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N-terminal Edman Sequencing

N-terminal sequencing of proteins

Quick PICK ‘n POST™ N-terminal Edman sequencing of your protein – performed in 4-8 days!

Our N-terminal Edman sequencing service provides detailed information about the N-terminal amino acid sequence of your protein. It is a cyclic procedure, where amino acid residues are cleaved off one at a time. Then we identify each amino acid one-by-one by chromatography.

Service includes

  • You get the first 5, 10 or 20 N-terminal amino acids. (Scroll down this page for pricing).
  • We send the report to you within 4-8 days.

This analysis is 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.

If you have further needs

  • You want determination of up to 80 amino acids from both N- and C-terminal?
  • Looking for analysis of proteins in-solution?
  • Need separation of proteins prior to analysis?
  • You want comparison with expected amino acid sequence?

Then send us a request, so we can enter into a dialogue with you to meet your specific needs.

Services/Products

PICK ‘n POST™ N-terminal Edman determination of 5 amino acids

  • PVDF membrane bound 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
Request quote

PICK ‘n POST™ N-terminal Edman determination of 10 amino acids

  • PVDF membrane bound proteins
  • N-terminal Edman degradation
  • Determination of 10 amino acids
  • Reporting include raw data

$650.00

Fast and Easy
Will be delivered in 4-8 days
Info / Order
Request quote

PICK ‘n POST™ N-terminal Edman determination of 20 amino acids

  • PVDF membrane bound proteins
  • N-terminal Edman degradation
  • Determination of 20 amino acids
  • Reporting include raw data

$1,000.00

Fast and Easy
Will be delivered in 4-8 days
Info / Order
Request quote

Technical details

Introduction
The Edman degradation chemistry is a cyclic procedure where the PITC reagent is coupled to the free N-terminal amino group and cleaved off the protein. The PITC coupled residue is transferred to a flask, converted to a PTH-residue and the amino acid is identified by HPLC chromatography. The Edman degradation reaction cycle is repeated and a sequence of N-terminal amino acids is determined.

Since the Edman sequencing process never is 100% there will be a decrease in signal intensity as the process goes a long. Therefore the amount of material needed will increase with increased number of residues.

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

 

Sample requirements

Introduction
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 will show a mixture of amino acids in each Edman degradation cycle, and a sequence can not be determinated.

Sample Preparation
Proteins can by analyzed from PVDF membranes after electroblotting from 1D/2D gels.

PVDF membrane samples
Eletrophoresis and electroblotting
Run 1D or 2D gel
Blot onto a PVDF membrane – DO NOT use Nitrocellulose membranes!
Wear gloves and avoid dust (keratin) contamination of gel and membrane

Stain the PVDF membrane with Ponceau or Coomassie Blue
Coomassie Blue and Ponceau S work well in combination with N-terminal Edman Sequencing – DO NOT use silver staining!
Wash the PVDF membrane in water
IF glycine is used in the transfer buffer, the PVDF membrane should be washed carefully to remove the glycine
Learn here more about optimal PVDF membranes, blotting buffers and staining methods for Edman sequencing

Cut out the bands/spots of interest
Use a clean scalpel to cut out the protein bands. They should be clearly visible by Coomassie Blue/Ponceau staining to have sufficient protein amounts
Cut the protein band close to the edge to have a high protein concentration on a small membrane area
Cut and send 2-3 identical PVDF bands/spots if possible, for eventual back up analysis

Put the bands/spots into separate wells/tubes

 

Sample shipment

shipping option
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.

  • 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.
200 Page Mill Road, Suite 100
Palo Alto, CA, 94306
USA
Att: Jette Williams
Tel: (650) 543-3193
Customers outside of USA
Alphalyse A/S
Unsbjergvej 4 D
DK-5220, Odense
DENMARK
Att: Anette Rasmussen
Tel: +45 6310 6500
shipping option
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.
  • 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.

shipping option
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.
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. Fill with cooling units/cold packs.
  • 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.

shipping option
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

Read more
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