βBA, obtained from Sigma-Aldrich (St. Louis, MO, USA), was dissolved and prepared with 5, 10, 20, and 30 mM stock solutions in DMSO. The control group was added 0.1% DMSO. TRIzol reagent was obtained from Life Technologies (Carlsbad, CA, USA). A monoclonal anti-β-actin antibody was obtained from Sigma-Aldrich (St. Louis, MO, USA). Antibodies against phospho-Akt, anti-Akt, anti-phospho-p38, anti-total p38, anti-phospho-IκB, anti-Bruton’s tyrosine kinase, and anti-phospho-PLCγ2 were obtained from Cell Signaling Technology Inc. (Beverly, MA, USA). Anti-phospho-Btk antibody was obtained from GeneTex (Irvine, CA, USA). Anti-c-Fos, anti-NFATc1, anti-IκB, and anti-PLCγ2 antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Donkey anti-rabbit and anti-mouse immunoglobulin secondary antibodies were purchased from Enzo Life Sciences (Farmingdale, NY, USA).
Anti total p38
Manufactured by Cell Signaling Technology
Sourced in United States
Anti-total p38 is a laboratory reagent used to detect the p38 protein in a variety of samples. It is a primary antibody that binds to the p38 protein, allowing for its identification and quantification through various experimental techniques.
Automatically generated - may contain errors
Lab products found in correlation
Anti phospho p38, by Cell Signaling Technology (9 mentions)
Anti total jnk, by Cell Signaling Technology (7 mentions)
Anti phospho jnk, by Cell Signaling Technology (6 mentions)
Anti c fos, by Santa Cruz Biotechnology (3 mentions)
β actin, by Merck Group (3 mentions)
Anti total erk, by Cell Signaling Technology (3 mentions)
Anti phospho erk, by Cell Signaling Technology (3 mentions)
Anti phospho akt, by Cell Signaling Technology (3 mentions)
Anti total erk1 2, by Cell Signaling Technology (3 mentions)
Anti akt, by Cell Signaling Technology (2 mentions)
Donkey anti rabbit and anti mouse immunoglobulin secondary antibodies, by Enzo Life Sciences (2 mentions)
Trizol reagent, by Thermo Fisher Scientific (2 mentions)
Anti plcγ2, by Santa Cruz Biotechnology (2 mentions)
Anti iκb, by Santa Cruz Biotechnology (2 mentions)
Anti phospho iκb, by Cell Signaling Technology (2 mentions)
Anti nfatc1, by Santa Cruz Biotechnology (2 mentions)
Anti bcl 2, by Cell Signaling Technology (2 mentions)
Anti bax, by Cell Signaling Technology (2 mentions)
Heme oxygenase 1 ho 1, by Abcam (2 mentions)
Anti total akt, by Cell Signaling Technology (2 mentions)
21 protocols using anti total p38
1
Investigating β-Actin Regulation Pathways
2
Angiotensin Receptor Signaling Pathway
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Angiotensin converting enzyme (ACE), and angiotensin II type 1 receptor (AT-1R) (Santa Cruz Biotechnology, Santa Cruz, CA, USA), ACE2 (R&D, Minneapolis, MN, USA), TACE (Millipore), heme oxygenase-1 (HO-1, Abcam, Inc., Cambridge, MA, USA), angiotensin II/III (Novus Biologicals, Littleton CO, USA), anti-p-p38, anti-total p38, anti-c-Jun N-terminal kinase (JNK), anti-p-JNK, anti-caspase-3, anti-cleaved caspase-3, (Cell Signaling Technology, Danvers, MA), anti-Bcl-2, and anti-Bax (Cell Signaling Technology, Beverly, MA, USA), and β-actin (Sigma-Aldrich, St. Louis, MO, USA) antibodies were purchased.
3
Regulation of T-cell Signaling Pathways
4
Western Blot Analysis of Cardiac Proteins
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Western blots to analyse protein expression were conducted using the standard protocol as described previously 17 , 18 . Primary antibodies used were anti‐L‐type Ca2+ channel (Abcam, Cambridge, UK), anti‐ryanodine receptor (Abcam), anti‐SERCA2a (Santa Cruz Biotechnology, Dallas, TX, USA), anti‐NCX (Cell Signaling, Danvers, MA, USA), anti‐phospho‐PLB (Millipore, Bellerica, MA, USA), anti‐total‐PLB (Millipore), anti‐PMCA1 (Abcam), anti‐PMCA4 (Abcam), anti‐RAGE (Abcam), anti‐phospho‐p38 (Cell Signaling), anti‐total‐p38 (Cell Signaling), anti‐iNOS (Abcam), anti‐α‐tubulin (Calbiochem) and anti‐GAPDH (Santa Cruz). For visualization, we used HRP‐conjugated secondary antibodies, which were obtained from either Cell Signaling or Dako.
5
Duoxa1 Overexpression in Mouse Cells
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Full-length wild-type Duoxa1 was amplified using PCR from mouse cDNA and ligated into the pMX-IRES-EGFP vector as pMX-Duoxa1 using the BamHI and XhoI (Enzynomics, Daejeon, Korea) sites. The following primers were used: Duoxa1-For, 5′-GCTAGGATCCATGGCTGCTCTTGGACACAC-3′ and Duoxa1-Rev, 5′-CGACTCGAGCAGGGAACAGTCGGACTCTTTG-3′. TRIzol reagent was obtained from Life Technologies (Carlsbad, CA, USA). A monoclonal β-actin (A5441) antibody and DAPI (D9542) were obtained from Sigma (St. Louis, MO, USA). Antibodies for anti-phospho-ERK-1/2, anti-total ERK-1/2, anti-phospho-p38, anti-total p38, anti-phospho-JNK, anti-total JNK, anti-phospho-IκB, anti-phospho-Akt, anti-Akt, anti-phospho-Src, anti-Src, and anti-Btk were obtained from Cell Signaling Technology Inc. (Beverly, MA, USA). Anti-Phospho-Btk (GTX61791) antibody was obtained from GeneTex (Irvine, CA, USA). Anti-c-Fos, anti-NFATc1, anti-IκB, and anti-PLCγ2 antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-Duoxa1 antibody was obtained from Bioss Inc (BS-11433®, Wobun, MA, USA). Donkey anti-rabbit and anti-mouse immunoglobulin secondary antibodies were purchased from Enzo Life Sciences (Farmingdale, NY, USA).
6
Western Blot Analysis of Cortical Proteins
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The preparation of cortical tissue extraction for immunoblotting was followed by the method described in our previous study63 (link). Protein extracts were subjected to 10% Bis-Tris gel (Invitrogen, Grand Island, NY, USA), incubated with 5% non-fat dry milk in TBST buffer (20 mM Tris-HCl, 150 mM NaCl, 0.1% Tween-20) for 1 h at room temperature, and then followed by the primary antibodies with gently shaking overnight at 4 °C. The primary antibodies used were as follows: anti-EP (Cat# 36-3000, Invitrogen), anti-phos-p38 (Cat# 612288, BD), anti-total-p38 (Cat# 9212, Cell signaling), anti-human Aβ 1-17 clone 6E10 (Cat# 9320-02, Signet), anti-synaptophysin (Cat# MAB5258; Chemicon), anti-synaptojanin 1 antibody (AC1), (Cat# MA3-936; Thermo Fisher), anti-NMDAR2B (Cat# ab81271, Abcam), anti-BACE1 antibody (Cat# ab108394, Abcam), and β-actin (Cat# A5441; Sigma-Aldrich). ImageJ software (National Institutes of Health, Bethesda, MD, USA) was used for quantification of intensity of the immunoreactive bands in the scanned blots.
7
Western Blot Analysis of Signaling Proteins
8
Arctigenin-Induced Cell Apoptosis Signaling
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Arctigenin (Fig. 1 ), 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) and 4´ ,6-diamidino-2-phenylindole dihydrochloride (DAPI) were purchased from Sigma-Aldrich Corp. (St. Louis, MO, USA). LIVE/DEAD Viability/Cytotoxicity kit was purchased from ThermoFisher Scientific, Inc. (Waltham, MA, USA). PhiPhiLux-G1D2 Caspase-3/7 Assay Kit were purchased from OncoImmunin Inc. (Gaithersburg, MD, USA). Anti-cleaved caspase-3, -8, -9, anti-Fas, anti-PARP, anti-Bcl-2, anti-Bcl-xL, anti-Bax, anti-Bad, anti-Phospho-ERK, anti-total-ERK, anti-phospho-p38, anti-total-p38, anti-phospho-JNK, anti-total-JNK, anti-phospho-NFκB, anti-total-NFκB, anti-phospho-AKT, anti-total-AKT, anti-p53 and anti-β-actin antibodies were supplied by Cell Signaling Technology, Inc. (Danvers, MA, USA).
9
Western Blot Analysis of Cellular Signaling
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The cells were lysed on ice using RIPA buffer for 30 minutes by intermittent vortexing and incubation on ice during this period. Protein concentration of whole cell lysates was measured using the Bradford assay (Bio‐Rad). Whole cell lysates (20 μg) for each sample was separated on 8% sodium dodecyl sulfate polyacrylamide gel electrophoresis gel. The separated proteins were transferred onto a polyvinylidene difluoride membrane followed by blocking at room temperature using 5% non‐fat milk diluted in 0.1% TRIS‐buffered saline‐Tween‐20 for an hour. This was followed by an overnight incubation with either anti‐TonEBP (Abcam), anti‐VDR (Santa Cruz), anti‐total p38 (Cell Signaling Technology, Danvers, MA), anti‐phosphorylated p38 (Cell Signaling Technology), anti‐CFTR (Abcam), anti‐tubulin (Cell Signaling Technology), or anti‐beta actin (Santa Cruz) primary antibodies at 4°C. The membranes were washed and incubated with the relevant secondary antibodies conjugated with HRP (anti‐mouse and anti‐rabbit; BosterBio, Pleasanton, CA) for an hour at room temperature. The membranes were washed and incubated with Clarity ECL Western blotting substrate (Bio‐Rad) and resulting chemiluminescence was imaged using Image quant (GE Image Quant LAS 500; GE Healthcare, Chicago, IL). Densitometry analysis was done using ImageJ software (version 6, NIH, Bethesda, MD).
10
Antibody Characterization for TGF-β Signaling
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CG200745 was generously donated by Crystal Genomics Inc. (Seoul, Rep. Korea). Anti-rabbit antibodies against TGF-β1 (polyclonal; Cell Signaling Technology, MA, USA), extracellular signal regulated kinases 1/2 (ERK 1/2), anti-phosphorylated ERK (p-ERK 1/2), anti-c-Jun N-terminal kinase (JNK), anti-phosphorylated JNK (p-JNK), anti-total p38, anti-phosphorylated p38 (p-p38), Smad2/3, and phosphorylated Smad2/3 (Cell Signaling Technology, MA, USA), anti-mouse antibodies against GAPDH (clone GAPDH-71.1;monoclonal), α-SMA (1A4 Clone; monoclonal; Sigma Chemical Co. St. Louis, MO, USA), fibronectin (BD Biosciences, San Jose, CA, USA), heme oxygenase-1 (HO-1, Abcam, Inc., Cambridge, MA, USA), and F4/80 (AbD Serotec, Raleigh, NC, USA) were commercially obtained.
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