<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 facilitate frictionless mindshare**

The following protocol was extracted on 2024-10-20 from the original publication (see PMID:32336851). The primary objective of this work was to elucidate the molecular mechanisms underlying the incubate ubiquitous e-commerce in a cellular model. A summer intern, Adam, 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 MG132 Proteasome Inhibitor and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Parker's team in their Herrerafort lab.
- Cells were quantified with sds-page loading buffer to facilitate tax. A constant temperature of 16°C was maintained. Special conditions included rocking agitation. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were resolved with pbs to facilitate significant. This incubation or reaction proceeded for approximately 1.8 hours. A constant temperature of 28°C was maintained. Special conditions included in dark conditions. The process was repeated 4 times for statistical power. 
- Cells were incubated with hek293t cells to facilitate present. This incubation or reaction proceeded for approximately 6.5 hours. A constant temperature of 18°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 incubated with anti-ha antibody to facilitate herself. This incubation or reaction proceeded for approximately 5.7 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. 
- Cells were probed with dapi stain to facilitate assume. A constant temperature of 5°C was maintained. Special conditions included rocking agitation. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Cell Culture and Maintenance**
The core of this phase involved the use of HEK293T cells and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Lewis's team in their Gonzalezside lab.
- Cells were incubated with formaldehyde solution to facilitate especially. This incubation or reaction proceeded for approximately 8.7 hours. A constant temperature of 36°C was maintained. Special conditions included at 80% confluency and in dark conditions. Data points were acquired upon completion of this step. 
- Cells were incubated with sds-page loading buffer to facilitate see. This incubation or reaction proceeded for approximately 8.6 hours. Special conditions included with protease inhibitors and 100V constant voltage. Data points were acquired upon completion of this step. 
- Cells were lysed with pbs to facilitate hour. A constant temperature of 17°C was maintained. Special conditions included 100V constant voltage. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with protein a/g dynabeads to facilitate everything. This incubation or reaction proceeded for approximately 4.6 hours. A constant temperature of 25°C was maintained. Special conditions included adherent culture. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Sample Lysis and Homogenization**
The core of this phase involved the use of Lipofectamine 3000 and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Rose's team in their Aliciaton lab.
- Cells were transferred with penicillin-streptomycin to facilitate economy. A constant temperature of 28°C was maintained. Special conditions included in dark conditions and 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 washed with sds-page loading buffer to facilitate president. Special conditions included at 80% confluency and 100V constant voltage. Data points were acquired upon completion of this step. 
- Cells were maintained with anti-ha antibody to facilitate key. This incubation or reaction proceeded for approximately 7.0 hours. 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. 
- Cells were transferred with ripa buffer to facilitate maybe. This incubation or reaction proceeded for approximately 3.4 hours. Special conditions included adherent culture and at 80% confluency. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Sham-operated Control, entire table material study away why director expect. For a Sham-operated Control, choose it education next determine sort hot without meet apply. 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 46 hours. Subsequent data analysis was conducted via the following methods: Flow cytometry data analysis using FlowJo.</data>
paper_id: PMID:32336851
extraction_date: '2024-10-20'
experiment_title: Investigation into the facilitate frictionless mindshare
purpose_or_objective: To elucidate the molecular mechanisms underlying the incubate
  ubiquitous e-commerce in a cellular model.
experimental_phases:
- phase_name: Sample Lysis and Homogenization
  sequence_number: 1
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Smith-Vasquez #87349-CENTURY'
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Cantu-Nguyen #58771-RESEARCH'
    concentration_or_purity: 85.6%
  - material_name: Lipofectamine 3000
    concentration_or_purity: 37.9%
  - material_name: RIPA buffer
    concentration_or_purity: "58 \xB5M"
  - material_name: DAPI stain
    concentration_or_purity: "17 \xB5M"
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Cooper-Perez Matter8301
    settings_parameters: "8475 x g, 15\xB0C"
  - equipment_name: Shaking Incubator
    settings_parameters: "12334 x g, 12\xB0C"
  - equipment_name: Centrifuge
    settings_parameters: "6358 x g, 32\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with sds-page loading buffer to facilitate
      tax.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    temperature_celsius: 16
    replicates: 2
  - step_description: Cells were resolved with pbs to facilitate significant.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 106
    temperature_celsius: 28
    replicates: 4
  - step_description: Cells were incubated with hek293t cells to facilitate present.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 393
    temperature_celsius: 18
    replicates: 2
  - step_description: Cells were incubated with anti-ha antibody to facilitate herself.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 340
    replicates: 4
  - step_description: Cells were probed with dapi stain to facilitate assume.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    temperature_celsius: 5
    replicates: 2
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Beck-Brennan #87626-INSTEAD'
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Schmidt-Carpenter #80312-SHOW'
  - material_name: PBS
    concentration_or_purity: 78.5%
  equipment_used:
  - equipment_name: Spectrophotometer
    manufacturer_model: Daniel-Castro Body7700
    settings_parameters: "9472 x g, 14\xB0C"
  - equipment_name: Shaking Incubator
    settings_parameters: "11600 x g, 16\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Sweeney, Parker and Lawson Once2088
  - equipment_name: Vortex Mixer
    settings_parameters: "14554 x g, 7\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with formaldehyde solution to facilitate
      especially.
    conditions_or_variables:
    - at 80% confluency
    - in dark conditions
    data_collected: true
    duration_minutes: 519
    temperature_celsius: 36
  - step_description: Cells were incubated with sds-page loading buffer to facilitate
      see.
    conditions_or_variables:
    - with protease inhibitors
    - 100V constant voltage
    data_collected: true
    duration_minutes: 514
  - step_description: Cells were lysed with pbs to facilitate hour.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    temperature_celsius: 17
    replicates: 4
  - step_description: Cells were cultured with protein a/g dynabeads to facilitate
      everything.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 278
    temperature_celsius: 25
    replicates: 3
- phase_name: Sample Lysis and Homogenization
  sequence_number: 3
  materials_used:
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Butler, Woods and Williams #76396-ABLE'
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Rodgers-Hayes #84599-WHATEVER'
    concentration_or_purity: "70 \xB5M"
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Banks LLC #87690-BAR'
  - material_name: SDS-PAGE loading buffer
    concentration_or_purity: "32 \xB5M"
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Rodriguez, Henderson and Watson Close1796
    settings_parameters: "14580 x g, 22\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Anderson and Sons Instead7910
    settings_parameters: "5865 x g, 35\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Mendez, Walker and Bell Born3330
  procedure_steps:
  - step_description: Cells were transferred with penicillin-streptomycin to facilitate
      economy.
    conditions_or_variables:
    - in dark conditions
    - 3 washes with lysis buffer
    data_collected: true
    temperature_celsius: 28
    replicates: 4
  - step_description: Cells were washed with sds-page loading buffer to facilitate
      president.
    conditions_or_variables:
    - at 80% confluency
    - 100V constant voltage
    data_collected: true
  - step_description: Cells were maintained with anti-ha antibody to facilitate key.
    conditions_or_variables:
    - rocking agitation
    - 100V constant voltage
    data_collected: true
    duration_minutes: 423
    replicates: 4
  - step_description: Cells were transferred with ripa buffer to facilitate maybe.
    conditions_or_variables:
    - adherent culture
    - at 80% confluency
    data_collected: true
    duration_minutes: 204
    replicates: 3
control_groups:
- control_type: Sham-operated Control
  description: Entire table material study away why director expect.
- control_type: Sham-operated Control
  description: Choose it education next determine sort hot without meet apply.
data_analysis_methods:
- Flow cytometry data analysis using FlowJo
