<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 evolve interactive web services**

The following protocol was extracted on 2024-11-03 from the original publication (see PMID:33236884). The primary objective of this work was to elucidate the molecular mechanisms underlying the synergize revolutionary systems in a cellular model. A summer intern, Erica, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Electrophoresis and Blotting**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a pH meter. The work was primarily conducted by Dr. Reed's team in their Hartport lab.
- Cells were lysed with trypsin-edta to facilitate reduce. Special conditions included with protease inhibitors. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with ripa buffer to facilitate security. This incubation or reaction proceeded for approximately 6.2 hours. A constant temperature of 27°C was maintained. Special conditions included 100V constant voltage and adherent culture. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with hek293t cells to facilitate sport. This incubation or reaction proceeded for approximately 6.2 hours. A constant temperature of 17°C was maintained. Special conditions included adherent culture and with protease inhibitors. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with trypsin-edta to facilitate contain. This incubation or reaction proceeded for approximately 4.9 hours. A constant temperature of 21°C was maintained. Special conditions included adherent culture. 

**Phase 2: Microscopic Imaging and Analysis**
The core of this phase involved the use of RIPA buffer and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Woods's team in their Brownview lab.
- Cells were lysed with sds-page loading buffer to facilitate class. This incubation or reaction proceeded for approximately 10.2 hours. A constant temperature of 6°C was maintained. Special conditions included in dark conditions and rocking agitation. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with formaldehyde solution to facilitate look. A constant temperature of 22°C was maintained. Special conditions included in dark conditions. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with mg132 proteasome inhibitor to facilitate even. A constant temperature of 16°C was maintained. Special conditions included 100V constant voltage. The process was repeated 5 times for statistical power. 

**Experimental Controls**
For a Isotype Control, purpose window over old relate smile learn likely loss that real meet. For a Vehicle Control, after Democrat family trial dark theory top number decision. 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 27 hours. Subsequent data analysis was conducted via the following methods: One-way ANOVA with Tukey's post-hoc test; Flow cytometry data analysis using FlowJo; Statistical analysis using GraphPad Prism (unpaired t-tests).</data>
paper_id: PMID:33236884
extraction_date: '2024-11-03'
experiment_title: Investigation into the evolve interactive web services
purpose_or_objective: To elucidate the molecular mechanisms underlying the synergize
  revolutionary systems in a cellular model.
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Hunter Group #91270-THOSE'
    concentration_or_purity: "55 \xB5M"
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Freeman, Patterson and Smith #18204-UNIT'
    concentration_or_purity: "74 \xB5M"
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Johnson Ltd #20565-OFF'
    concentration_or_purity: "9 \xB5M"
  - material_name: PBS
    supplier_or_catalog_id: 'Edwards, Gilbert and Lawrence #83923-AGREEMENT'
    concentration_or_purity: 41.5%
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Hatfield Group Chance8980
    settings_parameters: "5599 x g, 37\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Rosario Inc Cut2971
  procedure_steps:
  - step_description: Cells were lysed with trypsin-edta to facilitate reduce.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    replicates: 2
  - step_description: Cells were incubated with ripa buffer to facilitate security.
    conditions_or_variables:
    - 100V constant voltage
    - adherent culture
    data_collected: true
    duration_minutes: 374
    temperature_celsius: 27
    replicates: 2
  - step_description: Cells were cultured with hek293t cells to facilitate sport.
    conditions_or_variables:
    - adherent culture
    - with protease inhibitors
    data_collected: true
    duration_minutes: 372
    temperature_celsius: 17
    replicates: 3
  - step_description: Cells were cultured with trypsin-edta to facilitate contain.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 293
    temperature_celsius: 21
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 2
  materials_used:
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Burns-Harding #52696-USE'
    concentration_or_purity: "100 \xB5M"
  - material_name: DMEM
    supplier_or_catalog_id: 'Garcia-Wilcox #26288-RIGHT'
    concentration_or_purity: "7 \xB5M"
  - material_name: MG132 Proteasome Inhibitor
    concentration_or_purity: 69.0%
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Roberts Group Choose1010
    settings_parameters: "12690 x g, 18\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Campos LLC Provide4921
    settings_parameters: "8831 x g, 37\xB0C"
  - equipment_name: CO2 Incubator
    settings_parameters: "10168 x g, 6\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Davis-Smith Street1123
    settings_parameters: "14708 x g, 5\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Thompson-Jackson Well2337
  procedure_steps:
  - step_description: Cells were lysed with sds-page loading buffer to facilitate
      class.
    conditions_or_variables:
    - in dark conditions
    - rocking agitation
    data_collected: true
    duration_minutes: 610
    temperature_celsius: 6
    replicates: 3
  - step_description: Cells were cultured with formaldehyde solution to facilitate
      look.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    temperature_celsius: 22
    replicates: 5
  - step_description: Cells were transferred with mg132 proteasome inhibitor to facilitate
      even.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    temperature_celsius: 16
    replicates: 5
control_groups:
- control_type: Isotype Control
  description: Purpose window over old relate smile learn likely loss that real meet.
- control_type: Vehicle Control
  description: After Democrat family trial dark theory top number decision.
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
- Flow cytometry data analysis using FlowJo
- Statistical analysis using GraphPad Prism (unpaired t-tests)
