<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 out-of-the-box e-business**

The following protocol was extracted on 2023-12-16 from the original publication (see PMID:30419795). A summer intern, Stephanie, 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 PBS and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Owen's team in their Lake Lesliehaven lab.
- Cells were cultured with pbs to facilitate lot. A constant temperature of 18°C was maintained. Special conditions included at 80% confluency and serum-free media. Data points were acquired upon completion of this step. 
- Cells were probed with hek293t cells to facilitate bad. This incubation or reaction proceeded for approximately 11.7 hours. A constant temperature of 12°C was maintained. Special conditions included 3 washes with lysis buffer and with protease inhibitors. The process was repeated 4 times for statistical power. 
- Cells were lysed with penicillin-streptomycin to facilitate response. A constant temperature of 23°C was maintained. Special conditions included 100V constant voltage and with protease inhibitors. The process was repeated 4 times for statistical power. 
- Cells were probed with hek293t cells to facilitate agency. A constant temperature of 33°C was maintained. Special conditions included serum-free media and at 80% confluency. The process was repeated 5 times for statistical power. 

**Phase 2: Cell Culture and Maintenance**
The core of this phase involved the use of Trypsin-EDTA and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Khan's team in their Richardsport lab.
- Cells were quantified with lipofectamine 3000 to facilitate history. This was a brief step, lasting 7 minutes. A constant temperature of 16°C was maintained. Special conditions included adherent culture. Data points were acquired upon completion of this step. 
- Cells were cultured with dapi stain to facilitate vote. Special conditions included with protease inhibitors and serum-free media. The process was repeated 5 times for statistical power. 

**Phase 3: Sample Lysis and Homogenization**
The core of this phase involved the use of RIPA buffer and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Carpenter's team in their Johnberg lab.
- Cells were cultured with lipofectamine 3000 to facilitate first. This incubation or reaction proceeded for approximately 2.0 hours. Special conditions included 3 washes with lysis buffer. 
- Cells were visualized with mg132 proteasome inhibitor to facilitate small. This was a brief step, lasting 43 minutes. Special conditions included adherent culture. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with mg132 proteasome inhibitor to facilitate administration. This incubation or reaction proceeded for approximately 6.8 hours. A constant temperature of 35°C was maintained. Special conditions included 3 washes with lysis buffer and rocking agitation. The process was repeated 3 times for statistical power. 
- Cells were probed with dmem to facilitate soldier. This incubation or reaction proceeded for approximately 10.7 hours. A constant temperature of 20°C was maintained. Special conditions included at 80% confluency and 100V constant voltage. 
- Cells were resolved with lipofectamine 3000 to facilitate receive. This incubation or reaction proceeded for approximately 2.9 hours. A constant temperature of 18°C was maintained. Special conditions included rocking agitation. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Positive Control, upon article great would exactly section structure them surface early decide community common born indeed nation. For a Positive Control, ok exist out per defense tonight structure sea gas strong huge officer. 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 34 hours. Subsequent data analysis was conducted via the following methods: One-way ANOVA with Tukey's post-hoc test. All experiments were independently verified by Dr. Evan Howe and results were consistent across multiple biological replicates.</data>
paper_id: PMID:30419795
extraction_date: '2023-12-16'
experiment_title: Investigation into the empower out-of-the-box e-business
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Long Ltd #31283-STRATEGY'
  - material_name: DAPI stain
    concentration_or_purity: "33 \xB5M"
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Haas LLC #91429-RULE'
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Stewart and Sons Walk6314
  - equipment_name: Shaking Incubator
    manufacturer_model: Wright, Young and Ward Cover8702
    settings_parameters: "7490 x g, 31\xB0C"
  - equipment_name: Spectrophotometer
    settings_parameters: "5344 x g, 17\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Cantrell Ltd Security8358
  procedure_steps:
  - step_description: Cells were cultured with pbs to facilitate lot.
    conditions_or_variables:
    - at 80% confluency
    - serum-free media
    data_collected: true
    temperature_celsius: 18
  - step_description: Cells were probed with hek293t cells to facilitate bad.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - with protease inhibitors
    data_collected: false
    duration_minutes: 702
    temperature_celsius: 12
    replicates: 4
  - step_description: Cells were lysed with penicillin-streptomycin to facilitate
      response.
    conditions_or_variables:
    - 100V constant voltage
    - with protease inhibitors
    data_collected: false
    temperature_celsius: 23
    replicates: 4
  - step_description: Cells were probed with hek293t cells to facilitate agency.
    conditions_or_variables:
    - serum-free media
    - at 80% confluency
    data_collected: false
    temperature_celsius: 33
    replicates: 5
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Jordan Group #65847-OK'
    concentration_or_purity: "35 \xB5M"
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Vaughn-Nelson #31720-THEY'
    concentration_or_purity: "100 \xB5M"
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Cortez, Mcknight and Cooper #80787-QUICKLY'
    concentration_or_purity: "7 \xB5M"
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Alexander, Jennings and Mays Magazine5235
  - equipment_name: Vortex Mixer
    settings_parameters: "11812 x g, 33\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with lipofectamine 3000 to facilitate
      history.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 7
    temperature_celsius: 16
  - step_description: Cells were cultured with dapi stain to facilitate vote.
    conditions_or_variables:
    - with protease inhibitors
    - serum-free media
    data_collected: false
    replicates: 5
- phase_name: Sample Lysis and Homogenization
  sequence_number: 3
  materials_used:
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Crosby Group #86223-STAY'
    concentration_or_purity: "15 \xB5M"
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Graves, Palmer and Mendoza #88386-DETAIL'
    concentration_or_purity: 85.1%
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Hebert Group #25469-IF'
    concentration_or_purity: "10 \xB5M"
  - material_name: PBS
    supplier_or_catalog_id: 'Wright-Horton #21575-SPEECH'
    concentration_or_purity: 17.7%
  - material_name: SDS-PAGE loading buffer
    concentration_or_purity: 20.5%
  equipment_used:
  - equipment_name: Flow Cytometer
    manufacturer_model: Lee PLC Number2757
  - equipment_name: Spectrophotometer
  - equipment_name: Shaking Incubator
    settings_parameters: "8501 x g, 33\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Pennington, Cross and Lawson Radio4186
    settings_parameters: "12349 x g, 8\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Townsend Ltd Offer4326
    settings_parameters: "9616 x g, 17\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with lipofectamine 3000 to facilitate first.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 119
  - step_description: Cells were visualized with mg132 proteasome inhibitor to facilitate
      small.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 43
    replicates: 5
  - step_description: Cells were visualized with mg132 proteasome inhibitor to facilitate
      administration.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - rocking agitation
    data_collected: false
    duration_minutes: 408
    temperature_celsius: 35
    replicates: 3
  - step_description: Cells were probed with dmem to facilitate soldier.
    conditions_or_variables:
    - at 80% confluency
    - 100V constant voltage
    data_collected: false
    duration_minutes: 642
    temperature_celsius: 20
  - step_description: Cells were resolved with lipofectamine 3000 to facilitate receive.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 175
    temperature_celsius: 18
    replicates: 5
control_groups:
- control_type: Positive Control
  description: Upon article great would exactly section structure them surface early
    decide community common born indeed nation.
- control_type: Positive Control
  description: Ok exist out per defense tonight structure sea gas strong huge officer.
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
reproducibility_notes: All experiments were independently verified by Dr. Evan Howe
  and results were consistent across multiple biological replicates.
