Opsin 1 (Medium Wave) Antibody - BSA Free

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• Opsin 1 (Medium Wave) Antibody

  Name: Anonymous

  Application: Immunohistochemistry-Frozen

  Sample Tested: mouse retinal tissues

  Species: Mouse

  Verified Customer | Posted 12/27/2016

  An image of retinal cryosection that I tried to label with the same antibody by Novus, but I did not get any signal from photoreceptor outer segments where the positive staining should be localized.

  Cryosection Primary: Chicken anti M-opsin 1:50 Secondary: Goat anti-chicken CY3 secondary 1:100 Procedure 1. Take the sections out from freezer and thaw the slides at room temperature for 10 minutes 2. Surround the tissues with a hydrophobic barrier (proper big lines!) using a barrier marker pen a. Let to dry properly 3. Rinse the sections in 2X in PBS (5 min each) 4. Drain the excess wash buffer 5. Incubate the sections in blocking buffer (10 % NGS) for 1 h at RT a. 50 µl of solution 6. Drain the excess blocking buffer 7. Incubate in primary antibody overnight at 4º C in a wet chamber. a. 0.01 M PBS + 1 % NGS as buffer b. 50 µl of solution 8. Next day rinse the sections in PBS+triton (0.25 %) for three times (5 min each) 9. Incubate with Secondary antibody prepared in PBS for 2 hrs. a. 50 µl of solution 10. Rinse the sections in PBST twice (5 min each) 11. Rinse the sections in PBS twice (5 min each) 12. Drain the excess wash buffer a. Air-dry for few mins 13. Mount with an anti-fade mounting media 14. Visualize using a fluorescence microscope

  

• Opsin 1 (Medium Wave) Antibody

  Name: Anonymous

  Application: Immunohistochemistry-Whole Mount

  Sample Tested: mouse retinal tissues

  Species: Mouse

  Verified Customer | Posted 12/27/2016

  Below is a figure of retinal whole mount stained by anti S-cone antibody from Merck Millipore and anti M-cone antibody by Novus. Millipore’s antibody stains cone photoreceptors distinguishly, but Novus’ does not.

  Tested in an untested application (IHC-whole mount) and earned the Innovator's Reward for this product. Immunostainning of the retinal flat mount Prim. antibodies: Anti M/L-cone opsin, chicken polyclonal, 1/50 dil., 1/100 dil. & 1/200 dil. Sec. antibodies: Goat anti-chicken CY3, 1/100 dil. 1. Fix the eye for 3 hours 2. Dissect retina from the eye 3. Wash overnight in PBS 4. Wash the tissue in PBST 2 times; 10 min each a. Triton-X 0.5 % 5. Block the whole mount in 10% normal goat serum (NGS) solution in PBST (1 hr) 6. Incubate o/n in primary antibody solution at 4 º C, primary ABs made in 1X PBST (0.05M) + 1% NGS 7. Next day wash in PBST + 1 % NGS; 3 X 10 min each 8. Incubate in secondary antibodies for 2-3 h a. Protect from light with tin 9. Wash in PBST + 1 % NGS; 2 X 10 min each 10. Wash once in PBS for at least 5 min 11. Transfer the whole mount on the microscope slide onto a slide retina side up. 12. Remove PBS, add mounting medium. 13. Place coverslip. Observe. If correct… 14. Secure with nail polish Samples are C57BL retinas

  

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