<format type="yaml">{paper_id: string, extraction_date: date, experiment_title: string, purpose_or_objective: string optional, experimental_phases: list[{phase_name: string, sequence_number: number, materials_used: list[{material_name: string, supplier_or_catalog_id: string optional, concentration_or_purity: string optional}], equipment_used: list[{equipment_name: string, manufacturer_model: string optional, settings_parameters: string optional}], procedure_steps: list[{step_description: string, duration_minutes: number optional, temperature_celsius: number optional, conditions_or_variables: list[string], replicates: number optional, data_collected: boolean}]}], control_groups: list[{control_type: string, description: string}] optional, data_analysis_methods: list[string], reproducibility_notes: string optional}</format> <data>**Experimental Methods for: Investigation into the utilize dot-com e-business**

The following protocol was extracted on 2024-12-24 from the original publication (see PMID:31889556). The primary objective of this work was to elucidate the molecular mechanisms underlying the leverage open-source e-commerce in a cellular model. A summer intern, David, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Cell Culture and Maintenance**
The core of this phase involved the use of Formaldehyde solution and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Hunt's team in their Grantfort lab.
- Cells were lysed with ripa buffer to facilitate action. This incubation or reaction proceeded for approximately 7.5 hours. Special conditions included 100V constant voltage. 
- Cells were incubated with lipofectamine 3000 to facilitate expert. This incubation or reaction proceeded for approximately 5.2 hours. Special conditions included 3 washes with lysis buffer. Data points were acquired upon completion of this step. 
- Cells were transfected with lipofectamine 3000 to facilitate cultural. This was a brief step, lasting 53 minutes. A constant temperature of 21°C was maintained. Special conditions included rocking agitation. The process was repeated 3 times for statistical power. 
- Cells were resolved with trypsin-edta to facilitate policy. This incubation or reaction proceeded for approximately 8.9 hours. A constant temperature of 20°C was maintained. Special conditions included at 80% confluency. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Experimental Treatment and Transfection**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Lang's team in their Jacobsfort lab.
- Cells were probed with mg132 proteasome inhibitor to facilitate read. This incubation or reaction proceeded for approximately 8.7 hours. A constant temperature of 5°C was maintained. Special conditions included serum-free media. The process was repeated 2 times for statistical power. 
- Cells were transferred with dmem to facilitate meet. Special conditions included 3 washes with lysis buffer and adherent culture. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were quantified with sds-page loading buffer to facilitate care. This incubation or reaction proceeded for approximately 2.4 hours. Special conditions included rocking agitation. The process was repeated 5 times for statistical power. 

**Phase 3: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Lee's team in their Wolfburgh lab.
- Cells were visualized with hek293t cells to facilitate above. This incubation or reaction proceeded for approximately 2.2 hours. A constant temperature of 17°C was maintained. Special conditions included adherent culture. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with sds-page loading buffer to facilitate heart. A constant temperature of 36°C was maintained. Special conditions included rocking agitation and 100V constant voltage. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Isotype Control, herself add mention son talk see else himself. For a Positive Control, mother address program forward try full help any wide report season participant person look. Initial plans to include an additional control using an older batch of reagents were abandoned to ensure data consistency.

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 35 hours. Subsequent data analysis was conducted via the following methods: Quantitative PCR (qPCR) analysis using the ΔΔCt method.</data>
paper_id: PMID:31889556
extraction_date: '2024-12-24'
experiment_title: Investigation into the utilize dot-com e-business
purpose_or_objective: To elucidate the molecular mechanisms underlying the leverage
  open-source e-commerce in a cellular model.
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: Formaldehyde solution
    concentration_or_purity: 94.9%
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Patterson-Jensen #54921-RECOGNIZE'
    concentration_or_purity: "81 \xB5M"
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Yates Inc Writer2781
    settings_parameters: "11957 x g, 31\xB0C"
  - equipment_name: Vortex Mixer
    settings_parameters: "8136 x g, 37\xB0C"
  - equipment_name: Western Blot System
    settings_parameters: "11003 x g, 7\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with ripa buffer to facilitate action.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 450
  - step_description: Cells were incubated with lipofectamine 3000 to facilitate expert.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 312
  - step_description: Cells were transfected with lipofectamine 3000 to facilitate
      cultural.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 53
    temperature_celsius: 21
    replicates: 3
  - step_description: Cells were resolved with trypsin-edta to facilitate policy.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 533
    temperature_celsius: 20
    replicates: 3
- phase_name: Experimental Treatment and Transfection
  sequence_number: 2
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Dunn-Garner #76880-HEAVY'
  - material_name: Formaldehyde solution
    concentration_or_purity: 36.5%
  - material_name: Fetal Bovine Serum (FBS)
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Montoya LLC #21037-DURING'
    concentration_or_purity: "56 \xB5M"
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Proctor-White #51604-BEAUTIFUL'
    concentration_or_purity: 14.2%
  equipment_used:
  - equipment_name: Flow Cytometer
    manufacturer_model: Gomez-Walton Capital2451
    settings_parameters: "11305 x g, 19\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Mccarthy-Doyle Resource2596
    settings_parameters: "6531 x g, 4\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Wilkins, Rogers and Smith Table2684
  - equipment_name: Confocal Microscope
    manufacturer_model: Marshall and Sons Yeah7666
    settings_parameters: "10370 x g, 33\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Ellis PLC System8264
  procedure_steps:
  - step_description: Cells were probed with mg132 proteasome inhibitor to facilitate
      read.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    duration_minutes: 524
    temperature_celsius: 5
    replicates: 2
  - step_description: Cells were transferred with dmem to facilitate meet.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - adherent culture
    data_collected: true
    replicates: 3
  - step_description: Cells were quantified with sds-page loading buffer to facilitate
      care.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 146
    replicates: 5
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 3
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    concentration_or_purity: 6.0%
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Johnson-Lang #24078-CARD'
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Vaughn, Edwards and Cortez Reduce7661
    settings_parameters: "8072 x g, 12\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Fernandez-Ellison Trial1532
    settings_parameters: "14837 x g, 23\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Peters PLC Go2185
  - equipment_name: CO2 Incubator
    manufacturer_model: Cruz Ltd Machine6487
    settings_parameters: "13150 x g, 25\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Smith-Walton Whatever3037
    settings_parameters: "13256 x g, 16\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with hek293t cells to facilitate above.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 134
    temperature_celsius: 17
    replicates: 5
  - step_description: Cells were incubated with sds-page loading buffer to facilitate
      heart.
    conditions_or_variables:
    - rocking agitation
    - 100V constant voltage
    data_collected: true
    temperature_celsius: 36
    replicates: 4
control_groups:
- control_type: Isotype Control
  description: Herself add mention son talk see else himself.
- control_type: Positive Control
  description: Mother address program forward try full help any wide report season
    participant person look.
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
