Isolated proteins (30 µg from every specimen as evaluated via BCA protein assay) were subjected to electrophoresis (8%‒12 % SDS-Tris glycine gel) and were transferred to the PVDF membranes. Membranes were blocked with the help of 5% skimmed milk within TBS-T buffer (10 mM Tris [pH 7.5], 100 mM NaCl, 0.1% Tween 20) and incubated with primary antibodies (GAPDH antibody [1:5000, ab8245, Abcam], IL-1beta antibody [1:5000, ab9722, Abcam], IL-6 antibody [1:2500, ab6672, Abcam], TNF-alpha antibody [1:2500, ab6671, Abcam], TRPV1 antibody [1:1000, ab10296, Abcam], phosphor-AKT antibody [1:1000, ab66138, Abcam], AKT antibody [1:5000, ab8805, Abcam], PI3K antibody [1:2000, ab86714, Abcam], phosphor-p65 antibody [1:1000, ab86299, Abcam], and p65 antibody [1:5000, ab16502, Abcam]) at room temperature. The membranes were then incubated with secondary antibodies, which were anti-rabbit antibodies (1:5000) conjugated with peroxidase. Bands were observed via an enhanced chemiluminescent substrate kit (provided by PIERCE) with the help of LAS-3000 Fujiflm (Fuji Photo Film Co. Ltd). Band intensity was quantified using ImageJ software (Bethesda, MD, USA).
Ab10296
Manufactured by Abcam
Sourced in United States
Ab10296 is a rabbit monoclonal antibody that specifically recognizes the target protein. It is suitable for use in various immunoassay applications.
Automatically generated - may contain errors
Lab products found in correlation
Ab6672, by Abcam (1 mentions)
Ab6671, by Abcam (1 mentions)
Ab8805, by Abcam (1 mentions)
Ab86714, by Abcam (1 mentions)
Ab66138, by Abcam (1 mentions)
Ab16502, by Abcam (1 mentions)
Ab8245, by Abcam (1 mentions)
Ab9722, by Abcam (1 mentions)
Ab86299, by Abcam (1 mentions)
Ab6199, by Abcam (1 mentions)
Ab124760, by Abcam (1 mentions)
Ab15191, by Abcam (1 mentions)
Ab46154, by Abcam (1 mentions)
Gk346810, by Leica (1 mentions)
Ab34710, by Abcam (1 mentions)
Goat anti mouse igg hrp, by Santa Cruz Biotechnology (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Capsaicin, by Merck Group (1 mentions)
Ab5694, by Abcam (1 mentions)
Tgf β, by R&D Systems (1 mentions)
4 protocols using ab10296
1
Western Blot Analysis of Inflammatory Markers
2
Immunohistochemical Analysis of Endometrial Cysts
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Tissues blocks were prepared and sectioned at 7 μm, deparaffinized, and rehydrated. Sections from endometrial cysts were then incubated overnight with anti-NGF antibody (1:200, ab6199; Abcam, Cambridge, Britain), anti-COX2 antibody (1:600, ab15191; Abcam), anti-ICAM antibody (1:1000, ab124760; Abcam), and anti-VEGF antibody (1:200, ab46154; Abcam). Sections from DRG were incubated with anti-NGF antibody (1:200, ab6199; Abcam) and anti-TRPV1 antibody (1:1000, ab10296; Abcam). After washing with 1× PBS, the sections were incubated with secondary antibody. Envision-labeled polymer-alkaline phosphatase rabbit (GK400305/15, Novocastra, UK) and diaminobenzidine (GK346810, Novocastra, UK) were used to visualize the antigen–antibody reaction. The immunohistochemistry (IHC) score was the sum of percentage (0–3) and intensity scores (0–3). Using a Zeiss microscope, the average IHC score of five different fields at 400x magnification was regarded as the molecule expression. All slides were analyzed by two blinded observers.
3
Evaluating Fibrosis Markers in CCl4-Induced Liver Injury
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CCl4 was obtained from the Shantou Xilong Chemistry Plant (Shantou, Guangdong, China). Dimethylsulfoxide (DMSO) was purchased from Sigma-Aldrich. (St. Louis, MO, USA). Antibodies against TRPV1 (ab10296), α-SMA (ab5694), COL1A1 (ab34710), and β-actin (ab8226) were obtained from Abcam (Cambridge, MA, USA), same as the picro-sirius red staining kit (ab150681). TGF-β was purchased from R&D Systems (Minneapolis, MN, USA). Secondary antibodies for goat anti-rabbit immunoglobulin (IgG), horse radish peroxidase (HRP), and goat anti-mouse IgG HRP were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Capsaicin was purchased from Sigma (Santa Clara, CA, USA).
4
5-HT2B Receptor Agonist and Antagonist Protocol
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The 5-HT2B receptor agonist (BW723C86; Sigma–Aldrich, St. Louis, MO, United States) was diluted with saline. The 5-HT2B receptor antagonist 2-amino-4-(4-fluoronaphth-1-yl)-6-isopropylpyridine (RS-127445; Tocris Bioscience, Ellisville, Missouri, United States) was dissolved in 20% dimethylsulfoxide in physiological saline. The TRPV1 antagonist N-(3-methoxyphenyl)-4-chlorocinnamide (SB366791; Sigma–Aldrich, St. Louis, MO, United States) was dissolved in 50% dimethylsulfoxide in physiological saline. A human anti-5-HT2B receptor polyclonal antibody (MAB10322) was purchased from R&D Systems (Minneapolis, MN, United States). Mouse anti-5-HT2B receptor (AB194333), anti-alpha smooth muscle actin (AB5694) and rabbit anti-TRPV1 polyclonal antibodies (AB10296) were purchased from Abcam (Cambridge, MA, United States). Rabbit anti-Ecadherin antibody (CST 3195) was purchased from Cell Biological Technology, Inc. (Boston, United States). The rabbit anti-protein-encoding gene product (PGP) 9.5 (14730-1-AP) polyclonal antibody, rabbit anti-pankeratin (26411-1-AP) polyclonal antibody, and mouse anti-TRPV1 polyclonal antibody (66983-1-Ig) were obtained from Proteintech Group, Inc. (Chicago, United States). A 5-HT2B receptor ELISA kit was purchased from Jianglai Bio (Shanghai, China). A RNA polymerase chain reaction (PCR) kit was purchased from Takara Biotechnology (Dalian, China).
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