Human biopsy specimens and mouse tissues were homogenized in ice-cold hypotonic lysis buffer to obtain cytosolic extracts according to a method previously published by our group.63 (link) Extracts underwent electrophoresis through a polyacrylamide minigel. Proteins were transferred onto a nitrocellulose membrane that was saturated with nonfat dry milk and then incubated with either rabbit anti-TRPV1 (Santa Cruz Biotechnology, Santa Cruz, CA), rabbit anti-TRPV4 (Novus Biological, Ltd, Cambridge, UK), rabbit anti-PPARα (Abcam, Cambridge, UK), or mouse anti–β-actin (Santa Cruz Biotechnology). Membranes then were incubated with the specific secondary antibodies conjugated to horseradish peroxidase (Dako, Milan, Italy). Immune complexes were shown by enhanced chemiluminescence detection reagents (Amersham Biosciences, Milan, Italy). Blots were analyzed by scanning densitometry (GS-700 imaging densitometer; Bio-Rad, Milan, Italy). Results were expressed as optical density (arbitrary units; mm2) and normalized on the expression of the housekeeping protein β-actin.
Rabbit anti trpv1
Manufactured by Santa Cruz Biotechnology
Sourced in United Kingdom
Rabbit anti-TRPV1 is a primary antibody that specifically recognizes the Transient Receptor Potential Cation Channel Subfamily V Member 1 (TRPV1) protein. TRPV1 is a cation channel that is sensitive to heat, protons, and certain vanilloid compounds. This antibody can be used for the detection and analysis of TRPV1 expression in various cell and tissue samples using techniques such as Western blotting, immunohistochemistry, and immunocytochemistry.
Automatically generated - may contain errors
Lab products found in correlation
Mouse anti neun, by Merck Group (2 mentions)
Donkey anti mouse cy2, by Jackson ImmunoResearch (2 mentions)
Donkey anti rabbit cy3, by Jackson ImmunoResearch (2 mentions)
Paraformaldehyde (pfa), by Merck Group (2 mentions)
Dapi dilactate, by Thermo Fisher Scientific (2 mentions)
Gs 700 imaging densitometer, by Bio-Rad (1 mentions)
Enhanced chemiluminescence detection reagent, by Cytiva (1 mentions)
Specific secondary antibodies conjugated to horseradish peroxidase, by Agilent Technologies (1 mentions)
Mouse anti β actin, by Santa Cruz Biotechnology (1 mentions)
Rabbit anti trpv4, by Novus Biologicals (1 mentions)
Rabbit anti pparα, by Abcam (1 mentions)
Cy2 donkey anti rabbit, by Jackson ImmunoResearch (1 mentions)
Donkey anti rat cy3, by Jackson ImmunoResearch (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Triton, by Merck Group (1 mentions)
Mouse anti iqgap1, by Santa Cruz Biotechnology (1 mentions)
4 protocols using rabbit anti trpv1
1
Western Blot Analysis of TRPV, PPARα Proteins
2
Immunostaining of DRG Neuron Development
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DRG neurons of different time of growth (from 3 to 12 DIV) were fixed for 20 min with 4% paraformaldehyde (Sigma) rinsed in PBS (0.01 M, pH 7.4) at room temperature, followed by washes with PBS, blocking in 5% BSA (Sigma) with 0.5% of triton (Sigma), and incubation with primary antibodies overnight at 4℃ in 1% BSA with 0.05% of triton. Following day, the primary antibodies were washed out by 1 × PBS and followed by incubation with Cy2, Cy3-conjungated secondary antibodies (Jackson ImmunoResearch) for 2 h at Troom in dark.
Primary antibodies: goat anti-Nav1.8 (1:50, Santa Cruz),22 (link) rabbit anti-TRPV1 (1:50, Santa Cruz),23 (link)–25 (link, link) rabbit anti-TRPM8 (1:50, Santa Cruz), rat anti-CD77 (1:10, Abcam),26 (link) mouse anti-NeuN (1:2000, Millipore),27 (link) and rabbit anti-Tuj1 (β3 Tubulin, 1:50, Santa Cruz).28 (link)Secondary antibodies: Cy2-donkey anti-rabbit, Cy2-donkey anti-mouse, Cy3-donkey anti-rat, and Cy3-donkey anti-rabbit (1:200, all from Jackson ImmunoResearch). DAPI (dilactate; 300 nM, Invitrogen) was used for counterstaining.
Primary antibodies: goat anti-Nav1.8 (1:50, Santa Cruz),22 (link) rabbit anti-TRPV1 (1:50, Santa Cruz),23 (link)–
3
Immunohistochemistry of Murine DRG Tissues
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Mice were transcardially perfused with PBS and 4% paraformaldehyde (PFA). Lumbar DRG (L4-L5), sciatic nerves and skin were then isolated and post-fixed in 4% PFA followed by cryopreservation in 30% sucrose. Tissues were embedded in OCT medium (Tissue-Tek) and sectioned in a cryostat at 12 μm thickness. DRG sections were placed onto poly-lysine-coated slides for immunohistochemistry. DRG tissue sections were first blocked in 5% Donkey serum plus 0.03% Triton X-100 at room temperature for 30 min. They were then incubated with primary antibodies [mouse anti-IQGAP1 (Santa Cruz), rabbit anti-TRPA1 (Alomone), rabbit anti-CGRP (Merck) and rabbit anti-TRPV1 (Santa Cruz)] at 4°C overnight. After washing in PBS, DRG sections were next incubated with fluorescence-conjugated secondary antibodies [Alexa Fluor 594 conjugated donkey anti-mouse, Alexa Fluor 488 conjugated chicken anti-rabbit and FITC-conjugated Isolectin B4 (Merck)] at room temperature for 2 h. After a thorough wash, DRG slides were sealed by coverslips with Mowoil solution.
4
Immunohistochemical Analysis of Dorsal Root Ganglia
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Samples of whole DRG were isolated from 8 - to 10-week-old animals and immediately fixed in 4% paraformaldehyde (Sigma) rinsed in PBS (0.01 M, pH 7.4) at 4℃ for three days. After the fixation, we replaced the fixative solution with cryoprotective solution of 30% sucrose (Sigma). The spinal cord and DRG were cut into 10 and 20 µm sections, respectively. The 20 µm sections were incubated with hematoxylin and eosin staining solutions, and the 10 µm were incubated with rabbit anti-TRPV1 (1:50, Santa Cruz) and mouse anti-NeuN (1:2000, Millipore) overnight at 4℃ in 1% BSA with 0.05% of Triton, then incubated with Cy2-donkey anti-mouse and Cy3-donkey anti-rabbit (1:200, from Jackson ImmunoResearch) for 2 h at Troom in dark. DAPI (dilactate; 300 nM, Invitrogen) was used for counterstaining. Images were taken by a confocal microscope.
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