TMB 1-Component HRP Microwell Substrate (SUBT)

SKU: orb699915

Description

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.

Images & Validation

• Application Notes:
  Flow Cytometry, 1. Each 50 µL 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 µM) 1:100 in PBS to prepare a 5000 nM working solution. For example, add 10 µL stock concentrate to 990 µL 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 µL 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 µL 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 µM) 1:100 in PBS to prepare a 5000 nM working solution. For example, add 10 µL stock concentrate to 990 µL 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 µL 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.

Storage & Handling

• Storage: 2-8°C
• Concentration: 1 X
• Disclaimer: For research use only