Western blot analysis was performed to determine STAT3 expression in vitro. HaCaT cells were treated with PBS, Dextran (20 μg/ml), ACT (20 μg/ml)or ACT-Dex NPs (20 µg ACT equiv./mL) for 12 h. Subsequently, the cells were further stimulated using recombinant IL-22 (PeproTech; 50 ng/ml) for an additional 1 h, and the cell lysate was prepared. After electrophoresis, proteins were electroeluted onto a polyvinylidenedifluoride (PVDF) membrane (Invitrogen, Carlsbad, USA). Antibodies: anti-pSTAT3 (Tyr705) (Cell Signaling, Beverly, USA), STAT3 (Cell Signaling), and anti-actin (Abcam, Cambridge, United Kingdom). For the visualization of the immunoreactive proteins, an enhanced chemiluminescence assay kit and a chemiluminescence imaging system (ChemiScope 6200T, ClinX, Shanghai, China) were used.
Chemiscope 6200t
Manufactured by Clinx
Sourced in China
The ChemiScope 6200T is a high-precision laboratory instrument designed for chemical analysis and research. It provides accurate and reliable measurements for a variety of applications. The core function of the ChemiScope 6200T is to perform advanced spectroscopic analysis using state-of-the-art technology.
Automatically generated - may contain errors
Lab products found in correlation
Ab2787, by Abcam (2 mentions)
Ab125011, by Abcam (2 mentions)
Ab92726, by Abcam (2 mentions)
Ab8226, by Abcam (2 mentions)
Ba1055, by Boster Bio (2 mentions)
Radioimmunoprecipitation assay buffer, by Beyotime (2 mentions)
Anti pstat3 tyr705, by Cell Signaling Technology (1 mentions)
Recombinant il 22, by Thermo Fisher Scientific (1 mentions)
Anti actin, by Abcam (1 mentions)
Stat3, by Cell Signaling Technology (1 mentions)
Polyvinylidene difluoride pvdf membrane, by Thermo Fisher Scientific (1 mentions)
Ab40772, by Abcam (1 mentions)
Ab179800, by Abcam (1 mentions)
Ab7817, by Abcam (1 mentions)
S0009, by Affinity Biosciences (1 mentions)
Ab92547, by Abcam (1 mentions)
Ab203676, by Abcam (1 mentions)
Ab270993, by Abcam (1 mentions)
Ab264077, by Abcam (1 mentions)
4 protocols using chemiscope 6200t
1
STAT3 Expression Analysis in HaCaT Cells
2
Protein Expression Analysis of HK-2 Cells
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HK-2 cells were lysed in radioimmunoprecipitation assay buffer (Beyotime, China). Immunoblotting was performed using the following rabbit primary antibodies: anti-CD9 (1:2,000; ab92726, Abcam, UK), anti-TSG101 (1:2,000; ab125011, Abcam), anti-HSP70 (1:1,000; ab2787, Abcam), anti-vimentin (1:1,000; ab92547, Abcam), anti-α-smooth muscle actin (anti-α-SMA; 1:1,000; ab7817, Abcam), anti-E-cadherin (anti-ECAD; 1:10,000; ab40772, Abcam), anti-cyclooxygenase 2 (anti-COX-2; 1:1,000; ab179800, Abcam), anti-AKT [1:1,000; 4691S, Cell Signaling Technology (CST), USA], anti-p-AKT (1:1,000; 4060S, CST), anti-mitogen-activated protein kinase (MAPK) (ERK1/2) (1:1,000; 8544S, CST), anti-p-MAPK (ERK1/2) (1:1,000; 8544S, CST), anti-p38 (1:1,000; 8690S, CST), anti-p-p38 (1:1,000; 9216S, CST), anti-SOX2 (1:1,000; AF5140, Affinity, Canada), and anti-β-actin (1:5,000; ab8226, Abcam). Horseradish-peroxidase-conjugated goat anti-rabbit immunoglobulin G (IgG) (1:5,000; BA1055, Boster, China) and horseradish-peroxidase-conjugated goat anti-rat IgG (1:5,000; S0009, Affinity, Canada) were used as the secondary antibodies. A chemiluminescence imaging system (ChemiScope 6200T, Clinx Science Instruments, Shanghai, China) was used for detection.
3
Immunoblotting Analysis of AIF Proteins
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Hippocampal tissue was collected in an immunoprecipitation buffer and analyzed by immunoblotting with an AIF mouse or rabbit anti‐Flag antibody. Proteins were separated using SDS‐PAGE and transferred to a PVDF membrane. The membrane was then blocked and incubated overnight with a 1 mg/mL rabbit anti‐Flag (anti‐SV2A, anti‐MIF, anti‐H2AX) or a mouse anti‐PAR primary antibody at 4°C, followed by a goat anti‐rabbit IgG or a donkey anti‐mouse IgG conjugated to HRP for 1 h at room temperature. After washing, immune complexes were detected using the Chemi‐luminescent system (Chemi‐scope 6200(T), CLINX, Shanghai, China).
4
Characterization of Tendon Cell Markers
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TSCs were lysed in radioimmunoprecipitation assay buffer (Beyotime). Immunoblotting was performed using the following rabbit primary antibodies: anti-CD9 (monoclonal; 1:2000; ab92726; Abcam, Cambridge, UK), abti-TSG101 (monoclonal; 1:2000; ab125011; Abcam), anti-Hsp70 (monoclonal; 1:1000; ab2787; Abcam), anti-tenomodulin (anti-TNMD; polyclonal; 1:1000; ab203676; Abcam), anti-collagen I (monoclonal; 1:1000; ab270993; Abcam), anti-scleraxis (anti-SCXA; polyclonal; 1:500; DF13293; Affinity Biologicals, Ancaster, ON, Canada), anti-alkaline phosphatase (anti-ALP; polyclonal; 1:1000; DF6225; Affinity Biologicals), anti-runt-related transcription factor 2 (anti-Runx2; monoclonal; 1:1000; ab264077; Abcam), anti-SMAD2/3 (monoclonal; 1:1000; 5678S; Cell Signaling Technology, Danvers, MA, USA), anti-phospho (p)-SMAD2/3 (monoclonal; 1:1000; 8828S; Cell Signaling Technology), anti-SMAD1/5/9 (polyclonal; 1:500; AF0614; Affinity Biologicals), anti-phospho-SMAD1/5/9 (polyclonal; 1:1000; AF8313; Affinity Biologicals), and anti-β-actin (monoclonal; 1:5000; ab8226; Abcam). Horseradish peroxidase-conjugated goat anti-rabbit IgG (1:5000; BA1055; Boster, Wuhan, China) was used as the secondary antibody. A chemiluminescence imaging system (ChemiScope 6200T, Clinx Science Instruments, Shanghai, China) was used for detection.
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