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Analysis of N-terminal Amino Acid Sequence

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Product Description

Analysis of N-terminal amino acid sequence (protein sequencing) is a method to excise amino acid one by one from N-terminal of protein or peptide (Edman degradation method) to be segregated by HPLC, then to detect by UV to determine amino acid sequence by the time the peak is gained (Chromatogram).

Analysis

  • 1 Amino acid sequence analyzer is PPSQ-33A.
  • 1 Residues possible to be analyzed is 5 to 20 residues (depending on the samples).
  • 1 Analysis will be discontinued if it was not possible due to N-terminal block or lack of sample molar mass etc. Even in that case, data up to 5th residue will be taken and the analysis basic fee will be charged.
  • 1 Please note that when data was not processed because of the N-terminal block etc., there will be no analysis retry. Request for the analysis of the same sample will be accepted as a new order.

Deliverables

Amino acid sequence identified by chromatogram, or comment in case analysis was not possible, together with a raw chromatogram data.

Price and Domestic Lead-Time

<Price>

Normal Analysis Rapid Analysis
Basic Fee 50,000 JPY / sample
Including analysis fee for up to 5 residues		 50,000 JPY / sample + 15,000 JPY / sample (rapid fee)
Including analysis fee for up to 5 residues* Maximum sample numbers we can accept for rapid analysis per one order are 3 samples. Please refer to the details below.Analysis residues      Sample numbers

10 residues or less 3
between 11 and 20 residues 2
Analysis Fee 6,000JPY per residue
Charged for analysis of more than 6 residues.
When analysis was discontinued for 3 or less residues by the judgement that it was not possible, analysis fee will not be charged.
Sample Preparation Fee For liquid or dried samples:
morecular weight of 10,000 or more: 20,000 JPY / sample
morecular weight less than 10,000: 30,000 JPY / sample
* Please send information which shows sample purity.e.g. SDS-PAGE, ChromatogramFixing to PVDF membrane:

Basic charge: 50,000 JPY
Preparation: 7,000 JPY / sample
Protein having a molecular weight of 10,000 or more is acceptable.
For protein having a molecular weight less than 10,000, please inquire.
* Please attach a SDS-PAGE image.
Other Service Charges Please inquire
Domestic Lead-Time Within 10 business days from sample acceptance.

* Actual schedule will be informed.

* It may take more depend on sample numbers and analysis conditions.

* Orders from sample preparation would require extra business days (from one to two days depend on sample numbers).

 Within 5 buiness days from sample acceptance.

* Orders from sample preparation would require extra one business day.

 

Regarding Sample

Sample preparation

Required sample:
PVDF membrane
* In case of liquid or dried samples, an additional fee for sample preparation will be required.
For details, please inquire.

Sample amount
For the amount of sample, i.e. the number of lanes, required for analysis, please refer to the
below images for your information.
* Transcribed PVDF membrane was stained with CBB after SDS-PAGE using 10 wells mini gel.

We will discuss with customer based on the PVDF membrane or the SDS-PAGE image submitted with an order sheet.
* Please be noted that orders may not be acceptable depend on the amount and purity
of protein.

 

Sample status

  • 1 If PVDF membrane : 3 to 4 membranes of 1 mm x 8 mm
    Recommended stain method : CBB-R250 staining, Ponceau S staining, Amido Black staining
    * Large amount of CBB may cause some peaks around PTH-Met, Val or Pro on chromatogram and disturb PTH amino acid detection.
    * Please do not use Hybond-P or nitrocellulose membrane.
  • 1 If liquid: Please prepare for 100 ul or more sample at a concentration of 5 pmol/ul or more.
    Please refer to Allowable Value of Harmful Substance In Amino Acid Sequence Analysis
  • 1 If in a dry form (powder)
    * In case sample was liquid or dried form, additional sample preparation fee will be charged.
    * Please attach an Image of the sample after blotting.

Handling of Sample

Shipment condition
  • 1 If PVDF membrane or dry form(powder) : Please use refrigerated courier of 4°C
  • 1 If liquid : Please use refrigerated courier of – 20°C(packed with enough dry ice).
Acceptance QC of samples

We will discuss with customerabout the PVDF membrane or SDS-PAGE image submitted
by e-mail.
Any sample that was determined not to be possible to analyze is to be returned to customer at customer expense or discarded.

Left over samples

After analysis, left-over sample will be discarded.

Please refer to Allowable Value of Harmful Substance In Amino Acid Sequence Analysis

 

Ordering

Please inquire for details.

Solvent for the N-terminal amino acid analysis orders in liquid form

If liquid: Please make sure to use adequate solvents as below. If in a dry form, please also make sure that the solvents other than volatiles are adequate.

adequate solvents

* distilled water
* TFA solution
* solution of acetate
* acetonitrile, methanol (15% or less)
* trimethylamine solution

 

inadequate solvents

* Ammonium salts
* Tris and the like
* Solvent containing primary and secondary amine
* Solvent containing large amount of surfactant such as salt and SDS
* Solvent containing highly polar substances such as Glycerol, reductant and urea
* Solvent containing involatiles

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