Cancer Reagent Target Request Form

A field with an asterisk (*) before it is a required field.

* Investigators submitting must demonstrate that the target request is consistent with the mission of the NCI and that the target reagent will be further evaluated once created (i.e. NIH funded research or equivalent):

For each question, please answer in 500 words or less:

Antibody

1. * Describe why the antibody is required, the importance of the target and the research area to which it would benefit if the material was produced (do not include confidential/proprietary information).
2. * Describe commercial antibody availability or lack thereof and any experience you have had with these antibodies.
3. * Describe the intended application(s) for the antibody (i.e. Western Blot, ELISA, IHC):
4. * Describe any recommendations for positive/negative controls (i.e. Cell lines to be used) that would be useful in screening the antibody supernatants to aid in the selection of the final antibody(ies) for the project proposed. Does your lab possess/have access to the controls you recommend?

Antigen

1. * Describe the molecular weight of the antigen. A full-length protein is desired, when possible, in order to create reagents which will bind native conformations. However, functional domain/protein fragments or peptides of interest will be considered.
2. * Where was the antigen produced? What was the expression system (i.e. E.coli, mammalian, baculovirus, other)?
3. * Provide the antigen amino acid sequence (do not provide confidential/proprietary information)?
4. * How much protein/peptide will you provide for this project? Please note that a minimum of 3- 5 mg of protein/peptide is required for monoclonal antibody production, screening, counter screening (where applicable) and characterization (preferably endotoxin free) is required. If a different protein/peptide is needed for counter screening and/or characterization you will need to provide that as well (1 – 3 mg).
5. * For monoclonal antibody generation, is the protein/peptide endotoxin free? How was this determined? If needed, NCI's Antibody Characterization Laboratory will provide endotoxin removal to accepted targets, but more than 5 mg of protein may be needed since the removal process often leads to a loss of 10–50% of protein.
6. * If you are using a peptide for the immunogen are you targeting any post-translational modifications such as phosphorylation and if so, please indicate on the peptide sequence which specific site(s).
7. * Is the protein/peptide soluble in a non-denaturing buffer (i.e. PBS)? Soluble proteins are desired, but insoluble proteins will be accepted.
8. * Provide copies of gels or other characterization(s) that have been performed on the antigen to demonstrate purity, molecular weight, etc. Additionally, proteins received will be subjected to QC evaluation to access purity, concentration and identification (SDS-PAGE, MALDI-TOF) and those which do not meet specifications will be rejected.
   Send attachments to: hiltket@mail.nih.gov
   
   
9. * Provide any relevant publications regarding the protein/peptide.

*Please read and indicate if you agree with the following statements:

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