ANTI-COLLAGEN I ALPHA I CHAIN PROPEPTIDE

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 PR845, anti-collagen I alpha I chain propeptide antibody recognizes the C-terminal propeptide of collagen I alpha I.  Because this sequence is lost during the maturation of fibrillar collagen, this antibody binds only to collagen molecules that are still associated with the cell in which they were synthesized.  Therefore, this antibody can be used to identify cells in which collagen I alpha I is being expressed.  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 human and mouse samples.  Based on the similarity of the immunogenic human peptide with homologous sequences in the collagen I alpha I chains of several species, this antibody is expected to recognize the collagen I alpha I polypeptide in most, if not all, species of mammals, 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:

0.1  to 0.2 micrograms per ml for Western blots.

2 to 5 micrograms per ml for immunohistochemistry