Working principle of the desalting column: It distinguishes different substances according to their molecular sizes. Due to the obvious difference in size between salt molecules and the target molecules in the sample (such as proteins), the separation of target molecules from salt molecules can be achieved.
Schematic diagram of the operation process
Product Components and Storage Conditions:
Item Number
| Component
| 96T
| Storage
|
P1456352A
| Peptide Desalting Columns
| 96T
| RT
|
H1456342A
| Wash buffer 1
| 35 mL
| RT
|
H1456342B
| Wash buffer 2
| 24 mL
| RT
|
H1456342C
| Wash buffer 3
| 24 mL
| RT
|
H1456342D
| Elution buffer
| 50 mL
| RT. Store in the dark.
|
H1456342E
| Loading buffer
| 2 mL
| RT
|
P1456352B
| Collection Tube
| 30个
| 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:
1.During the sample loading process, the packing in the desalting column must be kept moist.
2.If the sample contains a high concentration of salt (such as 2M urea or >100nM sodium bicarbonate), repeat the rinsing step 1-2 times.
3.This product is for one-time use only; multiple uses are not recommended.
4.This product is limited to scientific research use by professional personnel, and must not be used for clinical diagnosis or treatment, nor for food or drugs.
