Inhibitor Handling
Instructions

Storage

3 years -20°C powder
As a powder, the inhibitor can be stored at-20°C for up to 3 years.

Aliquot the stock solution to routine usage volumes and store at -20°C or -80°C. Avoid
repeated freezing and thawing.

5. How to prepare the working solution?
1. How to deal with the product(s) after receiving?
The packaging of the product(s) may have turned upside down during transportation, resulting
in the inhibitor adhering to the neck or cap of the vial. Take the vial out of its packaging andgently shake to let the inhibitor fall to the bottom of the vial. For liquid products, centrifuge at
200-500 RPM to gather the liquid at the bottom of the vial. Try to avoid loss or contamination
during handling.

2. What is the range of the weighing errors for ourproducts?
Weighing range

Error range

5 - 25 mg
50 -500 mg>1 g

0.1 mg
1 mg
2 - 5 mg

(1) Calculate the dilution required by using our dilution calculator.
http://www.selleckchem.com/dilutioncalculator.jsp(2) Slowly add the stock solution into the solvent until obtaining the desired concentration.
Mix by vortexing or repeated pipetting.
(3) Add a drop of working solution onto a slide and check whether there is precipitation
under the microscope. If present, allow the working solution to stand for 10 min, vortex or
pipette and then recheck whether there is precipitation under the microscope. If there is
still the precipitation, please send an email to techbio@selleckchem.com for help.
(4) Our compounds are mainly liposoluble, so it is common that there is precipitation when
diluted by aqueous solvent such as cell culture medium or PBS. They can be completelydissolved by ultrasonication.

6. How do I sterilize the working solution?
We suggest you to use filters to sterilize the working solution. DO NOT sterilize the working
solution by autoclaving.

3. How do you prepare stock solutions?

7. What issues need special attention for cell-based
assays?

According to your experimental design, select a suitable solvent to prepare a concentrated
stock solution. The solubility information, tested by our Quality Control Department, can befound on the product page of our website. For in vitro experiments, DMSO is often used as thesolvent. It is then diluted to make a working solution (e.g. 1:1000 dilution in cell culture
medium). A typical table for solubility is outlined below:(1) DMSO is used to prepare the stock solution in most cell-based assays. The stock solution
is then diluted in the culture medium to prepare a working solution. Make sure the concentration of DMSO is < 0.3% to avoid poisoning the cells. A negative control in the experiment isusually the culture medium with DMSO at the same concentration.

Solubility (25°C)(2) Our compounds are mainly liposoluble, so it is common that there is precipitation when
diluted by the cell culture medium. However, they can be completely dissolved byultrasonication. If there is still the precipitation, please send an email to techbio@selleckchem.com for help.

DMSO X mg/mL*
Water slightly or not soluble
Ethanol slightly or not soluble*The solubility value is obtained from our technical team; the product can be easilydissolved in DMSO
at X mg/mL.The solubility value may higher, but our technical team did not measure for a higher value.

If you are having difficulties dissolving an inhibitor, we suggest you:
(1) Recalculate the concentration of the stock solution, according to the following formula:
the actual concentration (mg/mL) = molecular weight (g/mol) × concentration (mM) × 10-3
(2) Check whether the solvent has been contaminated, e.g. DMSO absorbs moisture.
(3) Some inhibitors are difficult to dissolve due to their structure or chemical characteristics
and require additional mixing methods such as vortexing or ultrasonication. The inhibitor
may need to be untrasonicated for 1 h or overnight. If necessary, heat the inhibitors nohigher than 50°C to prevent altering the product.
(4) Send an email to techbio@selleckchem.com and let our tech support staff know if you
cannot dissolve the inhibitor completely. We will try to provide additional assistance.

4. How do I store the inhibitor(s)?
You can find the inhibitor storage recommendations on the product webpage. Here are some
general guidelines:

Toll Free:(877)796-6397---USA and Canada only---

Phone:+1-832-582-8158

8. What issues need special attention for animalexperiments?
(1) Our compounds are mainly liposoluble. If DMSO is used to prepare the stock solution
and then diluted to prepare the working solution for animal experiments, it may not be
possible to obtain the required doses. Instead, hydrotropy agents, such as sodium
carboxymethyl cellulose (CMC-Na), Tween 80, or glycerol, are needed. Please send anemail to techbio@selleckchem.com if you require further assistance.
(2) Administration volumes considered good practice (and possible maximal dose volumes):SpeciesRoute and volumes（mL/kg）
Orals.c.

i.p.

i.m.

i.v. (bolus)

Mouse
Rat
Rabbit10 (50)10 (40)20 (80)0.05 (0.1)

5i.v. (slow inj.)(25)

10 (40)5 (10)

10 (20)0.1 (0.2)

5(20)

10 (15)1 (2)5 (20)

0.25 (0.5)

2(10)

Dog

5 (15)1 (2)

1 (20)

0.25 (0.5)

2.5(5)For more details, please read: Diehl KH, et al. J Appl Toxicol, 2001, 21(1), 15-23.

www.selleckchem.com

info@selleckchem.com

(3) Drugs that are administrated orally sometimes cannot be made into a clear solution due tolimitations in solubility. It may not influence the efficacy of the drugs by gavage using
suspensions or emulsions.
(4) If DMSO is used to prepare the stock solution, make sure the concentration of DMSO is <5% to avoid toxicity.

9. What issues need attention for special products?
(1) For oily compounds, add DMSO to dissolve the compound and prepare a stock solution
with a concentration according to the volume available. You do not need to weigh thecompound.
(2) For unstable compounds, add nitrogen to the vial when preparing stock solutions.

(5) Conversion of different model animals based on BSA
Species

Weight (kg)Body Surface Area (m2)

Km factor

BaboonDog
MonkeyRabbitGuinea pigRat
Hamster
Mouse

1210
3
1.8
0.4
0.15
0.08
0.02

0.6
0.5
0.24
0.15
0.05
0.025
0.02
0.007

20
20
12
12
8
6
5
3

10. What are some safety precautions I must takewhen using products from Selleck Chemicals?
You can download the material safety data sheet (MSDS) from the product page for hazard
identification, first aid measures, fire fighting measures, etc.

Animal A (mg/kg) = Animal B (mg/kg) multiplied by Animal B Km
Animal A Km
Value based on data from FDA Draft Guidelines: Center for Drug Evaluation and Research, Center for
Biologics Evaluation and Research. (2002) Estimating the safe starting dose in clinical trials fortherapeutics in adult healthy volunteers, U.S. Food and Drug Administration, Rockville, Maryland, USAFor example, to convert the dose of resveratrol used in a mouse (22.4 mg/kg) to a dose
based on surface area for rat, multiply 22.4 mg/kg by the Km factor for a mouse and then divideby the Km factor for a rat .This calculation results in a rat equivalent dose for resveratrol of 11.2
mg/kg.
mouse Km(3)Rat dose (mg/kg) = mouse dose (22.4 mg/kg) × rat K (6)= 11.2 mg/kg
mToll Free:(877)796-6397---USA and Canada only---

Phone:+1-832-582-8158

www.selleckchem.com

info@selleckchem.com