Dulbecco’s Modified Eagle Medium (DMEM) with high glucose was from Invitrogen. The HCC827, H292, H4006, 293T cell lines were purchased from American Type Culture Collection. Enzastaurin and erlotinib were from LC Laboratories (Woburn, MA). Go6976 and U0126 were purchased from EMD Millipore (Billerica, MA). Anti-phospho-p70S6K (Thr389), S6K, mTOR, phospho- Erk½ (Thr202/Tyr204), Erk½, (E747-A750del Specific) EGFR, EGFR, P-Akt (Ser473), P-Akt (Thr308), Akt, PKCα and ErbB3 antibodies were from Cell Signaling Technology (Danvers, MA), and anti-actin antibody was purchased from Sigma-Aldrich (St. Louis, MO). Anti-EEA1 antibody was from BD Biosciences (San Jose, CA). siRNAs for PKCα and Gab1 were from Qiagen (Valencia, CA) and those for PLCγ1 were from ambion (life technologies). Dual mTOR inhibitor (KU-0063794) and Akt inhibitor (MK-2206 2HCl) were purchased from Selleckchem. All other reagents were from Sigma-Aldrich (St. Louis, MO).
Anti eea1 antibody
Manufactured by BD
The Anti-EEA1 antibody is a laboratory reagent used for the detection and identification of the early endosomal antigen 1 (EEA1) protein in various cell and tissue samples. It is a specific and sensitive tool for researchers studying endosomal trafficking, cell signaling, and related cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Anti actin antibody, by Merck Group (3 mentions)
U0126, by Merck Group (3 mentions)
Erbb3, by Cell Signaling Technology (2 mentions)
Dulbecco s modified eagle medium dmem with high glucose, by Thermo Fisher Scientific (2 mentions)
Go6976, by Merck Group (2 mentions)
Sirnas for pkcα and gab1, by Qiagen (2 mentions)
Those for plcγ1, by Thermo Fisher Scientific (2 mentions)
Dual mtor inhibitor ku 0063794, by Selleck Chemicals (2 mentions)
Akt inhibitor mk 2206 2hcl, by Selleck Chemicals (2 mentions)
Erlotinib, by LC Laboratories (2 mentions)
Enzastaurin, by LC Laboratories (2 mentions)
Hcc827, by American Type Culture Collection (2 mentions)
H4006, by American Type Culture Collection (2 mentions)
Ab7280, by Abcam (1 mentions)
Ab83232, by Abcam (1 mentions)
Ab40776, by Abcam (1 mentions)
Ab7771, by Abcam (1 mentions)
Anti lamp1 antibody, by Cell Signaling Technology (1 mentions)
Anti erk antibody, by Cell Signaling Technology (1 mentions)
Anti p erk antibody, by Cell Signaling Technology (1 mentions)
6 protocols using anti eea1 antibody
1
Investigating EGFR Signaling Pathways
2
Western Blot and Immunofluorescence Protocols
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WB experiments were performed as previously reported [69 (link),93 (link)] using the following antibodies: anti-β-Actin antibody (4967; Cell Signaling Technology, Danvers, Massachusetts), anti-NICD antibody (ab83232; Abcam, Cambridge, UK), anti-Myc antibody (06–549; Millipore), anti-Flag antibody (F7425; Sigma-Aldrich), anti-DLL4 antibody (ab7280; Abcam), anti-JAG1 antibody (ab7771; Abcam), anti-Notch1 antibody (3608; Cell Signaling Technology), anti-Erk antibody (9102; Cell Signaling Technology), anti-p-Erk antibody (9101; Cell Signaling Technology), anti-p-AKT antibody (4060; Cell Signaling Technology), and anti-AKT antibody (9272; Cell Signaling Technology). Quantification of protein level using gray analysis (Gel-Pro analyzer). Immunofluorescence experiments were performed as previously reported [94 (link)] using the following antibodies: anti-Notch1 antibody (3608; Cell Signaling Technology), anti-Notch1 antibody (3447; Cell Signaling Technology), anti-DLL4 antibody (ab7280; Abcam), anti-JAG1 antibody (ab7771; Abcam), anti-LAMP1 antibody (15665; Cell Signaling Technology), anti-EEA1 antibody (610457; BD Biosciences, Franklin Lakes, New Jersey), anti-APPL1 antibody (3858; Cell Signaling Technology), anti-AKT antibody (2920; Cell Signaling Technology), anti-AKT antibody (9272; Cell Signaling Technology), and anti-PIK3CA antibody (ab40776; Abcam).
3
Investigating EGFR Signaling Pathways
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Dulbecco’s Modified Eagle Medium (DMEM) with high glucose was from Invitrogen. The HCC827, H292, H4006, 293T cell lines were purchased from American Type Culture Collection. Enzastaurin and erlotinib were from LC Laboratories (Woburn, MA). Go6976 and U0126 were purchased from EMD Millipore (Billerica, MA). Anti-phospho-p70S6K (Thr389), S6K, mTOR, phospho- Erk½ (Thr202/Tyr204), Erk½, (E747-A750del Specific) EGFR, EGFR, P-Akt (Ser473), P-Akt (Thr308), Akt, PKCα and ErbB3 antibodies were from Cell Signaling Technology (Danvers, MA), and anti-actin antibody was purchased from Sigma-Aldrich (St. Louis, MO). Anti-EEA1 antibody was from BD Biosciences (San Jose, CA). siRNAs for PKCα and Gab1 were from Qiagen (Valencia, CA) and those for PLCγ1 were from ambion (life technologies). Dual mTOR inhibitor (KU-0063794) and Akt inhibitor (MK-2206 2HCl) were purchased from Selleckchem. All other reagents were from Sigma-Aldrich (St. Louis, MO).
4
Receptor Agonist Synthesis and Antibody Procurement
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PAR1 peptide agonist (SFLLRN) and PAR2 peptide agonist (SLIGKV) were synthesized and purified by reverse-phase, high-pressure liquid chromatography at the Tufts University Core Facility (Boston, MA). Epidermal growth factor was purchased from Sigma-Aldrich (St. Louis, MO). Polyclonal anti-FLAG and anti-HA antibodies were purchased from Rockland Immunochemicals (Pottstown, PA). Anti-EEA1 antibody was obtained from BD Biosciences (East Rutherford, NJ). Monoclonal anti-PAR1 WEDE antibody was purchased from Beckman Coulter (Brea, CA). Polyclonal anti-ARRDC3, monoclonal anti-ALIX, and monoclonal anti-Ub (P4D1) antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Anti-EGFR antibody was obtained from Cell Signaling Technologies (Boston, MA). Monoclonal anti-HA antibody was purchased from Covance (San Diego, CA). Horseradish peroxidase–conjugated goat anti-rabbit and goat anti-mouse antibodies were purchased from Bio-Rad Laboratories (Hercules, CA). Alexa Fluor 488, 594, and 647 secondary antibodies were purchased from Invitrogen (Carlsbad, CA).
5
Regulation of mTOR Signaling by PKC
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All cell lines were purchased from American Tissue Culture Collection (Manassas, VA, USA). Lipofectamine RNAiMAX were from Life Technologies (Grand Island, NY). X-treme GENE 9 were from Roche (Indianapolis, IN). PLD2 inhibitor, VU 0364739 was a kind gift from Dr. Alex Brown (Vanderbilt University, Nashville, TN, USA). PMA, Gö 6976, Bisindolylmaleimide I and U0126 were from EMD Millipore (Billerica, MA). Anti-phospho-p70S6K (Thr389), S6K, mTOR, phospho-ERK1/2 (Thr202/Tyr204), ERK1/2, EGFR, HA antibodies were from Cell Signaling Technology (Danvers, MA) and anti-Actin antibody was purchased from Sigma Aldrich (St. Louis, MO). Anti-LAMP1, LAMP2, PKCδ and PKCε antibodies were from Santa Cruz Biotechnology (Santa Cruz, CA). Anti-EEA1 antibody was from BD Biosciences (San Jose, CA). siRNA for PKCη, PKCδ, PKCε, RAGB, LAMTOR 1, and LAMTOR 3 were from Qiagen (Valencia, CA).
6
Visualizing CAR-T Cell Tumor Infiltration
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For the tumor infiltrating CAR-T cells detection, a fraction of tumor tissues were fixed in 4% paraformaldehyde, then dehydrated in 30% sucrose and embedded in OCT compound (SAKURA #4583) followed by cryosection generation on a cryostat (Leica). Images were collected using 20 3 objective of the TCS SP8 STED microscope (Leica). The imaging of lysosome and mitochondrial in CAR-T cells was performed in living cells using LysoTracker Red (Invitrogen #L7528) and MitoTracker Orange (Invitrogen #M7510). For imaging analysis of CAR-EGFP and lysosomes, CAR-T cells were first co-incubated with CD19 + K562 cells for 30 min. For imaging analysis of CAR-EGFP and mCherry-TRAF2 within the early endosomes, CAR-T cells were fixed using 4% paraformaldehyde and permeabilized with 0.1% Triton X-100 in PBS containing 1% BSA. Early endosome was labeled using anti-EEA1 antibody (BD Bioscience #610456). Images of cell samples were acquired by A1R-si (Nikon) or TCS SP8 STED microscope with 60 3 and 63 3 oil immersion objective. Captured images were analyzed with ImageJ software (NIH).
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