Applications

Signature Peptides of Rituximab on a bioZen Peptide XB-C18 Column Using bioZen MagBeads

Analytes

Analytes not found

Details

LC Conditions (App ID: 25617)

Column:

bioZen 2.6 µm Peptide XB-C18, LC Column 50 x 2.1 mm, Ea

Brand Name:

bioZen

Part No:

00B-4768-AN

Elution Type:

Gradient

Mobile Phase:

A: 0.1% Formic acid in Water

B: 0.1% Formic acid in Acetonitrile

Gradient Profile:

Step No.	Time(min)	%A	%B
1	0	85	15
2	4.5	75	25

Flow Rate:

0.3 mL/min

Temperature:

40°C

System:

Agilent Technologies Agilent Technologies

Detection:

LC/MS

Sample Preparation Details

Description: bioZen MagBeads, Streptavidin Coated - 500mg, Ea
Part Number: KS0-9533
Sample Notes:
1 Dispense 25 µL of bead slurry into a 96 well plate
2 Transfer 96 well plate onto magnetic stand for 30 seconds and discard liquid
3 Add 500µL PBS Buffer to each plate well that contains beads
4 Vortex bead slurry using mixer or pipette
5 Transfer 96 well plate onto magnetic stand for 30 seconds and discard liquid
6 Add 500µL PBS Buffer to each plate well that contains beads
7 Vortex bead slurry using mixer or pipette
8 Transfer 96 well plate onto magnetic stand for 30 seconds and discard liquid
9 Add 500µL PBS Buffer to each plate well that contains beads
10 Vortex bead slurry using mixer or pipette
11 Transfer 96 well plate onto magnetic stand for 30 seconds and discard liquid
12 Reconstitute beads with PBS buffer to the original slurry volume.
13 Add 10 µL of biontinylated goat anti-human IgG (0.5mg/mL).
14 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
15 Transfer 96 well plate onto magnetic stand for 30 seconds and discard liquid
16 Add 500µL PBS Buffer to each plate well that contains beads
17 Shake and mix the bead slurry, using vortex or pipette
18 Transfer 96 well plate onto magnetic stand for 30 seconds and discard liquid
19 Add 500µL PBS Buffer to each plate well that contains beads
20 Shake and mix the bead slurry, using vortex or pipette
21 Transfer 96 well plate onto magnetic stand for 30 seconds and discard liquid
22 Add 500µL PBS Buffer to each plate well that contains beads
23 Shake and mix the bead slurry, using vortex or pipette
24 Transfer 96 well plate onto magnetic stand for 30 seconds and discard liquid
25 Reconstitute beads with PBS buffer to the original slurry volume.
26 Shake and mix slurry of anti-human IgG coated beads well, using vortex or pipette
27 Transfer 50 µL of bead slurry into each well that contains 25 µL sample. Mix well before transferring
28 Add cover to plate using polyester plate film
29 Vortex to mix plate
30 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
31 Transfer 96 well plate onto magnetic stand for 30 seconds and discard liquid
32 Add 200 µL of PBS Buffer to each well that contains beads
33 Vortex well and place plate on centrifuge for 2 minutes at 400RPM
34 Transfer 96 well plate onto magnetic stand for 30 seconds and discard liquid
35 Add 200µL of 10mM Ammonium Bicarbonate to each well that contains beads
36 Vortex well and place plate on centrifuge for 2 minutes at 400RPM
37 Transfer 96 well plate onto magnetic stand for 30 seconds and discard liquid
38 Add 100µL of 0.1% TFA in water to each well and vortex
39 Add pH paper to check pH value (should be lower than 3)
40 Add plate cover and shake for 10 minutes at 1200RPM using thermomixer
41 Transfer plate on centrifuge for 2 minutes at 400RPM
42 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

$853.00 (Ea)

Qty:

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Signature Peptides of Rituximab on a bioZen Peptide XB-C18 Column Using bioZen MagBeads

Order Items Used in This Application

LC Application

App ID 25617

Applications

Signature Peptides of Rituximab on a bioZen Peptide XB-C18 Column Using bioZen MagBeadsshareLinkedinprintfavorite_border

Order Items Used in This Application

LC Application

App ID 25617

Signature Peptides of Rituximab on a bioZen Peptide XB-C18 Column Using bioZen MagBeads