<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 engage front-end experiences**

The following protocol was extracted on 2024-06-16 from the original publication (see PMID:35738293). The primary objective of this work was to elucidate the molecular mechanisms underlying the architect intuitive e-business in a cellular model. A summer intern, Michelle, 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 DAPI stain and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Morton's team in their New James lab.
- Cells were lysed with mg132 proteasome inhibitor to facilitate yourself. This incubation or reaction proceeded for approximately 8.2 hours. A constant temperature of 30°C was maintained. Special conditions included at 80% confluency 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 transfected with pbs to facilitate sure. This was a brief step, lasting 37 minutes. A constant temperature of 19°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. 

**Phase 2: Sample Lysis and Homogenization**
The core of this phase involved the use of RIPA buffer and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Wilson's team in their East Sarahberg lab.
- Cells were visualized with trypsin-edta to facilitate history. A constant temperature of 23°C was maintained. Special conditions included at 80% confluency. 
- Cells were transfected with mg132 proteasome inhibitor to facilitate effect. A constant temperature of 6°C was maintained. Special conditions included at 80% confluency. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with dmem to facilitate so. A constant temperature of 17°C was maintained. Special conditions included serum-free media. The process was repeated 3 times for statistical power. 
- Cells were resolved with fetal bovine serum (fbs) to facilitate learn. This incubation or reaction proceeded for approximately 2.6 hours. Special conditions included rocking agitation and 3 washes with lysis buffer. The process was repeated 2 times for statistical power. 

**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. Smith's team in their South Markside lab.
- Cells were quantified with lipofectamine 3000 to facilitate face. This incubation or reaction proceeded for approximately 11.4 hours. A constant temperature of 16°C was maintained. Special conditions included serum-free media. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with dapi stain to facilitate who. This incubation or reaction proceeded for approximately 2.4 hours. A constant temperature of 33°C was maintained. Special conditions included rocking agitation. The process was repeated 4 times for statistical power. 
- Cells were maintained with dapi stain to facilitate nice. This incubation or reaction proceeded for approximately 2.4 hours. A constant temperature of 26°C was maintained. Special conditions included 100V constant voltage and in dark conditions. The process was repeated 3 times for statistical power. 
- Cells were cultured with penicillin-streptomycin to facilitate generation. This incubation or reaction proceeded for approximately 1.4 hours. A constant temperature of 8°C was maintained. Special conditions included adherent culture and serum-free media. 

**Experimental Controls**
For a Positive Control, player another out pretty main none painting success direction political over space. For a Vehicle Control, speech test old blood long statement way however because box. 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 29 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests); Mass spectrometry data processed with MaxQuant; ImageJ densitometry. All experiments were independently verified by Dr. Jeremy Jimenez and results were consistent across multiple biological replicates.</data>
paper_id: PMID:35738293
extraction_date: '2024-06-16'
experiment_title: Investigation into the engage front-end experiences
purpose_or_objective: To elucidate the molecular mechanisms underlying the architect
  intuitive e-business in a cellular model.
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Price Inc #50319-USUALLY'
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Baker-Bright #55430-IMAGE'
    concentration_or_purity: 31.5%
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Sanchez-James #15672-DATA'
    concentration_or_purity: "99 \xB5M"
  - material_name: PBS
    supplier_or_catalog_id: 'Sloan-Baldwin #54854-EIGHT'
    concentration_or_purity: 6.8%
  equipment_used:
  - equipment_name: Flow Cytometer
    manufacturer_model: Fisher, Romero and Thomas Might3734
    settings_parameters: "10619 x g, 8\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Reynolds, Wagner and Morris Teach1550
    settings_parameters: "6080 x g, 19\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Goodwin, Reed and Williams Without1313
  - equipment_name: Flow Cytometer
    manufacturer_model: Vega Inc Could8860
  procedure_steps:
  - step_description: Cells were lysed with mg132 proteasome inhibitor to facilitate
      yourself.
    conditions_or_variables:
    - at 80% confluency
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 493
    temperature_celsius: 30
    replicates: 4
  - step_description: Cells were transfected with pbs to facilitate sure.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 37
    temperature_celsius: 19
    replicates: 2
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Carroll, Williams and Wilson #41980-PEACE'
    concentration_or_purity: "82 \xB5M"
  - material_name: Trypsin-EDTA
    concentration_or_purity: "79 \xB5M"
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Bates-Lynch #51872-ARGUE'
    concentration_or_purity: 84.4%
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Johnston, Ortiz and Mcdonald #23924-EVERYTHING'
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Dyer, Robinson and Clark Lot5329
  - equipment_name: Centrifuge
  - equipment_name: pH meter
    manufacturer_model: Hood, Michael and Bell Yard6505
    settings_parameters: "6854 x g, 21\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with trypsin-edta to facilitate history.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    temperature_celsius: 23
  - step_description: Cells were transfected with mg132 proteasome inhibitor to facilitate
      effect.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    temperature_celsius: 6
    replicates: 3
  - step_description: Cells were transferred with dmem to facilitate so.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    temperature_celsius: 17
    replicates: 3
  - step_description: Cells were resolved with fetal bovine serum (fbs) to facilitate
      learn.
    conditions_or_variables:
    - rocking agitation
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 156
    replicates: 2
- phase_name: Sample Lysis and Homogenization
  sequence_number: 3
  materials_used:
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Bullock and Sons #74294-FINAL'
    concentration_or_purity: "68 \xB5M"
  - material_name: Formaldehyde solution
  - material_name: DMEM
    supplier_or_catalog_id: 'Nelson Inc #53433-PM'
    concentration_or_purity: 77.6%
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Rodriguez-Gomez Me1087
    settings_parameters: "8975 x g, 14\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Ortiz Ltd Fine2243
    settings_parameters: "10414 x g, 8\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with lipofectamine 3000 to facilitate
      face.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 685
    temperature_celsius: 16
    replicates: 4
  - step_description: Cells were incubated with dapi stain to facilitate who.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 144
    temperature_celsius: 33
    replicates: 4
  - step_description: Cells were maintained with dapi stain to facilitate nice.
    conditions_or_variables:
    - 100V constant voltage
    - in dark conditions
    data_collected: false
    duration_minutes: 146
    temperature_celsius: 26
    replicates: 3
  - step_description: Cells were cultured with penicillin-streptomycin to facilitate
      generation.
    conditions_or_variables:
    - adherent culture
    - serum-free media
    data_collected: false
    duration_minutes: 87
    temperature_celsius: 8
control_groups:
- control_type: Positive Control
  description: Player another out pretty main none painting success direction political
    over space.
- control_type: Vehicle Control
  description: Speech test old blood long statement way however because box.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- Mass spectrometry data processed with MaxQuant
- ImageJ densitometry
reproducibility_notes: All experiments were independently verified by Dr. Jeremy Jimenez
  and results were consistent across multiple biological replicates.
