EnVision (EnVision + , Dako, Carpentaria, CA) system was used for immunohistochemical detection of human and mouse skin tissues. In brief, the skin tissue samples were shaved and rehydrated in double distilled water. Next, 3% H2O2 was used to block endogenous peroxidase activity, and then sodium citrate solution was used for antigen repair. Then, antibodies to IL-32 (ab37158, 1: 100, Abcam), JAK1 (ab125051, 1: 100, Abcam), p-JAK1 (PA5-104554, 1: 100, Invitrogen), IL-32 (ab37158, 1: 100, Abcam), p-STAT1 (#9167, 1: 800, Cell Signaling Technology), STAT1 (ab230428, 1: 100, Abcam), STAT3 (ab68153, 1: 100, Abcam), and p-STAT3 (ab76315, 1: 100, Abcam) were used to incubate the samples at 37 °C for 3 h. Then, the sample was further reacted with secondary antibody obtained from the Zhongshan Biotechnology company (Beijing, China). After diaminobenzidine staining, routine staining was performed followed by microscopic examination.
Ab125051
Manufactured by Abcam
Sourced in China
Ab125051 is a lab equipment product manufactured by Abcam. It is a tool used for scientific research and analysis, but a detailed description of its core function cannot be provided in a concise, unbiased, and factual manner without extrapolation. Therefore, the description is not available.
Automatically generated - may contain errors
Lab products found in correlation
Ab68153, by Abcam (2 mentions)
Ab230428, by Abcam (1 mentions)
Ab37158, by Abcam (1 mentions)
Ab76315, by Abcam (1 mentions)
Ab150078, by Abcam (1 mentions)
Ab104139, by Abcam (1 mentions)
Ab5690, by Abcam (1 mentions)
Enhanced chemiluminescence western blotting substrate, by GE Healthcare (1 mentions)
Ab32520, by Abcam (1 mentions)
Ab16276, by Abcam (1 mentions)
Anti jak1 rabbit monoclonal antibody, by Cell Signaling Technology (1 mentions)
Horseradish peroxidase conjugated goat anti rabbit igg, by Agilent Technologies (1 mentions)
Anti halotag monoclonal antibody, by Promega (1 mentions)
Ab9485, by Abcam (1 mentions)
4 protocols using ab125051
1
Immunohistochemical Analysis of Skin Tissues
2
Immunofluorescence Staining of JAK1 in Cells
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The cell culture medium was aspirated, and the cells were washed twice with PBST. The cells were then fixed in a 4 % paraformaldehyde solution for 20 min, followed by three PBST washes to remove any residual fixative. The cells were blocked and permeabilized with a solution of PBS with 5 % normal goat serum and 0.1 % Triton X-100 for 2 h at room temperature. Subsequently, SCs were incubated overnight at 4 °C with a primary antibody, Rabbit polyclonal to JAK1 (5 μg/ml, ab125051, Abcam), diluted in the blocking buffer. Afterwards, cells were rinsed twice with PBS and then washed three times with PBST. Next, primary antibody labled cells were incubated with a goat anti-rabbit IgG H&L secondary antibody (Alexa Fluor® 555) (1:500, ab150078, Abcam) for 2 h at room temperature while avoiding light. Following another round of PBS washing, the samples were mounted with Fluoroshield Mounting Medium with DAPI (ab104139, Abcam). Finally, the samples were observed and photographed using a confocal microscope.
3
Oxymatrine Modulation of Inflammatory Cytokines
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Oxymatrine (Y30S6-Y17043, HPLC grade, purity ≥ 98%) was purchased from Shanghai Yuanye Bio-Technology Co., Ltd. Interleukin-4 (IL-4) ELISA kit (EK0405), interleukin-6 (IL-6) ELISA kit (EK0411), interleukin-17 (IL-17) ELISA kit (EK0431) and tumor necrosis factor-α (TNF-α) ELISA kit (EK0527) were purchased from Wuhan Boster Biological Engineering Co., Ltd. Immunoglobulin E (IgE) ELISA kit (bsk12030) was purchased from Beijing Bioss Biotechnology Co., Ltd. Rabbit anti-mouse antibodies such as SOCS1 (ab280886), T lymphocyte surface marker CD3 (ab5690), JAK1 (ab125051), STAT3 (ab68153), and phosphorylated JAK1 (p-JAK1) (ab1380056) were purchased from the Abcam Co., Ltd.
4
Detecting JAK/STAT Pathway Activation
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Zirconia balls were added to the proteins extracted from the liver of P0 newborns. Then, the proteins were dissolved in 1 ml sample buffer (NuPAGE LDS sample buffer 250μL, sample reducing agent 100μL, 25×protease inhibiter 40μL, and water 610μL) and crushed. After centrifugation at 10,000 rpm for 10 min at 4°C, the supernatant of each sample was subjected to SDS-PAGE. Strips of membrane were incubated with anti-p-JAK1 (Tyr1021), anti-JAK1 (ab125051; Abcam), anti-p-STAT1 (Tyr701), anti-STAT1 (ab99415; Abcam), anti-p-STAT3 (Tyr705), anti-STAT3 (SAB4300708, Sigma Aldrich), anti-p-STAT5 (Tyr694), anti-STAT5 (ab16276; Abcam), anti-p-STAT6 (Tyr641), anti-STAT6 (ab32520; Abcam) or anti-GAPDH (ab9485; Abcam) antibodies. The antibody–antigen complexes were detected with horseradish peroxidase–conjugated goat anti-rabbit IgG (Dako, Glostrup, Denmark) at a dilution of 1:1,000, followed by detection with enhanced chemiluminescence Western blotting substrate (GE Healthcare BioSciences, Little Chalfont, UK), as described by the manufacturer. n=3. Each experiment was performed three times. For HEK293 cell lysates, additional anti-HaloTag monoclonal antibody (G9211; Promega Corporation, WI) and anti-JAK1 rabbit monoclonal antibody (#3344; Cell Signaling Technology, MA) were used as the primary antibodies.
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