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Inquiry: We tested this antibody because it's the only one commercially avalaible which recognized Nav1.7 in human tissues and which is published. But we failed to observe any labelling in our "control" tissues where Nav1.7 is known to be expressed (fibers in dental pulp, Luo et al, 2008 fibers in skin). There is no specific immunolabelling when hNav1.7 is expressed in HEK cells. Find in the attached file more details concerning the different protocols we tested. This is very disappointing. |
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Thank you for taking time to put the information together and for contacting us. I am sorry to hear this antibody is not providing satisfactory results. |
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
ab65167 staining Nav1.7 in human sperm cells by Immunocytochemistry/ Immunofluorescence.Sperm cells were washed, re-suspended in phosphate-buffered saline and smeared onto poly-L-lysine-coated slides. Spermatozoa were then fixed by incubation in cold methanol (-20ºC) for 20 minutes. Slides were washed three times for 10 minutes with PBS and incubated with 2% BSA in PBS for 30 minutes to block non-specific sites. Test slides were incubated with ab65167 at a 1/100 dilution. Samples were washed three times in PBS, and incubated for 60 minutes with appropriate FITC-conjugated secondary antibodies. Slides were further washed in PBS, mounted using Vectashield and examined with a Olympus BX-51 fluorescence microscopy using a 100× immersion objective.
Image from Pinto FM et al, Reprod Biol Endocrinol. 2009 Jul 16;7:71, Fig 2.
ICC/IF image of ab65167 stained PC12 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab65167, 1µg/ml) overnight at +4ºC. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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