GAPDH (6c5) and AR (N-20) antibodies were purchased from Santa Cruz Biotechnology. Phospho-p38 (Thr180/Tyr182) antibody was from Cell Signaling Technology. AR antibody for IP was from Millipore. The p38 and Phospho-Hsp27 (ser78) antibody were purchased from One World Lab. Anti-mouse/rabbit second antibody for Western Blot was from Invitrogen. The p38 inhibitor (SB-203580) was from Apexbio. Doc was from Fisher Scientific.
Phospho p38 thr180 tyr182 antibody
Manufactured by Cell Signaling Technology
Sourced in United States
The Phospho-p38 (Thr180/Tyr182) antibody is a specific detection reagent for the phosphorylated form of the p38 mitogen-activated protein kinase. It recognizes the p38 protein only when phosphorylated at threonine 180 and tyrosine 182, which are required for its activation. This antibody can be used in various immunoassays to detect and quantify the activated p38 MAPK.
Automatically generated - may contain errors
Lab products found in correlation
P38 antibody, by Cell Signaling Technology (3 mentions)
Ar n 20 antibodies, by Santa Cruz Biotechnology (2 mentions)
Ar antibody for ip, by Merck Group (2 mentions)
P38 inhibitor sb 203580, by Apexbio (2 mentions)
Gapdh 6c5, by Santa Cruz Biotechnology (2 mentions)
Anti mouse rabbit second antibody for western blot, by Thermo Fisher Scientific (2 mentions)
Hematoxylin, by Merck Group (2 mentions)
Anti chemerin antibody, by Abcam (2 mentions)
Envison system hrp dab for mouse primary antibodies, by Agilent Technologies (2 mentions)
Vector dab peroxidase hrp substrate kit, by Vector Laboratories (2 mentions)
Signalstain boost ihc detection reagent, by Cell Signaling Technology (2 mentions)
Rarres2 antibody, by Abcam (2 mentions)
Goat anti mouse igg horseradish peroxidase hrp, by Santa Cruz Biotechnology (2 mentions)
Phospho β catenin ser33 37 thr41 antibody, by Cell Signaling Technology (2 mentions)
Gapdh antibody, by Santa Cruz Biotechnology (2 mentions)
β catenin antibody, by Cell Signaling Technology (2 mentions)
Goat anti rabbit igg hrp, by Santa Cruz Biotechnology (2 mentions)
Halt phosphatase inhibitor cocktail, by Thermo Fisher Scientific (2 mentions)
Halt protease inhibitor cocktail, by Thermo Fisher Scientific (2 mentions)
Ly294002, by Cell Signaling Technology (1 mentions)
9 protocols using phospho p38 thr180 tyr182 antibody
1
Western Blot Antibody Validation
2
Immunohistochemistry for Tissue Protein Expression
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Tissue-section slides were de-paraffinized in xylene and rehydrated in ethanol, followed by antigen retrieval with 10% citrate buffer (pH 6.0) in a pressure cooker for 10 minutes at 120 °C. Slides were incubated with the primary antibody at 4°C overnight. For RARRES2 staining, tissues were fixed with 10% formalin and embedded in paraffin. Anti-chemerin antibody (Abcam, Cambridge, MA, ab72965) was used at 12.5 μg/ml. Immunostaining was performed using the Dako Envison+ System-HRP (DAB) for mouse primary antibodies (Dako, Carpinteria, CA). IHC slides were scored using the formula Total staining intensity = Σ (intensity scores × percent positivity), where intensity score 0 = no staining, 1 = weak staining, 2 = moderate staining, 3 = strong staining. Interpreter of the IHC was blind to the diagnosis and gene expression levels accessed by real-time qPCR. For phospho-p38 staining, primary antibody used was phospho-p38 (Thr180/Tyr182) antibody (1: 800, Cell Signaling, Danvers, MA, #4511). Immunostaining was performed by detection with SignalStain Boost IHC detection reagents (HRP, Cell signaling, rabbit #8114), followed by staining with Vector DAB peroxidase (HRP) substrate kit (Vector laboratories, Burlingame, CA, SK-4100). Hematoxylin (Sigma-Aldrich, St. Louis, MO) was used for counterstaining.
3
Antibody Characterization for Cell Signaling Assays
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Phospho‐Akt (Ser473 and Thr308) antibody, phospho‐GSK‐3α (Ser21)/GSK‐3β (Ser9) antibody, phospho‐p38 (Thr180/Tyr182) antibody, Akt antibody, GSK‐3α/β antibody, p38 antibody, and glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) antibody were purchased from Cell Signaling (Danvers, MA, USA). α‐SMA antibody was purchased from Sigma‐Aldrich (St. Louis, MO, USA). Alexa Fluor 488 goat anti‐mouse immunoglobulin G (IgG) and 4′,6‐diamidino‐2‐phenylindole (DAPI) were purchased from Invitrogen (Carlsbad, CA, USA). ML221 was purchased from Sigma‐Aldrich. LY294002 was purchased from Cell Signaling.
4
Immunohistochemistry for Tissue Protein Expression
5
Exosome Protein Expression Analysis
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Samples (exosomes or K562 cells) with equal protein concentration (50 μg) were subjected to SDS-PAGE and transferred to PVDF membrane (PerkinElmer, Life Science). Blots were blocked with 5% skim milk-TBS-Tween 20 for 1 h at room temperature and probed with primary antibodies against target proteins overnight at 4°C. Primary antibodies LMO2 (1:1000) (EP3257, catalogue no # GTX62229), TSG101 (1:1000) (4A10, catalogue no # GTX70255) were purchased from GeneTex. hnRNPA1 antibody (1:1000) (D21H11, catalogue no # 8443), Akt (1:2000) (C67E7, catalogue no #4691), Phospho-Akt (Ser473) (1:2000) (D9E, catalogue no #4060), p38 MAPK antibody (1:1000) (catalogue no # 9212) and Phospho-p38 (Thr180/Tyr182) antibody (1:2000) (28B10, catalogue no # 9216) were purchased from Cell Signaling. CD63 antibody (1:500) (NKI/C-3, catalogue no # MA5-11501) was purchased from ThermoScientific. γ-globin antibody was purchased from Abcam (1:1000) (EPR9708, catalogue no ab137096). β-actin antibody was purchased from Sigma-Aldrich (1:4000) (AC-15, catalogue no A5441). Followed by which blots were washed with PBST and incubated with horseradish peroxidase-conjugated secondary antibodies (1:1000) for 1 h at room temperature. The blots were washed and incubated in ECL solution (Thermo, cat. 32209) for 1 min and then exposed by ImageQuant LAS4000 (GE Healthcare).
6
Protein Expression and Signaling Analysis
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A lysis buffer with 1% of SDS,10 mM of Tris HCl (pH 7.4), and supplemented with 1× Halt protease inhibitor cocktail and 1× Halt phosphatase inhibitor cocktail (Thermo Fisher Scientific) was used. The primary antibodies used were: RARRES2 antibody (1:1 000, Abcam, ab72965); GAPDH antibody (1: 5 000, Santa Cruz Biotechnology, sc-47724), β-catenin antibody (1:1 000, Cell Signaling, #8480), phospho-β-catenin (Ser33/37/Thr41) antibody (1: 1 000, Cell signaling, #9561), phospho-p38 (Thr180/Tyr182) antibody (1:1 000, Cell Signaling, #4511), p38 antibody (1:1 000, Cell Signaling, #8690). Secondary antibodies used were goat anti-mouse IgG-horseradish peroxidase (HRP) and goat anti-rabbit IgG-HRP (1:5 000, Santa Cruz Biotechnology). Except for Western blots including multiple tissue samples, each Western blot experiments were repeated three times with independent sample sets.
7
Protein Expression and Signaling Analysis
8
Western Blot Antibody Validation
9
Notch Signaling Regulation in LPS-Induced
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LPS from Escherichia coli (0111:B4), AA and DAPT were purchased from Sigma–Aldrich. Dll4 was obtained from R&D Systems. β-Actin and Hes1 primary antibody were obtained from Abcam, Notch3 and iNOS primary antibody were from Santa Cruz Biotechnology. Phospho-C-Jun N-terminal kinase (JNK) (Thr183/Tyr185) antibody, phospho-extracellular signal-regulated kinase (ERK)1/2 (Thr202/Tyr204) antibody, phospho-p38 (Thr180/Tyr182) antibody, and phospho-IκBα antibody were purchased from Cell Signaling Technology. All secondary antibodies were from Boster Biological Technology. All other reagents were from commercial suppliers and of standard biochemical quality.
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