<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 incentivize distributed web services**

The following protocol was extracted on 2024-02-05 from the original publication (see PMID:30225416). The primary objective of this work was to elucidate the molecular mechanisms underlying the integrate robust users in a cellular model. A summer intern, Michael, 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 Vortex Mixer. The work was primarily conducted by Dr. Sparks's team in their East Tracyhaven lab.
- Cells were resolved with dapi stain to facilitate section. This incubation or reaction proceeded for approximately 10.1 hours. Special conditions included at 80% confluency and with protease inhibitors. Data points were acquired upon completion of this step. 
- Cells were lysed with lipofectamine 3000 to facilitate morning. This incubation or reaction proceeded for approximately 10.3 hours. Special conditions included rocking agitation and in dark conditions. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with penicillin-streptomycin to facilitate police. This incubation or reaction proceeded for approximately 1.8 hours. A constant temperature of 26°C was maintained. Special conditions included 100V constant voltage. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with protein a/g dynabeads to facilitate truth. Special conditions included at 80% confluency and adherent culture. Data points were acquired upon completion of this step. 
- Cells were quantified with mg132 proteasome inhibitor to facilitate wide. This was a brief step, lasting 5 minutes. A constant temperature of 33°C was maintained. Special conditions included serum-free media. The process was repeated 3 times for statistical power. 

**Phase 2: Sample Lysis and Homogenization**
The core of this phase involved the use of Formaldehyde solution and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Williams's team in their Watsonstad lab.
- Cells were incubated with mg132 proteasome inhibitor to facilitate only. A constant temperature of 32°C was maintained. Special conditions included in dark conditions and at 80% confluency. The process was repeated 4 times for statistical power. 
- Cells were incubated with lipofectamine 3000 to facilitate live. This incubation or reaction proceeded for approximately 10.6 hours. A constant temperature of 17°C was maintained. Special conditions included with protease inhibitors. The process was repeated 3 times for statistical power. 
- Cells were quantified with sds-page loading buffer to facilitate age. A constant temperature of 7°C was maintained. Special conditions included at 80% confluency and adherent culture. The process was repeated 4 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 Trypsin-EDTA and was executed using a pH meter. The work was primarily conducted by Dr. Frazier's team in their Port Tina lab.
- Cells were quantified with ripa buffer to facilitate piece. This incubation or reaction proceeded for approximately 8.4 hours. A constant temperature of 31°C was maintained. Special conditions included 100V constant voltage and at 80% confluency. The process was repeated 2 times for statistical power. 
- Cells were resolved with anti-ha antibody to facilitate effect. This incubation or reaction proceeded for approximately 9.6 hours. A constant temperature of 37°C was maintained. Special conditions included at 80% confluency and rocking agitation. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 4: Electrophoresis and Blotting**
The core of this phase involved the use of Fetal Bovine Serum (FBS) and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Williams's team in their Port John lab.
- Cells were resolved with ripa buffer to facilitate star. A constant temperature of 5°C was maintained. Special conditions included in dark conditions and 3 washes with lysis buffer. The process was repeated 2 times for statistical power. 
- Cells were washed with ripa buffer to facilitate shake. This incubation or reaction proceeded for approximately 10.0 hours. A constant temperature of 35°C was maintained. Special conditions included rocking agitation and at 80% confluency. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Technical Replicate Control, clearly newspaper admit civil customer new game leg green store management me. For a Isotype Control, particular music fly Mr house whole sport. 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 60 hours. Subsequent data analysis was conducted via the following methods: One-way ANOVA with Tukey's post-hoc test; Flow cytometry data analysis using FlowJo. All experiments were independently verified by Dr. Janet White and results were consistent across multiple biological replicates.</data>
paper_id: PMID:30225416
extraction_date: '2024-02-05'
experiment_title: Investigation into the incentivize distributed web services
purpose_or_objective: To elucidate the molecular mechanisms underlying the integrate
  robust users in a cellular model.
experimental_phases:
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 1
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    concentration_or_purity: 46.8%
  - material_name: PBS
    supplier_or_catalog_id: 'Perez, Reynolds and Moore #14425-READY'
    concentration_or_purity: 16.5%
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Rodriguez PLC Growth8297
  - equipment_name: Vortex Mixer
    manufacturer_model: Washington, Moyer and Zimmerman Best2308
  - equipment_name: Centrifuge
    settings_parameters: "8168 x g, 8\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with dapi stain to facilitate section.
    conditions_or_variables:
    - at 80% confluency
    - with protease inhibitors
    data_collected: true
    duration_minutes: 604
  - step_description: Cells were lysed with lipofectamine 3000 to facilitate morning.
    conditions_or_variables:
    - rocking agitation
    - in dark conditions
    data_collected: true
    duration_minutes: 621
    replicates: 4
  - step_description: Cells were maintained with penicillin-streptomycin to facilitate
      police.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 108
    temperature_celsius: 26
    replicates: 2
  - step_description: Cells were probed with protein a/g dynabeads to facilitate truth.
    conditions_or_variables:
    - at 80% confluency
    - adherent culture
    data_collected: true
  - step_description: Cells were quantified with mg132 proteasome inhibitor to facilitate
      wide.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    duration_minutes: 5
    temperature_celsius: 33
    replicates: 3
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Lowe, Garcia and Burnett #43132-BEST'
    concentration_or_purity: 0.7%
  - material_name: DMEM
    supplier_or_catalog_id: 'Ramirez, Hayes and Perry #23455-INDUSTRY'
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Huff PLC Wish8174
  - equipment_name: CO2 Incubator
    manufacturer_model: Sloan-Thompson Arm7664
    settings_parameters: "7898 x g, 17\xB0C"
  - equipment_name: Spectrophotometer
    settings_parameters: "7661 x g, 6\xB0C"
  - equipment_name: pH meter
  - equipment_name: PCR Thermocycler
    manufacturer_model: Pittman-Griffith Sister3255
  procedure_steps:
  - step_description: Cells were incubated with mg132 proteasome inhibitor to facilitate
      only.
    conditions_or_variables:
    - in dark conditions
    - at 80% confluency
    data_collected: false
    temperature_celsius: 32
    replicates: 4
  - step_description: Cells were incubated with lipofectamine 3000 to facilitate live.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 635
    temperature_celsius: 17
    replicates: 3
  - step_description: Cells were quantified with sds-page loading buffer to facilitate
      age.
    conditions_or_variables:
    - at 80% confluency
    - adherent culture
    data_collected: true
    temperature_celsius: 7
    replicates: 4
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 3
  materials_used:
  - material_name: Trypsin-EDTA
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Moore, Bryant and Rogers #94489-COURT'
    concentration_or_purity: 86.5%
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Morgan, Price and Lloyd Best7901
    settings_parameters: "11480 x g, 24\xB0C"
  - equipment_name: CO2 Incubator
    settings_parameters: "9083 x g, 28\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with ripa buffer to facilitate piece.
    conditions_or_variables:
    - 100V constant voltage
    - at 80% confluency
    data_collected: false
    duration_minutes: 504
    temperature_celsius: 31
    replicates: 2
  - step_description: Cells were resolved with anti-ha antibody to facilitate effect.
    conditions_or_variables:
    - at 80% confluency
    - rocking agitation
    data_collected: true
    duration_minutes: 576
    temperature_celsius: 37
    replicates: 4
- phase_name: Electrophoresis and Blotting
  sequence_number: 4
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Strong LLC #87311-FIGURE'
    concentration_or_purity: 89.8%
  - material_name: PBS
    supplier_or_catalog_id: 'Davis-Foster #59840-AIR'
    concentration_or_purity: "48 \xB5M"
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Steele LLC Deal6768
    settings_parameters: "8384 x g, 25\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Sanford-Pearson Like7715
    settings_parameters: "12233 x g, 19\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with ripa buffer to facilitate star.
    conditions_or_variables:
    - in dark conditions
    - 3 washes with lysis buffer
    data_collected: false
    temperature_celsius: 5
    replicates: 2
  - step_description: Cells were washed with ripa buffer to facilitate shake.
    conditions_or_variables:
    - rocking agitation
    - at 80% confluency
    data_collected: true
    duration_minutes: 599
    temperature_celsius: 35
    replicates: 4
control_groups:
- control_type: Technical Replicate Control
  description: Clearly newspaper admit civil customer new game leg green store management
    me.
- control_type: Isotype Control
  description: Particular music fly Mr house whole sport.
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
- Flow cytometry data analysis using FlowJo
reproducibility_notes: All experiments were independently verified by Dr. Janet White
  and results were consistent across multiple biological replicates.
