TMB 1-Component HRP Microwell Substrate (SUBT)

Biorbyt

TMB 1-Component HRP Microwell Substrate (SUBT)

10

Flow Cytometry, 1. Each 50 microL vial is sufficient for 1000 flow cytometry tests., 2. Spin thawed vial briefly in a microcentrifuge to remove any reagent that may have become trapped in the cap., 3. Dilute Green Live/Dead Stain concentrated stock solution (500 microM) 1:100 in PBS to prepare a 5000 nM working solution. For example, add 10 microL stock concentrate to 990 microL PBS. Prepare only what is needed for the experiment. Working solution should be used immediately, and any remaining diluted solution should be discarded., 4. Spike samples with a 1:100 dilution of the 5000 nM working solution. For example, spike 0.495 mL samples with 5 microL working solution., 5. Incubate samples for ~10 minutes at room temperature, protected from light., 6. Analyze with a flow cytometer using a blue laser at 488 nm and a 530/30 (FL1) emission filter setting, or similar., Fluorescence Microscopy, 1. Each 50 microL vial is sufficient for 100 microtiter plate wells., 2. Spin thawed vial briefly in a microcentrifuge to remove any reagent that may have become trapped in the cap., 3. Dilute Green Live/Dead Stain concentrated stock solution (500 microM) 1:100 in PBS to prepare a 5000 nM working solution. For example, add 10 microL stock concentrate to 990 microL PBS. Prepare only what is needed for the experiment. Working solution should be used immediately, and any remaining diluted solution should be discarded., 4. Spike samples with a 1:10 dilution of the 5000 nM working solution. For example, spike .45 mL samples with 50 microL working solution., 5. Incubate samples for ~10 minutes at room temperature, protected from light., 6. When bound to DNA, the peak absorption of Green Live/Dead Stain is 495 nm and the maximum emission is 512 nm. Visualize with a fluorescence microscope using optical filters that best approximate these settings.

Research Use Only

A ready-to-use solution containing 3,3,5,5-tetramethylbenzidine in a mildly acidic buffer without aprotic solvents. Ideal for use in all ELISAs where the target detection level is in the ng-pg/mL range and the detection enzyme is HRP-conjugated.

2°C to 8°C

41116133