Two rabbits (A and B) were immunized with a mix of two synthetic peptides of murine PTF1a sequence (one N-term peptide, KSFDNIENEPPFEFVS (aa 309-324) and one C-term peptide, EDFFTDQSSRDPLED (aa 24-38)) conjugated to keyhole limpet haemocyanin. C-term and N-term peptides have 100 % identity with rat sequence and 93% with human sequence.
We call our antibodies 7A (from rabbit A) and 7B (from rabbit B).
We designed Biacore experiments with immobilized peptides to test their specificity against immunogenic peptides (see Figure 1). 7A recognizes C-term sequence better than N-term sequence. 7B recognizes C-term sequence but not N-term sequence (see Figure 1).
Antibody 7B was purified against the « wrong peptide, N-term » but the flow through fraction has been kept. It has been tested and shows preserved immunoreactivity against C-terminal sequence (see Figure 1) and detects PTF1a in Western-Blot (see Figure 3).
We successfully used antibodies 7A and 7B in immunohistochemistry on human and mouse pancreas (see publication 1 and 3 and also see immunofluorescence in Figure 2), in immunocytochemistry of rat pancreatic acinar cells (see publication 1), Western-blot analysis of nuclear and cytoplasmic extracts of rat pancreatic acinar cells (see publication 1), of murine pancreatic acinar protein extracts (see publication 2), of transfected HEK cells (see publication 2), and in immunoprecipitation experiments (see publication 2).
Antibodies 7A and 7B are specific of pancreatic acinar cells and do not recognize any other proteins in Western-blot analysis of pancreatic ductal cells or COS cells (see Figure 3). Figure 3 also shows the absence of non specific bands.
We can provide around 30 ml of 7A antibody, 2 ml of unpurified 7B and 6 ml of 7B flow-through.
We also have lesser quantities of intermediate bleeds. Their specificity towards peptide antigens was also tested by Biacore, we can provide these results on demand in case of interest. We did not tested them in WB and IF/IHC.