GC cell lines SGC-7901 was obtained from the cell repository of Chinese Academy of Sciences (Shanghai, China) and cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum, 100 unit/mL penicillin, and 100 μg/mL streptomycin at 37°C in a 5% CO2 incubator. Lapatinib-resistant SGC-7901 cells were developed by 24 h exposure of SGC-7901 cells to 1 μM concentrations of lapatinib. The primary antibodies against HER2, p-HER2, AKT, p-AKT, ERK, and p-ERK were bought from Cell Signaling Technology (Danvers, MA, United States). DMSO, thiazolyl blue tetrazolium bromide (MTT), the anti-β-actin, anti-mouse, and anti-rabbit antibodies were purchased from Sigma (St. Louis, MO, USA). The antibodies against HER-2, p-HER-2, AKT, p-AKT, ERK, p-ERK, and β-actin were from Cell Signaling Technology (Danvers, MA, USA) and Santa Cruz Biotechnology (Santa Cruz, CA, USA).
P erk
Manufactured by Cell Signaling Technology
Sourced in United States, United Kingdom, China, Japan, Canada, Germany, Denmark, Macao
P-ERK is a phospho-specific antibody that recognizes the phosphorylated form of Extracellular Signal-Regulated Kinase (ERK). ERK is a key component of the MAPK signaling pathway, which plays a crucial role in cellular processes such as proliferation, differentiation, and survival. The P-ERK antibody can be used to detect the activated, phosphorylated state of ERK in various experimental applications.
Automatically generated - may contain errors
Lab products found in correlation
P akt, by Cell Signaling Technology (466 mentions)
P jnk, by Cell Signaling Technology (401 mentions)
Gapdh, by Cell Signaling Technology (215 mentions)
β actin, by Cell Signaling Technology (210 mentions)
Cleaved caspase 3, by Cell Signaling Technology (114 mentions)
Bcl 2, by Cell Signaling Technology (110 mentions)
β actin, by Santa Cruz Biotechnology (104 mentions)
Pvdf membrane, by Merck Group (104 mentions)
β actin, by Merck Group (102 mentions)
Caspase 3, by Cell Signaling Technology (89 mentions)
P stat3, by Cell Signaling Technology (84 mentions)
P mek, by Cell Signaling Technology (81 mentions)
Bcl2 associated x (bax), by Cell Signaling Technology (81 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (77 mentions)
P iκbα, by Cell Signaling Technology (66 mentions)
P mtor, by Cell Signaling Technology (57 mentions)
Stat3, by Cell Signaling Technology (57 mentions)
Gapdh, by Santa Cruz Biotechnology (57 mentions)
T erk, by Cell Signaling Technology (48 mentions)
Ire1α, by Cell Signaling Technology (48 mentions)
1 676 protocols using p erk
1
Lapatinib-resistant GC Cell Lines
2
Immunohistochemical Analysis of TNBC
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104 patients diagnosed and treated at the ICO Cancer Center were collected between 1998–2007 among triple negative invasive breast carcinoma (ER/PR and Her2 negative). Representative formalin fixed tumours blocks were selected to establish tissue microarray for 88 patients. For 16 patients, whole tumour block was used as tissue microarray was defective.
Immunolabelling technique was performed by the Benchmark XT automatized tissue staining system (Ventana Medical system) on 4 µm thick blocks section. Primary antibodies used were BCL-XL (BD pharmingen, rabbit polyclonal 556361, CC1 short PH8.4, dilution 1/500), and p-ERK (Phospho-p44/42 MAPK (thr 202/tyr 204) Cell Signaling, rabbit monoclonal, CC1 standard PH8.4, dilution 1/400)
For each staining, the H-Score was calculated as “intensity of staining” x “% of stained cells” where the “intensity of staining” was graded from 0 to 5 (0 none, 1 very weak, 2 weak, 3 intermediate, 4 strong and 5 very strong) and the “percentage of stained cells” estimated from number of tumours cells with cytoplasmic and/or nuclear staining (in case of p-ERK staining) or cytoplasmic staining (in case of BCL-XL staining).
Immunolabelling technique was performed by the Benchmark XT automatized tissue staining system (Ventana Medical system) on 4 µm thick blocks section. Primary antibodies used were BCL-XL (BD pharmingen, rabbit polyclonal 556361, CC1 short PH8.4, dilution 1/500), and p-ERK (Phospho-p44/42 MAPK (thr 202/tyr 204) Cell Signaling, rabbit monoclonal, CC1 standard PH8.4, dilution 1/400)
For each staining, the H-Score was calculated as “intensity of staining” x “% of stained cells” where the “intensity of staining” was graded from 0 to 5 (0 none, 1 very weak, 2 weak, 3 intermediate, 4 strong and 5 very strong) and the “percentage of stained cells” estimated from number of tumours cells with cytoplasmic and/or nuclear staining (in case of p-ERK staining) or cytoplasmic staining (in case of BCL-XL staining).
3
Investigating Lung Cancer Cell Lines
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The human lung cancer cell lines A549, H157, HCC827, and PC9 were obtained from the American Type Culture Collection and were genotyped and authenticated before experiments. Cells were cultured in RPMI-1640 medium (HyClone) supplemented with 10% fetal bovine serum (ZETA) at 37°C in a humidified incubator with 5% CO2. Purified MA (>98%) (#sc-200733) was purchased from Santa Cruz. The 20 mM stock solution was made in DMSO. N-acetyl-l-cysteine (NAC) (#HY-B0215), necrostatin-1 (#HY-15760), Z-VAD-FMK (#HY-16658B), liproxstatin-1 (#HY-12726), erastin (#HY-15763) and osimertinib (#HY-15772) were purchased from MCE. The antibodies used were as follows: p-ERK (#4370), ERK (#4695), p-AMPK (#2535), AMPK (#5832), GPX4 (#52455), SLC7A11 (#12691), NRF2 (#12721), NCOA4 (#66849), FTH1 (#4393), PERK (#5683), IRE1a (#3294), LC3A/B (#12741) and GRP78 (#3177) were purchased from Cell Signaling Technology. BCL2 (#12789-1-AP), KRAS (#12063-1-AP) and GAPDH (#10494-1-AP) were purchased from Proteintech. PLA2G4A (#sc-454) was purchased from Santa Cruz.
4
Comprehensive Signaling Pathway Analysis in NOTCH1-ROS1 Expressing Cells
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NOTCH1–ROS1 expressing NIH3T3 and HEK293 cells were lysed in RIPA buffer and normalized using a Pierce 660-nm Protein Assay. Lysates were run on NuPAGE precast gels (4%–12% Bis-Tris or 3%–8% Tris-Acetate; Invitrogen). For MAPK, PI3K, and JAK/STAT pathway analysis, pMEK (#9154), tMEK (#4694), pERK (#4370), pERK (#9101), tERK (#4695), tERK (#4696), pAKT Thr308 (#4056), pAKT Ser473 (#9271), pAKT Ser473 (#4060), tAKT (#2920), pS6 (#4858), tS6(#2317), pSTAT3 (#9145), tSTAT3 (#9139), pROS1 Tyr2274 (#3078), tROS1 (#15027), and GAPDH (#3683) antibodies were purchased from Cell Signaling (1:1000).
5
Optimized Protocols for Metabolic Studies
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TRIzolTM (catalog #15596018) was purchased from Life Technologies (Carlsbad, CA, USA). A High-Capacity cDNA reverse transcription kit (catalog #4368813) was purchased from Thermo Fisher Scientific (Waltham, MA, USA). A qPCRTM core kit for SYBR Green I (catalog #10-SN10-05) was purchased from Eurogentec (San Diego, CA, USA). 2-NBD glucose (catalog #7133) was purchased from Setareh Biotech (Eugene, OR, USA). Tunicamycin (catalog #T7765), oleate (catalog #O1008), palmitate (catalog #P0500) and sulfo-N-succinimidyl oleate sodium (catalog #SML2148) were purchased from Sigma Aldrich (St. Louis, MO, USA). PERK (catalog #5683), IRE1α (catalog #3294), ATF6 (catalog #65880), BIP (catalog #3177), CHOP (catalog #5554), pAS160 (catalog #8730), pAKT (catalog #4060), PERK (catalog #45899) and ACTIN (catalog #4970) Rabbit mABs were purchased Cell Signaling (Danvers, MA, USA). HRP anti-rabbit secondary IgG antibodies (catalog #ab288151) were purchased from Abcam (Cambridge, MA, USA). All other chemicals and solvents were obtained from Fisher Scientific through its local distributor in the Kingdom of Saudi Arabia.
6
Molecular Mechanisms of Apoptosis Regulation
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Nobiletin, Z-VAD-FMK, Z-DEVD-FMK, dimethyl sulfoxide (DMSO), 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) protease inhibitor cocktail, salubrinal, rabbit anti-human β-actin antibodies, and Dulbecco’s modified Eagle′s medium (DMEM) were purchased from Sigma (St. Louis, MO, USA). An annexin V-FITC/PI apoptosis detection kit was purchased from Pharmingen (San Diego, CA, USA). Antibodies against pro-caspase 9, cleaved caspase 9, caspase 8, pro-caspase3, cleaved caspase 3, Mcl-1, Bax, Bcl-xl, Bad, p-Bad, PARP-1, Bcl-2, 14-3-3, PDI, GRP78, calreticulin, cytochrome C, p-PERK, PERK, p-eIF2α, eIF2 α, ATF6-f, ATF4, IRE-1α, CHOP, p-JNK, p-c-jun, JNK, p-JNK, MAPKp38, p-MAPKp38, ERK, p-ERK, PI3K, p-PI3K, AKT, p-AKT, mTOR, p-mTOR, and goat anti-rabbit and horseradish peroxidase-conjugated immunoglobulin (Ig) G were obtained from Cell Signaling Technology (Danvers, MA, USA). Fetal bovine serum (FBS) was obtained from Biowest (Nuaillé, France).
7
Comprehensive Lung Tumor Analysis
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Lung lobes were fixed in 4% formalin and paraffin embedded. Hematoxylin and eosin staining was performed using standard methods. IHC was performed on 4-μm sections with IHC was performed using Avidin/Biotin Blocking Kit (Vector Laboratories, SP-2001), Avidin-Biotin Complex kit (Vector Laboratories, PK-4001), and DAB Peroxidase Substrate Kit (Vector Laboratories, SK-4100) following standard protocols.
The following primary antibodies were used: Ki-67 (BD Pharmingen, 550609), BrdU (BD Pharmingen, 555627), human mitochondria (Abcam, ab92824), pERK (Cell Signaling, 4370L).
Total tumor burden (tumor area/total area × 100%), mitochondria pos tumor burden (mitochondria pos area/total area × 100%), BrdU pos cell number, Ki67 pos cell number, and pERK pos cell number were calculated using ImageJ.
The following primary antibodies were used: Ki-67 (BD Pharmingen, 550609), BrdU (BD Pharmingen, 555627), human mitochondria (Abcam, ab92824), pERK (Cell Signaling, 4370L).
Total tumor burden (tumor area/total area × 100%), mitochondria pos tumor burden (mitochondria pos area/total area × 100%), BrdU pos cell number, Ki67 pos cell number, and pERK pos cell number were calculated using ImageJ.
8
Comprehensive Lung Tumor Analysis
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Lung lobes were fixed in 4% formalin and paraffin embedded. Hematoxylin and eosin staining was performed using standard methods. IHC was performed on 4-μm sections with IHC was performed using Avidin/Biotin Blocking Kit (Vector Laboratories, SP-2001), Avidin-Biotin Complex kit (Vector Laboratories, PK-4001), and DAB Peroxidase Substrate Kit (Vector Laboratories, SK-4100) following standard protocols.
The following primary antibodies were used: Ki-67 (BD Pharmingen, 550609), BrdU (BD Pharmingen, 555627), human mitochondria (Abcam, ab92824), pERK (Cell Signaling, 4370L).
Total tumor burden (tumor area/total area × 100%), mitochondria pos tumor burden (mitochondria pos area/total area × 100%), BrdU pos cell number, Ki67 pos cell number, and pERK pos cell number were calculated using ImageJ.
The following primary antibodies were used: Ki-67 (BD Pharmingen, 550609), BrdU (BD Pharmingen, 555627), human mitochondria (Abcam, ab92824), pERK (Cell Signaling, 4370L).
Total tumor burden (tumor area/total area × 100%), mitochondria pos tumor burden (mitochondria pos area/total area × 100%), BrdU pos cell number, Ki67 pos cell number, and pERK pos cell number were calculated using ImageJ.
9
Protein Extraction and Western Blot Analysis
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Total protein was extracted from BMMs and C3H10 cells and from the bones of mice used in the OVX experiment using a radioimmunoprecipitation assay lysis buffer (RIPA) (Sigma-Aldrich), and the protein concentration was measured using a bicinchoninic acid protein (BCA) assay kit (EMD Millipore, Bedford, MA, USA). Protein equivalents were separated using 10% SDS PAGE and transferred onto PVDF membranes (EMD Millipore). The membranes were blocked for an hour at room temperature with 5% skim milk in Tris-buffered saline with Tween 20. After incubating primary antibodies overnight at 4 °C, the membranes were nurtured with corresponding HRP conjugated secondary antibodies (Proteintech Group, Chicago, IL, USA.) The bands were visualized employing ECL Luminous Liquid (EMD Millipore). ImageJ software was used to compute the relative grey level of proteins (NIH). Anti-TRAIL was procured from Santa Cruz Bio (Santa Cruz, CA, USA). Anti-CTSK, TRAP, NFATc1, OPG, RUNX2, OCN, RANKL, COL1a, p-P65, P65, p-P38, P38, p-JNK, JNK, p-ERK, ERK, p-IKBα, IKBα, IKKα, IKKβ, and p-IKKα/β were all purchased from Cell Signaling Technology (Danvers, MA, USA).
10
Osteoarthritis Chondrocyte Protein Analysis
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Total lysates of human OA chondrocytes were prepared using RIPA buffer (Biosesang, Gyeonggi-do, Korea). Primary antibodies against MMP3, MMP13 (1:1000 dilution, Cell Signaling Technology, Danvers, MA, USA), COL2A1 (1:1000 dilution, Abcam), ACAN (1:500 dilution, Invitrogen), p38, p-p38, ERK, p-ERK, JNK, p-JNK, p65, p-p65, IκBα, p-IκBα, or β-actin (1:1000 dilution, Cell Signaling Technology) and appropriate HRP-conjugated secondary antibodies (1:2000 dilution, Bethyl) were used. Enhanced chemiluminescence (ECL) detection kit (Thermo Fisher Scientific) was used to detect for specific bands. Chemiluminescent signals were analyzed on a ChemiDoc XRS gel imaging system (Bio-Rad Laboratories, Hercules, CA, USA), and intensity of specific bands was quantified using Image J software (NIH, MD, USA).
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