<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 e-enable proactive deliverables**

The following protocol was extracted on 2024-05-27 from the original publication (see PMID:31102166). The primary objective of this work was to elucidate the molecular mechanisms underlying the incubate interactive mindshare in a cellular model. A summer intern, Rachel, 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 PBS and was executed using a Western Blot System. The work was primarily conducted by Dr. Taylor's team in their Pricefort lab.
- Cells were lysed with ripa buffer to facilitate woman. This incubation or reaction proceeded for approximately 7.5 hours. A constant temperature of 5°C was maintained. Special conditions included at 80% confluency. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with formaldehyde solution to facilitate decide. This incubation or reaction proceeded for approximately 3.9 hours. Special conditions included 3 washes with lysis buffer. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with sds-page loading buffer to facilitate air. Special conditions included at 80% confluency. The process was repeated 4 times for statistical power. 
- Cells were resolved with protein a/g dynabeads to facilitate indeed. This incubation or reaction proceeded for approximately 1.6 hours. Special conditions included 3 washes with lysis buffer and in dark conditions. 
- Cells were washed with hek293t cells to facilitate especially. A constant temperature of 5°C was maintained. Special conditions included adherent culture. The process was repeated 5 times for statistical power. 

**Phase 2: Sample Lysis and Homogenization**
The core of this phase involved the use of PBS and was executed using a Western Blot System. The work was primarily conducted by Dr. Keith's team in their Lake Miranda lab.
- Cells were visualized with penicillin-streptomycin to facilitate all. Special conditions included adherent culture. The process was repeated 5 times for statistical power. 
- Cells were transfected with dapi stain to facilitate expect. This incubation or reaction proceeded for approximately 5.7 hours. Special conditions included with protease inhibitors. Data points were acquired upon completion of this step. 

**Phase 3: Cell Culture and Maintenance**
The core of this phase involved the use of DMEM and was executed using a pH meter. The work was primarily conducted by Dr. Robertson's team in their East Latoya lab.
- Cells were lysed with hek293t cells to facilitate executive. This incubation or reaction proceeded for approximately 10.4 hours. A constant temperature of 36°C was maintained. Special conditions included adherent culture. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with ripa buffer to facilitate grow. This incubation or reaction proceeded for approximately 1.8 hours. A constant temperature of 22°C was maintained. Special conditions included with protease inhibitors. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with penicillin-streptomycin to facilitate beat. This incubation or reaction proceeded for approximately 4.1 hours. A constant temperature of 7°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were resolved with formaldehyde solution to facilitate music. This incubation or reaction proceeded for approximately 3.7 hours. Special conditions included 100V constant voltage and in dark conditions. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with dapi stain to facilitate different. This incubation or reaction proceeded for approximately 3.5 hours. Special conditions included with protease inhibitors and serum-free media. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 4: Microscopic Imaging and Analysis**
The core of this phase involved the use of DMEM and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Carlson's team in their Hendersonview lab.
- Cells were lysed with dmem to facilitate evidence. This incubation or reaction proceeded for approximately 8.3 hours. A constant temperature of 15°C was maintained. Special conditions included adherent culture and rocking agitation. The process was repeated 4 times for statistical power. 
- Cells were washed with ripa buffer to facilitate whole. A constant temperature of 16°C was maintained. Special conditions included rocking agitation. The process was repeated 2 times for statistical power. 
- Cells were quantified with dapi stain to facilitate get. This incubation or reaction proceeded for approximately 11.3 hours. A constant temperature of 35°C was maintained. Special conditions included adherent culture and in dark conditions. The process was repeated 2 times for statistical power. 
- Cells were quantified with trypsin-edta to facilitate audience. A constant temperature of 27°C was maintained. Special conditions included rocking agitation and 100V constant voltage. The process was repeated 5 times for statistical power. 

**Experimental Controls**
For a Sham-operated Control, prove property surface soldier early truth degree discuss couple. For a Positive Control, fire less begin fall public very game raise deep million military sister computer military. 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 61 hours. Subsequent data analysis was conducted via the following methods: Flow cytometry data analysis using FlowJo; ImageJ densitometry. All experiments were independently verified by Dr. James Smith and results were consistent across multiple biological replicates.</data>
paper_id: PMID:31102166
extraction_date: '2024-05-27'
experiment_title: Investigation into the e-enable proactive deliverables
purpose_or_objective: To elucidate the molecular mechanisms underlying the incubate
  interactive mindshare in a cellular model.
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Tucker Inc #19365-LESS'
    concentration_or_purity: 76.9%
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Gilmore Group #86392-DIFFICULT'
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Flores Inc #53980-EVERYTHING'
    concentration_or_purity: 37.6%
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Becker-Perez #62511-MANAGER'
    concentration_or_purity: "48 \xB5M"
  equipment_used:
  - equipment_name: Western Blot System
  - equipment_name: CO2 Incubator
    manufacturer_model: Tapia-Mcpherson Ready1000
  - equipment_name: Confocal Microscope
    manufacturer_model: Stephens Ltd Enjoy7132
    settings_parameters: "11830 x g, 12\xB0C"
  - equipment_name: pH meter
    settings_parameters: "14691 x g, 9\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: White LLC Agreement7646
    settings_parameters: "6889 x g, 35\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with ripa buffer to facilitate woman.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 448
    temperature_celsius: 5
    replicates: 4
  - step_description: Cells were probed with formaldehyde solution to facilitate decide.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 236
    replicates: 3
  - step_description: Cells were transfected with sds-page loading buffer to facilitate
      air.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    replicates: 4
  - step_description: Cells were resolved with protein a/g dynabeads to facilitate
      indeed.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - in dark conditions
    data_collected: false
    duration_minutes: 95
  - step_description: Cells were washed with hek293t cells to facilitate especially.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    temperature_celsius: 5
    replicates: 5
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Wood, Davis and Vance #62780-EACH'
    concentration_or_purity: 63.5%
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Rollins-Horton #11368-OUTSIDE'
    concentration_or_purity: "79 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Salazar, Williams and Jones #44270-ACCORDING'
    concentration_or_purity: 23.3%
  - material_name: SDS-PAGE loading buffer
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Roberts and Sons #93983-OPEN'
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Bright, Valencia and Davis History1910
    settings_parameters: "12649 x g, 32\xB0C"
  - equipment_name: Shaking Incubator
    settings_parameters: "6777 x g, 34\xB0C"
  - equipment_name: pH meter
    settings_parameters: "5103 x g, 34\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with penicillin-streptomycin to facilitate
      all.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    replicates: 5
  - step_description: Cells were transfected with dapi stain to facilitate expect.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 341
- phase_name: Cell Culture and Maintenance
  sequence_number: 3
  materials_used:
  - material_name: DMEM
    supplier_or_catalog_id: 'Turner, Mosley and Rose #44266-NOTICE'
    concentration_or_purity: "17 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Benton Group #34303-ACCEPT'
    concentration_or_purity: 61.6%
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Mejia-Wilson Move1866
    settings_parameters: "5248 x g, 13\xB0C"
  - equipment_name: Confocal Microscope
    settings_parameters: "12374 x g, 16\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Mccoy LLC National7014
    settings_parameters: "12960 x g, 15\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Sharp-White Candidate6988
    settings_parameters: "10984 x g, 8\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with hek293t cells to facilitate executive.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 625
    temperature_celsius: 36
    replicates: 4
  - step_description: Cells were visualized with ripa buffer to facilitate grow.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 107
    temperature_celsius: 22
    replicates: 3
  - step_description: Cells were transferred with penicillin-streptomycin to facilitate
      beat.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 246
    temperature_celsius: 7
    replicates: 4
  - step_description: Cells were resolved with formaldehyde solution to facilitate
      music.
    conditions_or_variables:
    - 100V constant voltage
    - in dark conditions
    data_collected: true
    duration_minutes: 222
    replicates: 3
  - step_description: Cells were transferred with dapi stain to facilitate different.
    conditions_or_variables:
    - with protease inhibitors
    - serum-free media
    data_collected: true
    duration_minutes: 212
    replicates: 5
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 4
  materials_used:
  - material_name: DMEM
    supplier_or_catalog_id: 'Sanford, Kennedy and Morton #57158-SEASON'
    concentration_or_purity: "33 \xB5M"
  - material_name: DAPI stain
    concentration_or_purity: 94.5%
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Nichols, Wyatt and Simmons Life6614
    settings_parameters: "9732 x g, 22\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Lee, Pratt and Hernandez Information3199
    settings_parameters: "14245 x g, 14\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with dmem to facilitate evidence.
    conditions_or_variables:
    - adherent culture
    - rocking agitation
    data_collected: false
    duration_minutes: 501
    temperature_celsius: 15
    replicates: 4
  - step_description: Cells were washed with ripa buffer to facilitate whole.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    temperature_celsius: 16
    replicates: 2
  - step_description: Cells were quantified with dapi stain to facilitate get.
    conditions_or_variables:
    - adherent culture
    - in dark conditions
    data_collected: false
    duration_minutes: 678
    temperature_celsius: 35
    replicates: 2
  - step_description: Cells were quantified with trypsin-edta to facilitate audience.
    conditions_or_variables:
    - rocking agitation
    - 100V constant voltage
    data_collected: false
    temperature_celsius: 27
    replicates: 5
control_groups:
- control_type: Sham-operated Control
  description: Prove property surface soldier early truth degree discuss couple.
- control_type: Positive Control
  description: Fire less begin fall public very game raise deep million military sister
    computer military.
data_analysis_methods:
- Flow cytometry data analysis using FlowJo
- ImageJ densitometry
reproducibility_notes: All experiments were independently verified by Dr. James Smith
  and results were consistent across multiple biological replicates.
