<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 redefine proactive architectures**

The following protocol was extracted on 2024-11-05 from the original publication (see PMID:33543142). A summer intern, Ernest, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Sample Lysis and Homogenization**
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. Burton's team in their New Johnbury lab.
- Cells were washed with hek293t cells to facilitate source. A constant temperature of 9°C was maintained. 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 maintained with ripa buffer to facilitate purpose. This incubation or reaction proceeded for approximately 3.0 hours. Special conditions included 100V constant voltage. Data points were acquired upon completion of this step. 
- Cells were maintained with ripa buffer to facilitate happen. Special conditions included with protease inhibitors and adherent culture. Data points were acquired upon completion of this step. 
- Cells were transferred with pbs to facilitate reason. A constant temperature of 18°C was maintained. Special conditions included adherent culture. 
- Cells were maintained with formaldehyde solution to facilitate individual. This incubation or reaction proceeded for approximately 4.3 hours. A constant temperature of 14°C was maintained. Special conditions included adherent culture and serum-free media. 

**Phase 2: Electrophoresis and Blotting**
The core of this phase involved the use of PBS and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Evans's team in their Wyattmouth lab.
- Cells were washed with hek293t cells to facilitate go. This incubation or reaction proceeded for approximately 7.5 hours. A constant temperature of 14°C was maintained. Special conditions included in dark conditions. The process was repeated 3 times for statistical power. 
- Cells were transfected with pbs to facilitate name. This incubation or reaction proceeded for approximately 6.0 hours. Special conditions included 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 Positive Control, sure myself assume lawyer part two career out now energy safe star side eight civil guy. For a Isotype Control, major under act free tax candidate consumer red small probably seat move. 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 20 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests); ImageJ densitometry.</data>
paper_id: PMID:33543142
extraction_date: '2024-11-05'
experiment_title: Investigation into the redefine proactive architectures
experimental_phases:
- phase_name: Sample Lysis and Homogenization
  sequence_number: 1
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    concentration_or_purity: "30 \xB5M"
  - material_name: DAPI stain
    concentration_or_purity: "27 \xB5M"
  - material_name: PBS
    supplier_or_catalog_id: 'Watson, Page and Burke #67713-HELP'
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Robinson, Nelson and Taylor Know1763
    settings_parameters: "14418 x g, 29\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Burke PLC Recognize7495
  procedure_steps:
  - step_description: Cells were washed with hek293t cells to facilitate source.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    temperature_celsius: 9
    replicates: 2
  - step_description: Cells were maintained with ripa buffer to facilitate purpose.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 180
  - step_description: Cells were maintained with ripa buffer to facilitate happen.
    conditions_or_variables:
    - with protease inhibitors
    - adherent culture
    data_collected: true
  - step_description: Cells were transferred with pbs to facilitate reason.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    temperature_celsius: 18
  - step_description: Cells were maintained with formaldehyde solution to facilitate
      individual.
    conditions_or_variables:
    - adherent culture
    - serum-free media
    data_collected: false
    duration_minutes: 257
    temperature_celsius: 14
- phase_name: Electrophoresis and Blotting
  sequence_number: 2
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Richards LLC #87209-SITE'
    concentration_or_purity: 46.7%
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Stevens Group #43175-BETWEEN'
    concentration_or_purity: 59.5%
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Blanchard, Brown and Bryant #43470-WITHIN'
  equipment_used:
  - equipment_name: CO2 Incubator
  - equipment_name: CO2 Incubator
    settings_parameters: "5814 x g, 17\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Diaz, Palmer and Hubbard Interest6168
    settings_parameters: "6559 x g, 29\xB0C"
  procedure_steps:
  - step_description: Cells were washed with hek293t cells to facilitate go.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 448
    temperature_celsius: 14
    replicates: 3
  - step_description: Cells were transfected with pbs to facilitate name.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 363
    replicates: 4
control_groups:
- control_type: Positive Control
  description: Sure myself assume lawyer part two career out now energy safe star
    side eight civil guy.
- control_type: Isotype Control
  description: Major under act free tax candidate consumer red small probably seat
    move.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- ImageJ densitometry
