Anti-Dopamine抗体(ab6427)

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ab6427 is not guaranteed for use in immunohistochemistry. However, ab1001 (https://www.abcam.com/ab1001) is guaranteed to work in immunohistochemistry and gave good results when fixed with formalin/PFA in paraffin-embedded sections.

Regarding your question about whether this antibody works on rat tissue, because the immunogen of ab1001 and ab6427 is a Chemical / Small Molecule, the species in which this antibody is used is irrelevant. Therefore both these antibodies should work on rat tissue.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

We have additional information about capturing the DA as follows: After discussion with our research team, we think that the better way is to bind DA on your tissue via glutaraldehyde (0.1 to 1%). - Double bounds are reduced by adding of sodium borohydrure. - Tissue has to be wash for eliminate borohydrure excess. - You have also to make a competition scale between antibody and different concentration of the antigen DA-G-BSA. Had this medium on the tissue. - After revelation by a labelled anti-rabbit antibody, you finally have to estimate the competition rate between DA linked on tissue and soluble DA-G-BSA conjugate.

Thank you for your enquiry. For Immunohistological studies with dopamine antibodies, fixation with glutaraldehyde is required. Please find below a protocol. Perfusion protocol for adult male Sprague-Dawley (weight around 0.5 kg) 1- The animals can be deeply anaesthetized for example with uerethane (0.1-1.5g/kg, intraperitoneal). 2- Heparinized and perfused via the ascending aorta with 100 ml of cold physiologic saline (0.9% NaCl) and with the following fixative solution: a. 300 ml of cold 4% paraformaldehyde and 2% glutaraldehyde in 0.1M Phosphate buffer (PB), pH 7.2 ( in two minutes). b. 600 ml of cold 4% paraformaldehyde and 2% glutaraldehyde in 0.1M PB, pH 7.2 ( in ten minutes). c. Dissect out the brains and place in a solution of 4% paraformaldehyde in 0.1M PB, pH 7.2, at 4°C for twelve to sixteen hours. d. Before brain will be cut on a freezing microtome, we must include it in growing concentrations of sucrose (5 at 30 %). If you have any further questions then please do not hesitate to get back in touch with us.

The pump that was used is a peristaltic pump with a flow of 150-300ml/minute. GILSON supply this pump type.

This antibody is known to work with 3-5% glutaraldehyde perfusion. Protocol is as follows - Perfusion : perfuse intracardially through the aorta using a pump with the following solutions : solution A (30ml) : 200-300ml/min solution B (500ml) : 200-300ml/min Solution A : cacodylate 0.1M, sodium metabisulfite 10g/l, pH = 6.2 Solution B : cacodylate 0.1M, sodium metabisulfite 10g/l and glutaraldehyde 3-5% pH = 7.5 Post fixation : 15 to 30 min in solution B, then 4 soft washes in Tris 0.05M with sodium metabisulfite 8.5g/l, pH 7.5 (solution C). Tissue sectioning : Cryostat or vibratome sections can be used. Reduction step : Sections are reduced with the solution C containing sodium borohydride (0.1M) for 10 min. Then, the sections are washed 4 times with solution C without sodium borohydride. Application of anti-Dopamine antibodies : The final dilution is 1/2,000 to 1/5,000 in solution C containing triton X100 0.1%, plus 2% of non-specific serum. A dozen sections can be incubated with 2ml of antibody solution overnight at 4°C. Then the sections are washed 3 times (10 min) with solution C. N.B. : Antibodies may be used at a higher dilution. The customer should explore the antibody dilution to reduce the possibility of high background. Note that a substitution in the buffer system as used in our protocol may change the background and the antibody recognition. PAP procedure : Secondary antibody : Sections are incubated with 1/100 dilution of goat anti-rabbit in solution C for 3 hours at 20°C or 1 hour at 37°C. Then washed 3 times (10 min) with solution C. PAP : Sections are incubated with 1/1,000 dilution of rabbit peroxidase anti-peroxidase complex in solution C for 1 hour at 37°C. Then wash 3 times (10 min) with solution C. Detection : Antibody-antigen complexes are revealed using diaminobenzidine (25mg/100ml) (or other chromogen) dissolved in Tris 0.05M and filtrated; 0.05% of H2O2 is added. The sections are incubated for 10 min at 20°C. Reaction is stopped by transfering sections in 5ml of Tris 0.05M.