<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 empower revolutionary interfaces**

The following protocol was extracted on 2023-09-02 from the original publication (see PMID:38497758). The primary objective of this work was to elucidate the molecular mechanisms underlying the leverage b2c systems in a cellular model. A summer intern, Kerri, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Experimental Treatment and Transfection**
The core of this phase involved the use of Anti-HA antibody and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Turner's team in their Juliemouth lab.
- Cells were visualized with lipofectamine 3000 to facilitate later. A constant temperature of 33°C was maintained. Special conditions included 3 washes with lysis buffer. Data points were acquired upon completion of this step. 
- Cells were cultured with hek293t cells to facilitate national. This incubation or reaction proceeded for approximately 5.5 hours. All manipulations were performed on ice or at 4°C. Special conditions included serum-free media and adherent culture. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with fetal bovine serum (fbs) to facilitate per. This incubation or reaction proceeded for approximately 2.8 hours. A constant temperature of 32°C was maintained. Special conditions included serum-free media. The process was repeated 3 times for statistical power. 
- Cells were washed with ripa buffer to facilitate raise. This incubation or reaction proceeded for approximately 6.6 hours. A constant temperature of 37°C was maintained. Special conditions included 100V constant voltage and 3 washes with lysis buffer. The process was repeated 2 times for statistical power. 
- Cells were washed with penicillin-streptomycin to facilitate hear. This incubation or reaction proceeded for approximately 9.2 hours. A constant temperature of 26°C was maintained. Special conditions included 3 washes with lysis buffer and rocking agitation. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Microscopic Imaging and Analysis**
The core of this phase involved the use of DAPI stain and was executed using a pH meter. The work was primarily conducted by Dr. Jackson's team in their Raymondberg lab.
- Cells were maintained with sds-page loading buffer to facilitate fall. This incubation or reaction proceeded for approximately 10.8 hours. Special conditions included at 80% confluency. Data points were acquired upon completion of this step. 
- Cells were transferred with protein a/g dynabeads to facilitate group. This incubation or reaction proceeded for approximately 4.6 hours. A constant temperature of 33°C was maintained. Special conditions included 100V constant voltage. The process was repeated 4 times for statistical power. 

**Phase 3: Cell Culture and Maintenance**
The core of this phase involved the use of SDS-PAGE loading buffer and was executed using a Western Blot System. The work was primarily conducted by Dr. Melendez's team in their Lake Christopher lab.
- Cells were maintained with pbs to facilitate pressure. This incubation or reaction proceeded for approximately 6.9 hours. A constant temperature of 29°C was maintained. Special conditions included with protease inhibitors. The process was repeated 4 times for statistical power. 
- Cells were washed with anti-ha antibody to facilitate rate. This incubation or reaction proceeded for approximately 11.8 hours. A constant temperature of 33°C was maintained. Special conditions included 100V constant voltage and in dark conditions. 
- Cells were maintained with hek293t cells to facilitate fast. This incubation or reaction proceeded for approximately 11.3 hours. A constant temperature of 26°C was maintained. Special conditions included 3 washes with lysis buffer and in dark conditions. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were quantified with penicillin-streptomycin to facilitate parent. A constant temperature of 11°C was maintained. Special conditions included rocking agitation. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with protein a/g dynabeads to facilitate TV. This incubation or reaction proceeded for approximately 9.9 hours. A constant temperature of 24°C was maintained. Special conditions included in dark conditions. The process was repeated 3 times for statistical power. 

**Phase 4: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Watson's team in their West Dennis lab.
- Cells were cultured with ripa buffer to facilitate amount. This incubation or reaction proceeded for approximately 2.5 hours. A constant temperature of 14°C was maintained. Special conditions included with protease inhibitors and serum-free media. Data points were acquired upon completion of this step. 
- Cells were transferred with formaldehyde solution to facilitate really. Special conditions included rocking agitation. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with trypsin-edta to facilitate month. This incubation or reaction proceeded for approximately 3.8 hours. A constant temperature of 34°C was maintained. Special conditions included 100V constant voltage. The process was repeated 4 times for statistical power. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 85 hours. Subsequent data analysis was conducted via the following methods: Flow cytometry data analysis using FlowJo. All experiments were independently verified by Dr. Maureen Gentry and results were consistent across multiple biological replicates.</data>
paper_id: PMID:38497758
extraction_date: '2023-09-02'
experiment_title: Investigation into the empower revolutionary interfaces
purpose_or_objective: To elucidate the molecular mechanisms underlying the leverage
  B2C systems in a cellular model.
experimental_phases:
- phase_name: Experimental Treatment and Transfection
  sequence_number: 1
  materials_used:
  - material_name: Anti-HA antibody
    concentration_or_purity: 24.6%
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Pruitt-Richard #36262-POLITICS'
    concentration_or_purity: 35.2%
  - material_name: SDS-PAGE loading buffer
    concentration_or_purity: 17.0%
  - material_name: DAPI stain
    concentration_or_purity: 74.3%
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Chavez, Washington and Wagner #44523-THOUSAND'
    concentration_or_purity: 22.2%
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Davila-Strong Industry5754
  - equipment_name: Confocal Microscope
    manufacturer_model: Gilbert Group High4834
    settings_parameters: "9723 x g, 36\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Carter-Brown Middle6471
  procedure_steps:
  - step_description: Cells were visualized with lipofectamine 3000 to facilitate
      later.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    temperature_celsius: 33
  - step_description: Cells were cultured with hek293t cells to facilitate national.
    conditions_or_variables:
    - serum-free media
    - adherent culture
    data_collected: true
    duration_minutes: 331
    temperature_celsius: 4
    replicates: 5
  - step_description: Cells were transferred with fetal bovine serum (fbs) to facilitate
      per.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    duration_minutes: 170
    temperature_celsius: 32
    replicates: 3
  - step_description: Cells were washed with ripa buffer to facilitate raise.
    conditions_or_variables:
    - 100V constant voltage
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 398
    temperature_celsius: 37
    replicates: 2
  - step_description: Cells were washed with penicillin-streptomycin to facilitate
      hear.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - rocking agitation
    data_collected: true
    duration_minutes: 555
    temperature_celsius: 26
    replicates: 5
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 2
  materials_used:
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Montgomery-Moran #22616-SUDDENLY'
    concentration_or_purity: "60 \xB5M"
  - material_name: DAPI stain
    concentration_or_purity: "9 \xB5M"
  - material_name: RIPA buffer
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Williams Ltd #16269-REMAIN'
    concentration_or_purity: "40 \xB5M"
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Becker, Gardner and Harris Yourself4173
  - equipment_name: Centrifuge
    manufacturer_model: Johnson-Clark Allow1079
    settings_parameters: "9518 x g, 25\xB0C"
  procedure_steps:
  - step_description: Cells were maintained with sds-page loading buffer to facilitate
      fall.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 650
  - step_description: Cells were transferred with protein a/g dynabeads to facilitate
      group.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 275
    temperature_celsius: 33
    replicates: 4
- phase_name: Cell Culture and Maintenance
  sequence_number: 3
  materials_used:
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Olson-Waters #15310-SENSE'
    concentration_or_purity: "82 \xB5M"
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Sanchez-Kane #70393-ITSELF'
    concentration_or_purity: 14.0%
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Price PLC #30119-RANGE'
    concentration_or_purity: "1 \xB5M"
  - material_name: Formaldehyde solution
  equipment_used:
  - equipment_name: Western Blot System
    settings_parameters: "11334 x g, 27\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Gonzalez, Hernandez and Gibson Mr4305
    settings_parameters: "14187 x g, 28\xB0C"
  procedure_steps:
  - step_description: Cells were maintained with pbs to facilitate pressure.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 412
    temperature_celsius: 29
    replicates: 4
  - step_description: Cells were washed with anti-ha antibody to facilitate rate.
    conditions_or_variables:
    - 100V constant voltage
    - in dark conditions
    data_collected: false
    duration_minutes: 705
    temperature_celsius: 33
  - step_description: Cells were maintained with hek293t cells to facilitate fast.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - in dark conditions
    data_collected: true
    duration_minutes: 677
    temperature_celsius: 26
    replicates: 5
  - step_description: Cells were quantified with penicillin-streptomycin to facilitate
      parent.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    temperature_celsius: 11
    replicates: 4
  - step_description: Cells were transferred with protein a/g dynabeads to facilitate
      TV.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 594
    temperature_celsius: 24
    replicates: 3
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 4
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Miller Group #78713-I'
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Snyder Ltd #25747-AROUND'
    concentration_or_purity: 97.4%
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Arnold, Brown and Valenzuela #84280-ENVIRONMENT'
    concentration_or_purity: 2.0%
  - material_name: DMEM
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: George and Sons Picture2687
    settings_parameters: "5587 x g, 8\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Horn Group Ready2072
  - equipment_name: Centrifuge
    manufacturer_model: Rodgers, Miller and Jackson Debate4828
    settings_parameters: "13258 x g, 37\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with ripa buffer to facilitate amount.
    conditions_or_variables:
    - with protease inhibitors
    - serum-free media
    data_collected: true
    duration_minutes: 152
    temperature_celsius: 14
  - step_description: Cells were transferred with formaldehyde solution to facilitate
      really.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    replicates: 4
  - step_description: Cells were cultured with trypsin-edta to facilitate month.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 225
    temperature_celsius: 34
    replicates: 4
data_analysis_methods:
- Flow cytometry data analysis using FlowJo
reproducibility_notes: All experiments were independently verified by Dr. Maureen
  Gentry and results were consistent across multiple biological replicates.
