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Applications
Signature Peptides of Rituximab on a bioZen Peptide XB-C18 Column Using bioZen MagBeads
25617
Separation Mode: Reversed Phase
Reversed Phase
PEPTIDE XB-C18
HPLC
Pharmaceutical/Biopharmaceutical
Therapeutic / Clinical
Biochemical Compound / Nutrient
Applications
Signature Peptides of Rituximab on a bioZen Peptide XB-C18 Column Using bioZen MagBeads
Analytes
Details
LC Conditions (App ID: 25617)
Column: | ||||||||||||
Brand Name: | ||||||||||||
Part No: | ||||||||||||
Phase Name: PEPTIDE XB-C18 | ||||||||||||
Elution Type: Gradient | ||||||||||||
Mobile Phase: A: 0.1% Formic acid in Water B: 0.1% Formic acid in Acetonitrile | ||||||||||||
Gradient Profile:
| ||||||||||||
Flow Rate: 0.3 mL/min | ||||||||||||
Temperature: 40°C | ||||||||||||
System: Agilent Technologies Agilent Technologies | ||||||||||||
Detection: LC/MS | ||||||||||||
Sample Note: *Rat |
Sample Preparation Details
Description: bioZen MagBeads, Streptavidin Coated - 500mg, Ea |
Part Number: |
Evaporation: Evaporate to dryness |
Reconstitute: beads with PBS buffer to the original slurry volume. |
Sample Notes: *Rat |
0 Dispense 25 µL of bead slurry into a 96 well plate |
1 Transfer 96 well plate onto magnetic stand for 30 seconds and discard liquid |
2 Transfer 96 well plate onto magnetic stand for 30 seconds and discard liquid |
3 Vortex bead slurry using mixer or pipette |
4 Transfer 96 well plate onto magnetic stand for 30 seconds and discard liquid |
5 Add 500µL PBS Buffer to each plate well that contains beads |
6 Transfer 96 well plate onto magnetic stand for 30 seconds and discard liquid |
7 Transfer 96 well plate onto magnetic stand for 30 seconds and discard liquid |
8 Transfer 96 well plate onto magnetic stand for 30 seconds and discard liquid |
9 Vortex bead slurry using mixer or pipette |
10 Transfer 50 µL of bead slurry into each well that contains 25 µL sample. Mix well before transferring |
11 Reconstitute beads with PBS buffer to the original slurry volume. |
12 Transfer 96 well plate onto magnetic stand for 30 seconds and discard liquid |
13 Transfer plate for incubation for one hour at room temperature with a shaking speed of 1200 RPM using a deep well plate thermoshaker. Beads should stay suspended during incubation |
14 Transfer 96 well plate onto magnetic stand for 30 seconds and discard liquid |
15 Add 500µL PBS Buffer to each plate well that contains beads |
16 Transfer 96 well plate onto magnetic stand for 5 minutes and collect supernatant |
17 Transfer 96 well plate onto magnetic stand for 30 seconds and discard liquid |
18 Add 500µL PBS Buffer to each plate well that contains beads |
19 Shake and mix the bead slurry, using vortex or pipette |
20 Transfer 96 well plate onto magnetic stand for 30 seconds and discard liquid |
21 Add 500µL PBS Buffer to each plate well that contains beads |
22 Shake and mix the bead slurry, using vortex or pipette |
23 Transfer 96 well plate onto magnetic stand for 30 seconds and discard liquid |
24 Reconstitute beads with PBS buffer to the original slurry volume. |
25 Shake and mix slurry of anti-human IgG coated beads well, using vortex or pipette |
26 Transfer 50 µL of bead slurry into each well that contains 25 µL sample. Mix well before transferring |
27 Add cover to plate using polyester plate film |
28 Vortex to mix plate |
29 Transfer plate for incubation for one hour at room temperature with a shaking speed of 1200 RPM using a deep well plate thermoshaker. Beads should stay suspended during incubation |
30 Transfer 96 well plate onto magnetic stand for 30 seconds and discard liquid |
31 Add 200 µL of PBS Buffer to each well that contains beads |
32 Vortex well and place plate on centrifuge for 2 minutes at 400RPM |
33 Transfer 96 well plate onto magnetic stand for 30 seconds and discard liquid |
34 Add 200µL of 10mM Ammonium Bicarbonate to each well that contains beads |
35 Vortex well and place plate on centrifuge for 2 minutes at 400RPM |
36 Transfer 96 well plate onto magnetic stand for 30 seconds and discard liquid |
37 Add 100µL of 0.1% TFA in water to each well and vortex |
38 Add pH paper to check pH value (should be lower than 3) |
39 Add plate cover and shake for 10 minutes at 1200RPM using thermomixer |
40 Transfer plate on centrifuge for 2 minutes at 400RPM |
41 Transfer 96 well plate onto magnetic stand for 5 minutes and collect supernatant |
Order Items Used in This Application
00B-4768-AN
bioZen 2.6 µm Peptide XB-C18, LC Column 50 x 2.1 mm, Ea
Qty:
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