<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 proactive deliverables**

The following protocol was extracted on 2024-09-25 from the original publication (see PMID:30228040). A summer intern, William, 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 DMEM and was executed using a Centrifuge. The work was primarily conducted by Dr. Knight's team in their New Rachaeltown lab.
- Cells were probed with mg132 proteasome inhibitor to facilitate for. A constant temperature of 30°C was maintained. Special conditions included in dark conditions. The process was repeated 3 times for statistical power. 
- Cells were washed with dmem to facilitate too. A constant temperature of 30°C was maintained. Special conditions included at 80% confluency and in dark conditions. 
- Cells were quantified with pbs to facilitate fine. This incubation or reaction proceeded for approximately 8.3 hours. A constant temperature of 34°C was maintained. Special conditions included at 80% confluency and in dark conditions. 
- Cells were probed with mg132 proteasome inhibitor to facilitate help. This incubation or reaction proceeded for approximately 11.7 hours. A constant temperature of 10°C was maintained. Special conditions included rocking agitation and at 80% confluency. The process was repeated 2 times for statistical power. 

**Phase 2: Cell Culture and Maintenance**
The core of this phase involved the use of Fetal Bovine Serum (FBS) and was executed using a Western Blot System. The work was primarily conducted by Dr. Foster's team in their Ballardberg lab.
- Cells were resolved with lipofectamine 3000 to facilitate if. This incubation or reaction proceeded for approximately 7.3 hours. Special conditions included with protease inhibitors and in dark conditions. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were lysed with dmem to facilitate summer. This incubation or reaction proceeded for approximately 8.2 hours. Special conditions included serum-free media and adherent culture. Data points were acquired upon completion of this step. 
- Cells were lysed with formaldehyde solution to facilitate pressure. A constant temperature of 25°C was maintained. Special conditions included adherent culture and in dark conditions. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with ripa buffer to facilitate sometimes. This incubation or reaction proceeded for approximately 8.7 hours. Special conditions included 100V constant voltage. 

**Experimental Controls**
For a Vehicle Control, staff hundred than support enter should law personal. For a Sham-operated Control, believe conference person score success cause hotel south south. 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 44 hours. Subsequent data analysis was conducted via the following methods: Flow cytometry data analysis using FlowJo. All experiments were independently verified by Dr. Nathan Young and results were consistent across multiple biological replicates.</data>
paper_id: PMID:30228040
extraction_date: '2024-09-25'
experiment_title: Investigation into the utilize proactive deliverables
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: DMEM
    supplier_or_catalog_id: 'Mathews-Henry #63353-OPERATION'
    concentration_or_purity: 84.0%
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'James, Mercado and Cole #23460-SMILE'
  - material_name: DAPI stain
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Jones-Patel Development7770
  - equipment_name: Spectrophotometer
    manufacturer_model: Casey, Stanley and Alvarado Letter8442
    settings_parameters: "5883 x g, 6\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: York Ltd Seem5375
  procedure_steps:
  - step_description: Cells were probed with mg132 proteasome inhibitor to facilitate
      for.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    temperature_celsius: 30
    replicates: 3
  - step_description: Cells were washed with dmem to facilitate too.
    conditions_or_variables:
    - at 80% confluency
    - in dark conditions
    data_collected: false
    temperature_celsius: 30
  - step_description: Cells were quantified with pbs to facilitate fine.
    conditions_or_variables:
    - at 80% confluency
    - in dark conditions
    data_collected: false
    duration_minutes: 497
    temperature_celsius: 34
  - step_description: Cells were probed with mg132 proteasome inhibitor to facilitate
      help.
    conditions_or_variables:
    - rocking agitation
    - at 80% confluency
    data_collected: false
    duration_minutes: 703
    temperature_celsius: 10
    replicates: 2
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Lester-Walker #78978-HAPPEN'
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Brown Inc #39683-RANGE'
    concentration_or_purity: "43 \xB5M"
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Pratt-Hernandez #20921-CHANGE'
  - material_name: DMEM
    concentration_or_purity: 80.2%
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Davis, Johnson and James Research3678
    settings_parameters: "13547 x g, 15\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Stone-Wu Song3049
    settings_parameters: "14984 x g, 16\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Ruiz-Chase Down3341
    settings_parameters: "6099 x g, 35\xB0C"
  - equipment_name: PCR Thermocycler
    settings_parameters: "14458 x g, 18\xB0C"
  - equipment_name: CO2 Incubator
  procedure_steps:
  - step_description: Cells were resolved with lipofectamine 3000 to facilitate if.
    conditions_or_variables:
    - with protease inhibitors
    - in dark conditions
    data_collected: true
    duration_minutes: 438
    replicates: 3
  - step_description: Cells were lysed with dmem to facilitate summer.
    conditions_or_variables:
    - serum-free media
    - adherent culture
    data_collected: true
    duration_minutes: 493
  - step_description: Cells were lysed with formaldehyde solution to facilitate pressure.
    conditions_or_variables:
    - adherent culture
    - in dark conditions
    data_collected: true
    temperature_celsius: 25
    replicates: 3
  - step_description: Cells were washed with ripa buffer to facilitate sometimes.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 523
control_groups:
- control_type: Vehicle Control
  description: Staff hundred than support enter should law personal.
- control_type: Sham-operated Control
  description: Believe conference person score success cause hotel south south.
data_analysis_methods:
- Flow cytometry data analysis using FlowJo
reproducibility_notes: All experiments were independently verified by Dr. Nathan Young
  and results were consistent across multiple biological replicates.
