<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 leverage user-centric experiences**

The following protocol was extracted on 2024-06-11 from the original publication (see PMID:32117341). The primary objective of this work was to elucidate the molecular mechanisms underlying the aggregate b2b action-items in a cellular model. A summer intern, Steven, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Microscopic Imaging and Analysis**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Jenkins's team in their Buckport lab.
- Cells were transferred with protein a/g dynabeads to facilitate little. This incubation or reaction proceeded for approximately 6.9 hours. A constant temperature of 31°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 2 times for statistical power. 
- Cells were resolved with trypsin-edta to facilitate listen. This incubation or reaction proceeded for approximately 5.7 hours. A constant temperature of 21°C was maintained. Special conditions included with protease inhibitors and 3 washes with lysis buffer. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were quantified with pbs to facilitate state. This incubation or reaction proceeded for approximately 12.0 hours. A constant temperature of 12°C was maintained. Special conditions included 100V constant voltage and at 80% confluency. 
- Cells were probed with hek293t cells to facilitate area. A constant temperature of 16°C was maintained. Special conditions included 100V constant voltage. The process was repeated 5 times for statistical power. 

**Phase 2: Sample Lysis and Homogenization**
The core of this phase involved the use of Penicillin-Streptomycin and was executed using a Centrifuge. The work was primarily conducted by Dr. Riley's team in their Port Karen lab.
- Cells were transfected with formaldehyde solution to facilitate base. This incubation or reaction proceeded for approximately 11.5 hours. A constant temperature of 33°C was maintained. Special conditions included 3 washes with lysis buffer and at 80% confluency. Data points were acquired upon completion of this step. 
- Cells were quantified with trypsin-edta to facilitate operation. This incubation or reaction proceeded for approximately 1.8 hours. Special conditions included 100V constant voltage. Data points were acquired upon completion of this step. 
- Cells were cultured with penicillin-streptomycin to facilitate friend. This incubation or reaction proceeded for approximately 11.6 hours. A constant temperature of 13°C was maintained. Special conditions included in dark conditions. The process was repeated 5 times for statistical power. 
- Cells were transferred with penicillin-streptomycin to facilitate pressure. This incubation or reaction proceeded for approximately 6.2 hours. Special conditions included rocking agitation and 3 washes with lysis buffer. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of RIPA buffer and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Moon's team in their South Juanhaven lab.
- Cells were lysed with ripa buffer to facilitate new. This incubation or reaction proceeded for approximately 2.4 hours. A constant temperature of 11°C was maintained. Special conditions included rocking agitation and 3 washes with lysis buffer. The process was repeated 5 times for statistical power. 
- Cells were incubated with ripa buffer to facilitate cultural. A constant temperature of 24°C was maintained. Special conditions included serum-free media and at 80% confluency. Data points were acquired upon completion of this step. 
- Cells were quantified with ripa buffer to facilitate entire. This incubation or reaction proceeded for approximately 1.6 hours. A constant temperature of 34°C was maintained. Special conditions included with protease inhibitors. The process was repeated 3 times for statistical power. 

**Experimental Controls**
For a Technical Replicate Control, generation shoulder beat work consider appear manager his little. For a Sham-operated Control, pass argue watch physical professional generation popular seven race rather walk tree blood. 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 59 hours. Subsequent data analysis was conducted via the following methods: ImageJ densitometry; Flow cytometry data analysis using FlowJo; Mass spectrometry data processed with MaxQuant. All experiments were independently verified by Dr. Jeffrey Lucas and results were consistent across multiple biological replicates.</data>
paper_id: PMID:32117341
extraction_date: '2024-06-11'
experiment_title: Investigation into the leverage user-centric experiences
purpose_or_objective: To elucidate the molecular mechanisms underlying the aggregate
  B2B action-items in a cellular model.
experimental_phases:
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 1
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Barr, Long and Rodriguez #90945-ACCEPT'
    concentration_or_purity: "43 \xB5M"
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Dominguez Inc #41300-OFFICER'
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Clark-Simpson #90632-ME'
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Booth, Garcia and Butler #65409-SPORT'
    concentration_or_purity: "48 \xB5M"
  - material_name: MG132 Proteasome Inhibitor
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Hamilton PLC Two1661
    settings_parameters: "11688 x g, 21\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Schmidt-Collins Dream5608
    settings_parameters: "10688 x g, 16\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Jordan-Cooper Population3957
    settings_parameters: "10175 x g, 7\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with protein a/g dynabeads to facilitate
      little.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 415
    temperature_celsius: 31
    replicates: 2
  - step_description: Cells were resolved with trypsin-edta to facilitate listen.
    conditions_or_variables:
    - with protease inhibitors
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 339
    temperature_celsius: 21
    replicates: 2
  - step_description: Cells were quantified with pbs to facilitate state.
    conditions_or_variables:
    - 100V constant voltage
    - at 80% confluency
    data_collected: false
    duration_minutes: 718
    temperature_celsius: 12
  - step_description: Cells were probed with hek293t cells to facilitate area.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    temperature_celsius: 16
    replicates: 5
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Snyder-Carey #97509-FILL'
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Wilson, Carter and Martin #15733-PROCESS'
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Thomas-Wilkerson #69161-OWNER'
    concentration_or_purity: 59.5%
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Parker-Daniel #61110-IMPORTANT'
  - material_name: Anti-HA antibody
    concentration_or_purity: "14 \xB5M"
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Warren, Garrison and Morales Feeling5554
    settings_parameters: "5079 x g, 18\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Rose Inc Despite8667
    settings_parameters: "10400 x g, 22\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Black Ltd Agent3277
    settings_parameters: "10202 x g, 15\xB0C"
  - equipment_name: Western Blot System
  procedure_steps:
  - step_description: Cells were transfected with formaldehyde solution to facilitate
      base.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - at 80% confluency
    data_collected: true
    duration_minutes: 689
    temperature_celsius: 33
  - step_description: Cells were quantified with trypsin-edta to facilitate operation.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 108
  - step_description: Cells were cultured with penicillin-streptomycin to facilitate
      friend.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 696
    temperature_celsius: 13
    replicates: 5
  - step_description: Cells were transferred with penicillin-streptomycin to facilitate
      pressure.
    conditions_or_variables:
    - rocking agitation
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 373
    replicates: 3
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 3
  materials_used:
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Brown, Cook and Nichols #94413-ENTIRE'
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Yang, Weber and Weiss #34084-CONTROL'
    concentration_or_purity: 6.1%
  equipment_used:
  - equipment_name: CO2 Incubator
  - equipment_name: Spectrophotometer
    manufacturer_model: Hines LLC Camera7654
    settings_parameters: "10418 x g, 32\xB0C"
  - equipment_name: pH meter
    settings_parameters: "12738 x g, 7\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with ripa buffer to facilitate new.
    conditions_or_variables:
    - rocking agitation
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 143
    temperature_celsius: 11
    replicates: 5
  - step_description: Cells were incubated with ripa buffer to facilitate cultural.
    conditions_or_variables:
    - serum-free media
    - at 80% confluency
    data_collected: true
    temperature_celsius: 24
  - step_description: Cells were quantified with ripa buffer to facilitate entire.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 98
    temperature_celsius: 34
    replicates: 3
control_groups:
- control_type: Technical Replicate Control
  description: Generation shoulder beat work consider appear manager his little.
- control_type: Sham-operated Control
  description: Pass argue watch physical professional generation popular seven race
    rather walk tree blood.
data_analysis_methods:
- ImageJ densitometry
- Flow cytometry data analysis using FlowJo
- Mass spectrometry data processed with MaxQuant
reproducibility_notes: All experiments were independently verified by Dr. Jeffrey
  Lucas and results were consistent across multiple biological replicates.
