In Vivo (Add solvents to the product individually and in order.)
Homogeneous suspension
CMC-NA
≥5mg/ml
Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution
5%DMSO
40%PEG300
5%Tween80
50%ddH2O
Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.
0.100mg/ml
(0.25mM)
Taking the 1 mL working solution as an example, add 50 μL of 2 mg/ml clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to clarify; then continue to add 500 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.
Clear solution
5% DMSO
95% Corn oil
Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.
0.100mg/ml
(0.25mM)
Taking the 1 mL working solution as an example, add 50 μL of 2 mg/ml clear DMSO stock solution to 950 μL of corn oil and mix evenly. The mixed solution should be used immediately for optimal results.
* <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
BAM 7 is a direct and selective activator of proapoptotic Bax with an EC50 of 3.3
M.
Targets
Bax
3.3 μM
In vitro
BAM7 directly binds the previously uncharacterised BH3-binding groove at the N-terminal face of BAX. This compound is selective for the BH3-binding groove at the N-terminal face of BAX. It directly interacts with BAX at the very surface used by the BIM BH3 helix to trigger BAX activation. This chemical results in functional BAX activation. It triggers the conversion of BAX from monomer to oligomer in a dose- and time-responsive manner, the kinetics of which approach saturation at a 1:8 dose ratio of BAX:BAM7. This compound triggers in vitro BAX oligomerisation, BAX-mediated pore formation and BAX-dependent cell death. It selectively induces BAX-mediated Apoptosis by triggering the hallmark features of intracellular BAX activation. This chemical only kills the cell line that contains BAX, eliciting the biochemical and morphologic features of BAX-mediated Apoptosis.
Features
Does not interact with the BH3-binding pocket of antiapoptotic proteins or proapoptotic BAK and induces cell death in a BAX-dependent fashion.
Direct binding curves are first generated by incubating FITC-BIM SAHB (50 nM) with serial dilutions of full-length BAX, BCL-XLΔC, MCL-1ΔNΔC, BFL-1/A1ΔC or BAKΔC and fluorescence polarisation measured at 20 minutes on a SpectraMax M5 microplate reader. For competition assays, a serial dilution of this compound or acetylated BIM SAHB (Ac-BIM SAHB) is combined with FITC-BIM SAHB (50 nM), followed by the addition of recombinant protein at ~EC75 concentration, as determined by the direct binding assay (BAX, BAKΔC: 500 nM; BCL-XLΔC, MCL-1ΔNΔC, BFL-1/A1ΔC: 200 nM). Fluorescence polarisation is measured at 20 minutes and IC50 values calculated by nonlinear regression analysis of competitive binding curves using Prism software.
MEFs (2.5 × 103 cells per well) are seeded in 96-well opaque plates for 18-24 h and then incubated with serial dilutions of BAM7, ANA-BAM16 or vehicle (0.15% (v/v) DMSO) in DMEM at 37 °C in a final volume of 100 μL. Cell viability is assayed at 24 h by addition of CellTiter-Glo reagent according to the manufacturer’s protocol, and luminescence is measured using a SpectraMax M5 microplate reader. Viability assays are performed in at least triplicate, and the data are normalized to vehicle-treated control wells.
References
https://pubmed.ncbi.nlm.nih.gov/22634637/
Sellecks BAM7 Has Been Cited by 3 Publications
Small-molecule allosteric inhibitors of BAX.
[ Nat Chem Biol, 2019, 15(4):322-330]
Drug manipulation of pro-and anti-apoptotic Bcl-2 proteins in the regulation of Parkin-mediated mitophagy in human breast cancer cells
[ Baltimore, Maryland , 2018, ]
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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.
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