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Procedure Beckman Coulter ProteomeLabTM PA 800

Steps for sample testing using the SDS MW Method

Prepared By: Rafael Hernandez , Jan 2013

Procedure: Logging in • Turning on instrument PA800

– Refer to “Preconditioning SDS procedure”.

• Logging into windows

– Refer to “Preconditioning SDS procedure”

• Logging into the PA800 software (32 karat 8.0 software)

– Refer to “Preconditioning SDS procedure.

– Choose the SDS MW icon

Procedure: Logging into SDS MW

• On the 32 karat software window (see next slide), log in by typing the following: – Username: PA800

– Password: pa800

– Project: SDS MW (select from scroll down menu)

• If you are not in the Direct Control window, do the following: – select from the top menu, Control

– Select “Direct Control”

– Select “View” (see next slide)

Procedure: Turning on and Calibrating PDA Detector

• Turn the lamp on by clicking on the rainbow icon located on the “Direct Control” view.

• Follow the instruction on the lamp control window (see next slide) to turn it on.

• Calibrate detector by clicking on “Calibrate” on the “PDA Detector Parameters”. It will take about 2 minutes for calibration to complete. If calibration fails, make sure that the lamp remains “on” for at least 20 minutes before repeating the calibration.

Check Buffer Trays used during preconditioning and sample testing

• Go to the “Direct Control” View. – Click on “Home”. Trays-rack will move around until it position

itself in buffer vials BI:A1 and BO:A1. – Then, click on “Load” – Open the lid of the instrument, and position the buffer vials as

indicated in the next two slides (layout of buffer trays and vials). • The first slide lay out is for 6 injections or cycles. If planning to do

more than 6 cycles, an additional row of buffer and gel vials is needed for every 6 cycles.

• The second slide lay out is for 24 injections or cycles.

– Close the lid. Ensure that is completely close otherwise the trays will not move to the “home” position (TI:A1 and TO:A1). • It may be necessary to push down the lid and the front cover where

the capillary cartridge is located.

vial H2O

vial Empty

vial Gel

vial NaOH

vial HCl

vial H2O

vial H2O

vial Waste

vial Gel

vial Waste

vial Waste

vial Waste

A B C D E F

Inlet Buffer (BI) Tray on the left Outlet Buffer (BO) Tray on the right

A B C D E F

Layout of the Buffer Trays/Vials during preconditioning and for a CE run of 6 injections (cycles) max

vial H2O

vial H2O

vial Empty

vial Gel

vial NaOH

vial HCl

vial H2O

vial H2O

vial Empty

vial Gel

vial NaOH

vial HCl

vial H2O

vial H2O

vial Empty

vial Gel

vial NaOH

vial HCl

vial H2O

vial H2O

vial Empty

vial Gel

vial NaOH

vial HCl

vial H2O

vial H2O

vial H2O

vial Waste

vial Gel

vial Waste

vial Waste

vial Waste

vial H2O

vial Waste

vial Gel

vial Waste

vial Waste

vial Waste

vial H2O

vial Waste

vial Gel

vial Waste

vial Waste

vial Waste

vial H2O

vial Waste

vial Gel

vial Waste

vial Waste

vial Waste

A B C D E F

Inlet Buffer (BI) Tray on the left Outlet Buffer (BO) Tray on the right

A B C D E F

Layout of the Buffer Trays/Vials during preconditioning and for a CE run of 24 injections (cycles) max

Procedure: Performing Pre-conditioning

• Perform preconditioning before CE sample testing.

– For preconditioning refer to “Preconditioning SDS_procedure”.

Procedure: Performing CE sample testing using the SDS MW method

• Prepare sample(s) as indicated in “SDS-MW Sample Prep Procedure”.

• For a 24 sample injection or cycle, set the sample vials into the “Inlet Sample Tray” as indicated in the next slide from position A1 (sample 1) to C8 (sample24). The Inlet Sample Tray is located in the back of the instrument sample tray. – For loading sample vials into the instrument follow

“Preconditioning SDS procedure” section called “Check Buffer Trays used during preconditioning”.

Sample 8

Sample 16

Sample 24

Sample 7

Sample 15

Sample 23

Sample 6

Sample 14

Sample 22

Sample 5

Sample 13

Sample 21

Sample 4

Sample 12

Sample 20

Sample 3

Sample 11

Sample 19

Sample 2

Sample 10

Sample 18

Sample 1

Sample 9

Sample 17

A B C D E F

Inlet Sample (IS) Tray located on the left buffer/sample compartment

Layout of sample vials for CE sample run

8 7 6 5 4 3 2 1


• Write a sequence.

– In the direct control view, go to “file”, then “sequence”, then “open”, then “SDS20.seq”, and click on “Open” (see next three slides).

• The sequence will open, and must be saved as a different file (ex. SDSMW_date_operator initials)


• Write a Sequence (cont’d). On the “Sequence” page, the headings of the chart are the following: – “Run #” indicates the sample injection or cycle for

preconditioning.

– In a 24-sample or cycle format, • Run # 1, 8, 15 and 22 are set up for preconditioning the

capillary every six sample injections. Ensure that the “Method” associated with preconditioning is “20cmPreConditioning.met”.

• Run # 2-7, 9-14, 16-21, and 23-28 are set up for samples and SDS-MW analysis. Ensure that the “Method” associated with these injections is “SDS20Separation.met”.


• Write a Sequence (cont’d). – The “Sample Inject Inlet” heading indicates the

position of the sample vial for that run. • For example, for “Run # 2”, the nomenclature “SI:A1” means

sample inlet tray, vial position A1. • Make the necessary changes on the chart that will match the

position of each of your sample vials.

– The “Sample Inject Outlet” heading indicates the outlet of the capillary during that cycle. • For example, for “Run # 2”, the nomenclature “BO:C1”

means buffer outlet tray, vial position C1. • Make the necessary changes on the chart that will match the

position of each of your vials.


• The next eight slides contain the program graphical screen for each step during the set up of a sequence.

Sequence Page


• Once the “Sequence” is set up, instrument is ready for the CE run.

– Go to the sequence page, top menu and select “Control”.

– Under “Control” select “Sequence run”.

– A “Run Sequence” window will appear, then click on “Start”.

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