N-terminal Edman sequencing

Edman degradation of your protein or antibody

We can list up to 40 amino acids of your drug substance by using N-terminal Edman sequencing (Edman degradation). Our experts perform the analysis, using the best available technology.

N-terminal Edman sequencing is a cyclic procedure. We cleave off the amino acid residues one at a time. Then we identify them by chromatography. However, the analysis will not work for samples containing multiple N-terminals and for N-terminally blocked proteins.

PROTEIN – sequencing service includes:

Optional 1D SDS PAGE or HPLC isolation of the protein.
Sequencing by Edman degradation.
Determination of up to 40 amino acids.
Comparison of data to expected drug substance sequence.

ANTIBODY – sequencing service includes:

1D SDS PAGE separation of the HC & LC.
Blotting onto PVDF membrane.
Staining and also cutting of the HC & LC bands.
N-terminal Edman sequencing – up to 40 amino acids for each chain.
Optional: De-blocking of pyroglutamic acid.

You receive the data in an easy-to-understand report.

Download application note (pdf – 1 MB). Learn more about the many applications of Edman degradtion.

Not exactly what you are looking for? Send us a request so we can adjust the analysis to meet your specific needs.

Services

N-terminal Edman determination of 5 amino acids

N-terminal Sequencing – 5 Residues

PVDF membrane bound or in solution proteins
N-terminal Edman degradation, determination of 5 amino acids
Reporting include raw data

$450.00

Fast and Easy

Will be delivered in 4-8 days

Info / Order

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N-terminal Sequencing – 10 Residues

PVDF membrane bound proteins
N-terminal Edman degradation, determination of 10 amino acids
Reporting includes raw data

$650.00

Fast and Easy

Will be delivered in 4-8 days

Info / Order

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Customized N-terminal Edman sequencing

Custom 1D PAGE, PVDF blot and band choice
Optional pGlu aminopeptidase deblocking
Edman sequencing, 10, 20 or more residues
Custom analysis for sequence variants, and semi-quantification

Customized Service

Quality analysis, presented in easy-to-understand reports

Info / Order

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Technical details

Get even more info

Sample preparation

Follow these directions

Sample shipment

Online guide

Introduction
The Edman degradtion chemistry is a cyclic procedure: It couples the PITC reagent to the free N-terminal amino group and then cleaves off the protein. Next, the PITC coupled residue is transferred to a flask and converted to a PTH-residue. Finally, the HPLC chromatography identifies the amino acid. The Edman degradtion reaction cycle is repeated until a sequence of N-terminal amino acids is determined.

Since the Edman sequencing process never is 100%, the signal intensity decreases as the process goes a long. Therefore the amount of material needed also increases with increased number of residues.

The Edman sequencing analysis will not work if the N-terminal amino group is blocked for the PITC chemistry. E.g. due to acetylation or pyroglutamic acid, which is frequent for antibodies. If the sample contains more than one protein, the chromatography then shows multiple amino acids, and a unique sequence can thus not be read.

The analyses where we find 5-10 amino acids are only performed on PVDF membrane bound proteins.
Use this guide for help on How to convert your sample to PVDF membrane bound proteins. You can also ask us to run the 1D/2D gels and transfer to PVDF membrane.

Protein sequencing analysis by N-terminal Edman degradation requires 2-10 micrograms, or 10-50 picomoles, of a pure protein. If a protein sample contains more than one protein, the analysis shows a mixture of amino acids in each Edman degradation cycle, and a sequence is therefor not determined.

Ship your protein samples by a courier company of your own choice

Place your samples into micro tubes and pack in a suitable transport box. Remember to print and include the sample submission form.

Contact a courier company of your choice (FedEx, UPS, DHL, TNT, World Courier etc.). Use the following sentence to describe the content: “Protein material – Non-toxic, non-hazardous, for research purpose only“.
Use cool packs or cooling units if your samples should be stored below 5°C/40°F
Ship the samples to the office closest to you (Canadian customers, please ship to Europe):

Customers in the USA
Alphalyse, Inc.
299 California Avenue, Ste 200
Palo Alto, CA 94306
USA
Att: Thomas Kofoed
Tel: (650) 543-3193

Customers outside of USA
Alphalyse A/S
Unsbjergvej 4 D
DK-5220 Odense SO
DENMARK
Att: Anette Rasmussen
Tel: +45 6310 6500

Order a prepaid FedEx Air Waybill < 1 lb./ 500g

A prepaid FedEx Air Waybill can be used for fast and easy shipment of samples in packages up to 1 lb./ 500g.
We will generate a prepaid Air Waybill, prefilled with the correct information, and send it to you by e-mail (PDF-format) within 24 hours.
Then follow these simple instructions:

Place your samples into micro tubes. Pack in suitable transport box. Remember to print and include the sample submission form.
Print the PDF document and stick it on the transport box or just hand it to your FedEx officer upon pickup.
Call to request a pickup from your local FedEx office.

If you plan to send your samples stored at room temperature, this option is usually the right choice. If you fill your box with cooling packs, the weight will most often exceed 1 lb./500g.

Order a prepaid FedEx Air Waybill < 10 lb./5kg (e.g. for packages with cooling packs)

This prepaid FedEx Air Waybill can be used for shipment of samples in packages up to 10 lb./ 5kg, with or without cooling packs (do not use dry ice!).
We will generate a prepaid Air Waybill, prefilled with the correct information, and send it to you by e-mail (PDF-format) within 24 hours.
Then follow these simple instructions:

Place your samples into micro tubes. Pack them in a suitable transport box. Remember to print and include the sample submission form.
Fill with cooling units/cold packs (do not use dry ice!).
Print the PDF and stick it on the transport box or just hand it to your FedEx officer upon pickup.
Call to request a pickup from your local FedEx office.

If you plan to fill your transportation box with cooling packs, select this shipping option. If you plan to use dry ice, you cannot use this Waybill!

Get a quote for shipment by World Courier

The World Courier service can be used for shipment of samples of all types. If you select this option, we will contact you to provide a quotation based on location and shipping conditions. You will receive a prepaid Air Waybill which will be invoiced together with your analysis results. This is usually a good option if FedEx does not service your location.
With the prepaid World Courier service you get:

GxP compliant processes to ensure maximal security and traceability
Door-to-door solution on a dedicated vehicle
Possibility of providing qualified packaging, temperature monitoring and dry ice
Shortest possible transit-time through “next flight out”-procedure
Customs clearance and help with paperwork