Each tissue slide was prepared as described previously [69 (link)]. Staining was carried out via H&E and toluidine blue, followed by immunohistochemical staining with anti-filaggrin (Abcam, Cambridge, UK), anti-loricrin (Abcam), and anti-TRPA1 (GTX54765; GeneTex, Irvine, CA, USA) antibodies. Staining was performed using an Ultravision Quanto Detection System (TL-060-QHD; Thermo Fisher Scientific, Waltham, MA, USA). After staining, tissues were dehydrated and sealed in Permount (SP15-100; Thermo Fisher Scientific), and observed via optical microscopy (DM750, Leica, Wetzlar, Germany). For fluorescence images, the sections were incubated with primary antibody with anti-TRPA1 (GTX54765; GeneTex, Irvine, CA, USA) antibody at 4 °C for 16 h, followed by incubation with secondary antibodies against FITC-conjugated goat-anti-rabbit IgG (1:1000, sc-2012, Santa Cruz Biotechnology) at 21 °C for 1 h. Immunostained tissues were mounted with a medium containing Fluoroshield™ with DAPI (ImmunoBioScience, Mukilteo, WA, USA). Fluorescence images were acquired by using a confocal microscope (LSM700; Zeiss, Jena, Germany).
Anti trpa1 gtx54765
Manufactured by GeneTex
Sourced in United States
Anti-TRPA1 (GTX54765) is a laboratory reagent produced by GeneTex. It is an antibody that specifically targets the TRPA1 protein. TRPA1 is a member of the transient receptor potential (TRP) ion channel family and is involved in various physiological and pathological processes. The core function of this product is to enable the detection and analysis of TRPA1 in research applications.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 700, by Zeiss (2 mentions)
Fluorescence microscope, by Leica (1 mentions)
Dm750, by Leica (1 mentions)
Permount sp15 100, by Thermo Fisher Scientific (1 mentions)
Ultravision quanto detection system tl 060 qhd, by Thermo Fisher Scientific (1 mentions)
Anti filaggrin, by Abcam (1 mentions)
Anti loricrin, by Abcam (1 mentions)
Fitc conjugated goat anti rabbit igg, by Santa Cruz Biotechnology (1 mentions)
2 protocols using anti trpa1 gtx54765
1
Immunohistochemical Analysis of Skin Samples
2
Immunohistochemical Analysis of DRG Nociceptors
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Rats were terminally anesthetized with sevoflurane and perfused transcardially
with PBS followed by 4% paraformaldehyde. After perfusion, the L3-5 DRGs were
collected, post-fixed in 4% paraformaldehyde overnight at 4°C, and then
dehydrated in sucrose. Later, tissues were embedded in OCT (Tissue-Tek, Japan)
and serially sectioned in a cryostat into 15-μm-thick slices. The tissue
sections were blocked with 10% goat serum in 0.3% Triton for 1 h at room
temperature and incubated primary antibodies (anti-TRPA1, GTX54765, GeneTex,
USA; anti-TLR4, GTX57153, GeneTex, USA) overnight in a wet box at 4°C.
Subsequently, the sections were incubated with corresponding secondary
antibodies for 1 h at room temperature. The stained sections were visualized
under a fluorescence microscope (Leica Microsystems, Wetzlar, Germany). Three
DRG tissues (L3-L5) in each tested rat were taken for IF staining and three
sections were randomly selected in each tested DRG. Moreover, three rats were
randomly selected in each group for IF evaluation. The images for each target
were captured under same exposure time and fluorescence intensity in each image
was quantized by Image J software for data analysis.
with PBS followed by 4% paraformaldehyde. After perfusion, the L3-5 DRGs were
collected, post-fixed in 4% paraformaldehyde overnight at 4°C, and then
dehydrated in sucrose. Later, tissues were embedded in OCT (Tissue-Tek, Japan)
and serially sectioned in a cryostat into 15-μm-thick slices. The tissue
sections were blocked with 10% goat serum in 0.3% Triton for 1 h at room
temperature and incubated primary antibodies (anti-TRPA1, GTX54765, GeneTex,
USA; anti-TLR4, GTX57153, GeneTex, USA) overnight in a wet box at 4°C.
Subsequently, the sections were incubated with corresponding secondary
antibodies for 1 h at room temperature. The stained sections were visualized
under a fluorescence microscope (Leica Microsystems, Wetzlar, Germany). Three
DRG tissues (L3-L5) in each tested rat were taken for IF staining and three
sections were randomly selected in each tested DRG. Moreover, three rats were
randomly selected in each group for IF evaluation. The images for each target
were captured under same exposure time and fluorescence intensity in each image
was quantized by Image J software for data analysis.
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