell analysis system was additionally used to real‐time track the growth of the cells subjected to the different treatments.

Cell Assay:^{^[1]}

Cell lines
RPE1‐hTERT P53−/− Flag‐Cas9 cell line
Concentrations
0.16-1 µM
Incubation Time
12 days
Method
The Sulforhodamine B (SRB) proliferation assay was used to assess in vitro cytotoxicity of the compounds. Cells were treated with sublethal drug concentrations (LD20) for 12 days from the day of seeding. If necessary, transient siRNA transfection was performed 3 days before drug treatment. After 12 days, medium was removed from the wells and cells were washed once with PBS and fixed in prechilled 10% trichloroacetic acid (TCA) for 30 min at 4°C. Cells were subsequently washed two times with double‐distilled water and stained with 0.4% SRB (1% acetic acid for 20 min at room temperature protected from light). SRB was then removed, and cells were washed four times in 1% acetic acid. After complete light‐protected drying, SRB was dissolved by addition of 10 mM Tris pH 8 and gentle shaking at room temperature for 2 h. The absorbance at A510 was read. Percentage of cell growth inhibition was calculated. The IncuCyte® live‐cell analysis system was additionally used to real‐time track the growth of the cells subjected to the different treatments.

References

[1] Vit G, et al. EMBO J. 2022 Jul 18;41(14):e110611.

Selleck's iHAP1 has been cited by 1 publication

Non-catalytic allostery in α-TAT1 by a phospho-switch drives dynamic microtubule acetylation [ J Cell Biol, 2022, 221-11e202202100]	PubMed: 36222836

Non-catalytic allostery in α-TAT1 by a phospho-switch drives dynamic microtubule acetylation [ J Cell Biol, 2022, 221-11e202202100]

PubMed: 36222836

RETURN POLICY
Selleck Chemical’s Unconditional Return Policy ensures a smooth online shopping experience for our customers. If you are in any way unsatisfied with your purchase, you may return any item(s) within 7 days of receiving it. In the event of product quality issues, either protocol related or product related problems, you may return any item(s) within 365 days from the original purchase date. Please follow the instructions below when returning products.

SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

NOT FOR HUMAN, VETERINARY DIAGNOSTIC OR THERAPEUTIC USE.