PERIOSTIN C-TERMINAL ANTIBODY

FORMAT.  Affinity purified rabbit IgGs

FORM, REHYDRATION, and STORAGE.

Lyophilized IgG in PBS (from 50 microliters of a 1 mg per ml solution). Stable for a minimum of 1 year. No preservatives added. Non-sterile. Store at 4ºC until rehydrated.  To rehydrate, add 50 microliters of water or 50 % glycerol.  Glycerol is preferred because the sample can then be stored at -20º C without freezing, thus eliminating the loss of activity that can occur during repeated freeze-thaw cycles.  At working dilutions, the level of glycerol is too small to affect most assays.  If the user prefers to store the sample at 4º C, protection against microbial growth can be provided by 50 % glycerol, 0.01% thimerosal, or sodium azide.  (If sodium azide is used, it is important that blots or slides be washed extensively after incubation with the antibody because residual sodium azide can inhibit the peroxidase activity of peroxidase-conjugated antibodies.)

SPECIFICITY.

The PR849 periostin C-terminal antibody was made against a bacterial fusion protein equivalent to an approximately 200-amino acid polypeptide from the C-terminal region of mouse periostin which is comprised of six small alternatively-spliced exons.  Consistent with the fact that this antibody is made against an alternatively spliced region of periostin, this antibody recognizes a subset of the forms of periostin recognized on Western blots by the PR847 pan periostin antibody and also shows a distinctive staining pattern by immunohistochemistry.  The antibody works on Western blots, even of crude tissue homogenates.  The antibody also works on immunohistochemistry of paraformaldehyde-fixed sections with a simple antigen-retrieval protocol (incubate slides for 20 minutes at 90º C in 10 mM sodium citrate (pH 6.0)/ 0.1 % Tween-20).  This antibody has already been shown to work on mouse and human samples and is expected to work on all mammalian species.  It remains to be determined whether this antibody works on samples from birds, amphibians, and fishes.

SUGGESTED WORKING DILUTIONS.

Different assay conditions require that serial dilution of all reagents be performed to determine optimal working concentrations. Prepare working dilution immediately before use. Storage of working dilution may result in performance loss. Suggesting starting dilutions are as follows:

to 0.2 micrograms per ml for Western blots.

2 to 5 micrograms per ml for immunohistochemistry.