Pocono Rabbit Farm and Laboratory Inc.

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ANTIBODY PROGRAM DESIGN SERVICE- ASSISTANCE IN PEPTIDE SEQUENCE SELECTION In this FREE service,PRF&L staff uses computer algorithms to predict antigenic regions within an amino acid (or translated DNA) sequence, for those customers interested in anti-peptide antibody production. The customer will receive a minimum of 3 antigenic peptide sequences corresponding to these optimal antigenic domains, the pI, MW, and calculated peptide charge for each peptide. Also included in this service are strategies for the bioconjugation to larger carrier proteins and a suggested antibody host. Please provide your sequence to us electronically in an email. Advanced Polypeptide Characterization is Available- for specifics and pricing, please inquire.

CUSTOM PEPTIDE SYNTHESIS Please use block letters.
PEPTIDE #1:NH2- -COOH    Total amt needed
PEPTIDE #1 ID: please limit to 10 characters                 mg Purity %

PEPTIDE #2:NH2- -COOH    Total amt needed
PEPTIDE #2 ID: please limit to 10 characters                 mg Purity %

CONJUGATION OF PEPTIDES, POLYPEPTIDES, AND SMALL MOLECULES
Please indicate your conjugate choice and amount of peptide to be conjugated. We typically conjugate 1.5mg of free peptide per animal used and 1mg for ELISAs.
KLH mg | BSA mg | OVA mg | Thyroglobin mg | Albumin mg
INSTRUCTIONS:

ESTIMATING ANTIBODY TITERS by ELISA
[Enzyme Linked ImmunoSorbant Assay]
Four bleeds/samples can be titered per plate. Peptides need to be conjugated to a carrier other than was used for immunizations. Preimmune bleed is titered with the first ELISA. ELISAs are then normalized to previous bleed.

ELISA after first immune bleed.
ELISA the first and second bleed after the second immune bleed.

WESTERN BLOT ASSAY - 100ug of antigen, ~molecular weight and concentration are needed.

INSTRUCTIONS:

AFFINITY PURIFICATION OF ANTIBODIES
Antigens are covalently linked to Affinity Matrices with minimal steric hindrance, via sequence specific bio-conjugation chemistry. Where necessary, optimal binding conditions are determined to enhance antigen-antibody binding and subsequent antibody yield.
INSTRUCTIONS:

OTHER AVAILABLE SERVICES:

CUSTOM IMMUNOAFFINITY COLUMN PREPARATIONS

ENZYME AND TAG LABELING

FITC, DIG, BIOTIN, HRP, ALK PHOS, and Other Tags

RNA: TOTAL RNA ISOLATION from B – Cell Tissues for all

RNA SAMPLE ARCHIVING

CUSTOM “PHAGE DISPLAY” LIBRARY DEVELOPMENT

SDS POLYACRYLAMIDE GEL ELECTROPHORESIS - Sample run under reducing conditions or non reducing proteins are visualized by coomassie blue or silver staining methods.100ug antigen needed

ANTIGEN PURIFICATION FROM SDS POLYACRYLAMIDE GEL
Customer supplied PAGE (gel slice) embedded protein samples purified by electrophoresis.
Customer supplied mixed protein samples. Protein antigen is size fractionated on SDS PAGE gels, excised, and purified by electrophoresis.

PURIFICATION OF IgG or IgY
IgG will be purified from serum or plasma using Protein A, Protein G, or recombinant protein A/G affinity matrices.
IgY will be purified from egg yolks by the polyethylene glycol precipitation method. Protein concentration will be determined. Egg prescreening for maximum antibody yield option available. Non-PEG alternative methods and antigen specific affinity purification available.
INSTRUCTIONS:

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