Download The First Class Program

>
First-Class Program Evaluation Form
The First Class Program
Your purchase order reference:
EGT Quotation Number:
Antibodies that work in your application, guaranteed
> Custom peptide antibodies adapted to
VAT Nr. :
If VAT exempt please include a copy of certificate
DELIVERY ADDRESS
INVOICE ADDRESS
(Please fill in delivery information if different from invoicing information)
Institute/Company:
Institute/Company:
Department/Lab.:
Department/Lab.:
Contact Person:
Contact Person:
Address:
Address:
your application
> B-Epitopes are selected based on the
antigen’s predicted 3D structural
Postal Code:
features in your application conditions
Country:
Country:
Phone:
Phone:
Fax:
Fax:
e-mail:
e-mail:
> The right antibody for your immunoassay
or your money back
We have improved antigenic sequence prediction
algorithms
The antigenic surface of a protein is a 3-D patchwork of
small linear antigenic sequences. Individually, each
sequence is antigenic but, depending on the number of
accessible residues in the interaction, the affinity will be
either acceptable or too low. Currently, antibody affinities
of 10-8 to 10-9 M can be expected for linear epitope, by
calculating and analyzing the structure of each potential
epitope. We will propose the most appropriate antigenic
fragments that should exhibit such affinities.
Ref: PRO-FIRSTCLASS-0207-V2
An epitope for each application
As the structure of your protein may vary with each assay,
an assay–antigen specific analysis must be made to
determine structural and interaction properties. Major
factors leading to changes in the epitope structure are (1)
denaturation by detergent and (2) chemical modification
of the amino acids of the epitope, for example with fixatives
or other chemicals. For your immuno-assay technique
the most exposed epitope may not be your best choice.
Depending on whether the antibody will be used on a
native, a detergent-denatured or a fixative-modified
structure of your protein, we shall select either a flexible
or a more rigid region of the protein such as a turn or a
helix. Increased flexibility is reported to lead to improved
affinity, however, rigid structures can provide good
epitopes more resistant to detergent denaturation.
Additionally, sequences containing aromatic residues
rather than highly polar amino acids may be selected if
application conditions will involve the use of chemicals
that react with polar moieties. Indeed, as long as they do
not impair the fragment accessibility, hydrophobic
residues could improve the affinity. The prediction of the
best epitope is closely linked to the application for which
the antibody will be used.
The “First Class” program – what do we do?
Using a consensus of epitope prediction procedures we
characterize all potential linear epitopes. Then, in a
second step, we calculate their structures and keep those
whose characteristics are adequate to raise high affinity
antibodies. In the third step, we select the fragment(s) that
are the most likely to be useful in your immuno-assay.
What do we need ?
The accession number of the protein of interest and a
complete description of the application in which you will
use the antibody. Please complete the questionnaire on
the reverse side.
We include
•
•
•
•
•
•
•
•
•
•
Application specific peptide design
5 pre-immune rabbit sera for animal selection
Synthesis of 2 peptides, 20-25 mg each
Coupling of each peptide to a carrier protein
Double X protocol in a 4-month program
ELISA testing to monitor titer evolution
Affinity purification of 5 ml of serum against each peptide
Delivery of 15 - 20 mg of free peptide
Remaining antisera
All shipping costs
City:
Postal Code:
Protein accession #:
1. For which experimental method do you want a guarantee?
(only one technique please)
[email protected] • [email protected] • www.eurogentec.com
5. Is your protein a membrane protein
Yes
ELISA in the absence of detergent or fixative
Western Blot
Immunoprecipitation with no detergent
Immunoprecipitation in the presence of detergent
Specify the concentration and the name of the detergent
………………………………………………………………...........……………………
Staining of chemically fixed tissues
Specify the fixative ……….....................................……………...……
Staining of frozen-fixed tissues
Other technique, please specify
……………………………………………………………......……………………………
……………………………………………………………......……………………………
2. Are you working with:
Isolated cells
Other ………….............................
Subcellular fractions
3. Is your protein post-translationally modified?
Yes
No
Not known
If yes, please indicate the amino acid number and the corresponding
type of modification:
……………..…………………………………………………......……………………………
……………..…………………………………………………......……………………………
……………..…………………………………………………......……………………………
……………..…………………………………………………......……………………………
4. Are you working under denaturing conditions?
LIEGE Science Park • 4102 Seraing • BELGIUM
RPM Liège • VAT (BE) - 0427.348.346
Tel.: +32 4 372 76 65 • Fax: +32 4 264 07 88
City:
Yes
No
Not known
6. Have you already tried an immunization experiment for this
protein?
Yes
No
If yes, which peptides/protein sequences were used ?
……………..…………………………………………………......……………………………
……………..…………………………………………………......……………………………
……………..…………………………………………………......……………………………
……………..…………………………………………………......……………………………
In which type of experiments was the antibody used?
……………..…………………………………………………......……………………………
……………..…………………………………………………......……………………………
……………..…………………………………………………......……………………………
……………..…………………………………………………......……………………………
What were the results ?
……………..…………………………………………………......……………………………
……………..…………………………………………………......……………………………
……………..…………………………………………………......……………………………
……………..…………………………………………………......……………………………
7. Remarks:
……………..…………………………………………………......……………………………
……………..…………………………………………………......……………………………
……………..…………………………………………………......……………………………
……………..…………………………………………………......……………………………
……………..…………………………………………………......……………………………
No
Fax the completed order form to + 32 4 264 07 88
LIEGE Science Park • 4102 Seraing • BELGIUM • Tel.: + 32 4 372 76 65 • Fax: + 32 4 264 07 88
RPM Liège • VAT (BE) - 0427.348.346 • [email protected] • [email protected] • www.eurogentec.com
>
First-Class Program Evaluation Form
The First Class Program
Your purchase order reference:
EGT Quotation Number:
Antibodies that work in your application, guaranteed
> Custom peptide antibodies adapted to
VAT Nr. :
If VAT exempt please include a copy of certificate
DELIVERY ADDRESS
INVOICE ADDRESS
(Please fill in delivery information if different from invoicing information)
Institute/Company:
Institute/Company:
Department/Lab.:
Department/Lab.:
Contact Person:
Contact Person:
Address:
Address:
your application
> B-Epitopes are selected based on the
antigen’s predicted 3D structural
Postal Code:
features in your application conditions
Country:
Country:
Phone:
Phone:
Fax:
Fax:
e-mail:
e-mail:
> The right antibody for your immunoassay
or your money back
We have improved antigenic sequence prediction
algorithms
The antigenic surface of a protein is a 3-D patchwork of
small linear antigenic sequences. Individually, each
sequence is antigenic but, depending on the number of
accessible residues in the interaction, the affinity will be
either acceptable or too low. Currently, antibody affinities
of 10-8 to 10-9 M can be expected for linear epitope, by
calculating and analyzing the structure of each potential
epitope. We will propose the most appropriate antigenic
fragments that should exhibit such affinities.
Ref: PRO-FIRSTCLASS-0207-V2
An epitope for each application
As the structure of your protein may vary with each assay,
an assay–antigen specific analysis must be made to
determine structural and interaction properties. Major
factors leading to changes in the epitope structure are (1)
denaturation by detergent and (2) chemical modification
of the amino acids of the epitope, for example with fixatives
or other chemicals. For your immuno-assay technique
the most exposed epitope may not be your best choice.
Depending on whether the antibody will be used on a
native, a detergent-denatured or a fixative-modified
structure of your protein, we shall select either a flexible
or a more rigid region of the protein such as a turn or a
helix. Increased flexibility is reported to lead to improved
affinity, however, rigid structures can provide good
epitopes more resistant to detergent denaturation.
Additionally, sequences containing aromatic residues
rather than highly polar amino acids may be selected if
application conditions will involve the use of chemicals
that react with polar moieties. Indeed, as long as they do
not impair the fragment accessibility, hydrophobic
residues could improve the affinity. The prediction of the
best epitope is closely linked to the application for which
the antibody will be used.
The “First Class” program – what do we do?
Using a consensus of epitope prediction procedures we
characterize all potential linear epitopes. Then, in a
second step, we calculate their structures and keep those
whose characteristics are adequate to raise high affinity
antibodies. In the third step, we select the fragment(s) that
are the most likely to be useful in your immuno-assay.
What do we need ?
The accession number of the protein of interest and a
complete description of the application in which you will
use the antibody. Please complete the questionnaire on
the reverse side.
We include
•
•
•
•
•
•
•
•
•
•
Application specific peptide design
5 pre-immune rabbit sera for animal selection
Synthesis of 2 peptides, 20-25 mg each
Coupling of each peptide to a carrier protein
Double X protocol in a 4-month program
ELISA testing to monitor titer evolution
Affinity purification of 5 ml of serum against each peptide
Delivery of 15 - 20 mg of free peptide
Remaining antisera
All shipping costs
City:
Postal Code:
Protein accession #:
1. For which experimental method do you want a guarantee?
(only one technique please)
[email protected] • [email protected] • www.eurogentec.com
5. Is your protein a membrane protein
Yes
ELISA in the absence of detergent or fixative
Western Blot
Immunoprecipitation with no detergent
Immunoprecipitation in the presence of detergent
Specify the concentration and the name of the detergent
………………………………………………………………...........……………………
Staining of chemically fixed tissues
Specify the fixative ……….....................................……………...……
Staining of frozen-fixed tissues
Other technique, please specify
……………………………………………………………......……………………………
……………………………………………………………......……………………………
2. Are you working with:
Isolated cells
Other ………….............................
Subcellular fractions
3. Is your protein post-translationally modified?
Yes
No
Not known
If yes, please indicate the amino acid number and the corresponding
type of modification:
……………..…………………………………………………......……………………………
……………..…………………………………………………......……………………………
……………..…………………………………………………......……………………………
……………..…………………………………………………......……………………………
4. Are you working under denaturing conditions?
LIEGE Science Park • 4102 Seraing • BELGIUM
RPM Liège • VAT (BE) - 0427.348.346
Tel.: +32 4 372 76 65 • Fax: +32 4 264 07 88
City:
Yes
No
Not known
6. Have you already tried an immunization experiment for this
protein?
Yes
No
If yes, which peptides/protein sequences were used ?
……………..…………………………………………………......……………………………
……………..…………………………………………………......……………………………
……………..…………………………………………………......……………………………
……………..…………………………………………………......……………………………
In which type of experiments was the antibody used?
……………..…………………………………………………......……………………………
……………..…………………………………………………......……………………………
……………..…………………………………………………......……………………………
……………..…………………………………………………......……………………………
What were the results ?
……………..…………………………………………………......……………………………
……………..…………………………………………………......……………………………
……………..…………………………………………………......……………………………
……………..…………………………………………………......……………………………
7. Remarks:
……………..…………………………………………………......……………………………
……………..…………………………………………………......……………………………
……………..…………………………………………………......……………………………
……………..…………………………………………………......……………………………
……………..…………………………………………………......……………………………
No
Fax the completed order form to + 32 4 264 07 88
LIEGE Science Park • 4102 Seraing • BELGIUM • Tel.: + 32 4 372 76 65 • Fax: + 32 4 264 07 88
RPM Liège • VAT (BE) - 0427.348.346 • [email protected] • [email protected] • www.eurogentec.com