Collagenase type I, DNase I, and Pronase were purchased from Sigma Aldrich. The anti-CD133 antibody was purchased from Miltenyi Biotech. The anti-αSMA, anti-IL6, anti-pNFκB (S536), anti-NFκB, and anti-TAK1 antibodies were purchased from Abcam. The anti-β2SP antibody was a gift from Dr. Lopa Mishra's laboratory. The anti-pYSTAT3, anti-STAT3, anti-TAK1, anti-pIκK α/β and anti-pTAK1 antibodies were purchased from Cell Signaling Technology. The anti-IκKβ was purchased from R&D systems.
Anti tak1
Manufactured by Abcam
Sourced in United Kingdom
Anti-TAK1 is a monoclonal antibody that specifically recognizes the TAK1 protein. TAK1 is a serine/threonine protein kinase that plays a role in various cellular signaling pathways.
Automatically generated - may contain errors
Lab products found in correlation
Sybr green master mix, by Vazyme (2 mentions)
Phenylmethanesulfonyl uoride pmsf, by Thermo Fisher Scientific (2 mentions)
Anti p38, by Abcam (2 mentions)
Anti p p38, by Abcam (2 mentions)
Anti erk1 2, by Abcam (2 mentions)
Anti jnk, by Abcam (2 mentions)
Anti p jnk, by Abcam (2 mentions)
Anti p nf κb, by Abcam (2 mentions)
Anti p tak1, by Abcam (2 mentions)
Anti nf κb, by Abcam (2 mentions)
Sb202190, by MedChemExpress (2 mentions)
Ros assay kit, by Beyotime (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Trizol, by Thermo Fisher Scientific (2 mentions)
Dulbecco s modi ed eagle s medium dmem, by Thermo Fisher Scientific (2 mentions)
Radioimmunoprecipitation assay (ripa), by Beyotime (2 mentions)
Hiscript reverse transcriptase, by Vazyme (2 mentions)
Bca protein assay kit, by Beyotime (2 mentions)
Phosphatase inhibitor cocktail, by Beyotime (2 mentions)
Anti p erk1 2, by Abcam (2 mentions)
6 protocols using anti tak1
1
Cell Isolation and Characterization Protocol
2
Western Blot Analysis of Signaling Proteins
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Whole-cell protein extracts were resolved by SDS-polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride (PVDF; Millipore, Billerica, MA). Blots were probed with anti-NFX1-123 (1:1000), anti-p53 (1:1000, Calbiochem, San Diego, CA), (anti-TRAF6 (1:500; Cell Signaling, Danvers, MA), anti-TRIF (1:1000, Abcam, Cambridge, MA), anti-TAK1 (1:500, Abcam, Cambridge, MA), anti-TAB1 (1:500, Abcam, Cambridge, MA), anti-TAB2 (1:500, Abcam, Cambridge, MA), and anti-GAPDH (1:100,000; Abcam, Cambridge, MA) primary antibodies. The secondary antibodies used were anti-mouse IgG HRP (1:10,000; Cell Signaling, Danvers, MA) or anti-rabbit IgG HRP (1:5,000, Cell Signaling, Danvers, MA). The rabbit polyclonal anti-NFX1-123 antibody was generously provided by Dr. Ann Roman. Animals were immunized with peptide composed of amino acids 1102–1120 of NFX1-123. All films were scanned using Epson Perfection V700 and imported using Adobe Photoshop.
3
Inflammasome Activation Pathway Assay
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Anti-IL-1β, Anti-Caspase1, TNF-α were purchased from R&D Systems (Minneapolis, MN, USA); Anti-cleaved caspase1, Anti-NLRP3, Anti-IκBα, Anti-phospho-P38, Anti-phospho-ERK, Anti-phospho-JNK, Anti-phospho-IKKα/β, and Anti-Tublin were purchased from Cell Signaling Technology (Boston, MA, USA); Anti-TAK1, Anti-phospho-TAK1, Anti-ASC and recombinant active caspase1 protein were purchased from Abcam (Shanghai, China). The ultrapure LPS and Nigericin were obtained from Sigma-Aldrich (St. Louis, MO, USA). The caspase1 activity assay kit was purchased from Beyotime Biotechnology (Shanghai, China). The Human IL-1β ELISA Kit was purchased from Boster Biological Technology (Wuhan, China).
4
Western Blot Analysis of Inflammatory Signaling
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Whole protein in the serum was collected using a whole-protein extraction kit (Nanjing KeyGEN Biotech, Nanjing, China) according to the manufacturer’s instructions. The protein concentration was measured using a bicinchoninic acid kit (Nanjing KeyGEN Biotech). The different proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and transferred onto polyvinylidene difluoride membranes (Bio-Rad). The membranes were blocked in Tris-buffered saline and Tween 20 (TBST) blocking buffer that contained 5% nonfat dried milk for 2 h at room temperature and immunoblotted with primary specific antibodies (anti-TLR4, anti-TRAF6, anti-TAK1, IκB-α, p-IκB-α, anti-NF-κB (p65), anti-p-NF-κB (p-p65), and anti-β-actin (Abcam, UK). After washing with TBST, the strips were incubated with horse-radish peroxidase-conjugated goat anti-mouse IgG (HuaAn, Hangzhou, China). Proteins were detected using a SuperSignal™ West Femto Maximum Sensitivity Substrate (Thermofisher) and visualized using a ChemiDoc™ MP Imaging System (Bio-Rad).
5
Inflammatory Signaling Pathways in RAW264.7 Cells
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Materials BAC (purity ≥ 98%) and RAW264.7 cells were provided by Yousi Scienti c Co., Ltd., and the Chinese Academy of Sciences (Shanghai, China). LPS from Escherichia coli and TPCK (NF-κB inhibitor) were obtained from Sigma (St. Louis, MO, USA). SB202190 (p38 MAPK inhibitor), U0126 (ERK inhibitor), SP600125 (JNK inhibitor), and Cell-Counting Kit-8 (CCK-8) were obtained from Medchem Express (St.
Louis, MO, USA). Trizol and phenylmethanesulfonyl uoride (PMSF) were acquired from Ambion (CA, USA) and Aladdin (Shanghai, China). HiScript Reverse Transcriptase and SYBR Green Master Mix were provided by VAZYME (Nanjing, China). ROS Assay Kit, BCA Protein Assay Kit, radioimmunoprecipitation assay (RIPA), and phosphatase inhibitor cocktail were acquired from Beyotime (Shanghai, China).
Dulbecco's modi ed Eagle's medium (DMEM) and fetal bovine serum (FBS) were obtained from Gibco (Grand Island, NY). Anti-NF-κB, anti-JNK, anti-p38, anti-ERK1/2, anti-TAK1, anti-p-NF-κB, anti-p-JNK, anti-p-p38, anti-p-ERK1/2, and anti-p-TAK1 antibodies were purchased from Abcam (Cambridge, UK). Anti-IκBα and anti-p-IκBα antibodies were obtained from A nity Bio Reagents (Golden, CO, USA).
Louis, MO, USA). Trizol and phenylmethanesulfonyl uoride (PMSF) were acquired from Ambion (CA, USA) and Aladdin (Shanghai, China). HiScript Reverse Transcriptase and SYBR Green Master Mix were provided by VAZYME (Nanjing, China). ROS Assay Kit, BCA Protein Assay Kit, radioimmunoprecipitation assay (RIPA), and phosphatase inhibitor cocktail were acquired from Beyotime (Shanghai, China).
Dulbecco's modi ed Eagle's medium (DMEM) and fetal bovine serum (FBS) were obtained from Gibco (Grand Island, NY). Anti-NF-κB, anti-JNK, anti-p38, anti-ERK1/2, anti-TAK1, anti-p-NF-κB, anti-p-JNK, anti-p-p38, anti-p-ERK1/2, and anti-p-TAK1 antibodies were purchased from Abcam (Cambridge, UK). Anti-IκBα and anti-p-IκBα antibodies were obtained from A nity Bio Reagents (Golden, CO, USA).
6
Inflammatory Signaling Pathways in RAW264.7 Cells
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Materials BAC (purity ≥ 98%) and RAW264.7 cells were provided by Yousi Scienti c Co., Ltd., and the Chinese Academy of Sciences (Shanghai, China). LPS from Escherichia coli and TPCK (NF-κB inhibitor) were obtained from Sigma (St. Louis, MO, USA). SB202190 (p38 MAPK inhibitor), U0126 (ERK inhibitor), SP600125 (JNK inhibitor), and Cell-Counting Kit-8 (CCK-8) were obtained from Medchem Express (St.
Louis, MO, USA). Trizol and phenylmethanesulfonyl uoride (PMSF) were acquired from Ambion (CA, USA) and Aladdin (Shanghai, China). HiScript Reverse Transcriptase and SYBR Green Master Mix were provided by VAZYME (Nanjing, China). ROS Assay Kit, BCA Protein Assay Kit, radioimmunoprecipitation assay (RIPA), and phosphatase inhibitor cocktail were acquired from Beyotime (Shanghai, China).
Dulbecco's modi ed Eagle's medium (DMEM) and fetal bovine serum (FBS) were obtained from Gibco (Grand Island, NY). Anti-NF-κB, anti-JNK, anti-p38, anti-ERK1/2, anti-TAK1, anti-p-NF-κB, anti-p-JNK, anti-p-p38, anti-p-ERK1/2, and anti-p-TAK1 antibodies were purchased from Abcam (Cambridge, UK). Anti-IκBα and anti-p-IκBα antibodies were obtained from A nity Bio Reagents (Golden, CO, USA).
Louis, MO, USA). Trizol and phenylmethanesulfonyl uoride (PMSF) were acquired from Ambion (CA, USA) and Aladdin (Shanghai, China). HiScript Reverse Transcriptase and SYBR Green Master Mix were provided by VAZYME (Nanjing, China). ROS Assay Kit, BCA Protein Assay Kit, radioimmunoprecipitation assay (RIPA), and phosphatase inhibitor cocktail were acquired from Beyotime (Shanghai, China).
Dulbecco's modi ed Eagle's medium (DMEM) and fetal bovine serum (FBS) were obtained from Gibco (Grand Island, NY). Anti-NF-κB, anti-JNK, anti-p38, anti-ERK1/2, anti-TAK1, anti-p-NF-κB, anti-p-JNK, anti-p-p38, anti-p-ERK1/2, and anti-p-TAK1 antibodies were purchased from Abcam (Cambridge, UK). Anti-IκBα and anti-p-IκBα antibodies were obtained from A nity Bio Reagents (Golden, CO, USA).
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