Supporting Reagent with the desalting column can handle the desalting step after enzymatic hydrolysis, quickly prepare pretreated samples, and are used for subsequent mass spectrometry analysis.
Schematic diagram of the operation process
Product Components and Storage Conditions:
H1456342
| Component
| 25T
| 48T
| 96T
| Storage
|
H1456342A
| Wash buffer 1
| 9.5 mL
| 17.5 mL
| 35 mL
| RT
|
H1456342B
| Wash buffer 2
| 6 mL
| 12 mL
| 24 mL
| RT
|
H1456342C
| Wash buffer 3
| 6 mL
| 12 mL
| 24 mL
| RT
|
H1456342D
| Elution buffer
| 12 mL
| 25 mL
| 50 mL
| RT. Store in the dark.
|
H1456342E
| Loading buffer
| 500 μL
| 1 mL
| 2 mL
| RT |
Operating Procedure:
① Add 320μL of Wash buffer 1 to the sample after enzymatic hydrolysis, shake vigorously for 3 minutes, centrifuge at 15000rpm for 3 minutes, and remove the upper liquid.
② Transfer the lower layer sample into the Tip column, centrifuge at 2500rpm for 3-5 minutes until all the liquid is centrifuged down. If the liquid flow rate is slow, the rotation speed can be increased.
③ Add 200μL of Wash buffer 2 (shake for 10-20 seconds before use) to the desalting column, centrifuge at 2500rpm for 3-5 minutes until all the liquid is centrifuged down.
④ Add 200μL of Wash buffer 3 to the desalting column, centrifuge at 2500rpm for 3-5 minutes until all the liquid is centrifuged down.
⑤ Put the desalting column into a new EP tube, add 200μL of Elution buffer to the desalting column, centrifuge at 2000rpm for 3-5 minutes until all the liquid is centrifuged down.
⑥ Repeat step ⑤, collect the eluates from both times, and freeze-dry them.
⑦ Add 10μL of Loading buffer, vortex vigorously for 3 minutes, centrifuge at 20000g for 10 minutes, take an appropriate amount of sample for mass spectrometry detection. Taking the HF-X instrument as an example, 0.5-1μg of sample is sufficient for loading.
Precautions and Disclaimer:
This product is limited to scientific research use by professional personnel. It must not be used for clinical diagnosis or treatment, nor for food or drugs.
ZJ25F0927267