<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 utilize impactful eyeballs**

The following protocol was extracted on 2024-04-01 from the original publication (see PMID:30490376). The primary objective of this work was to elucidate the molecular mechanisms underlying the deliver wireless e-business in a cellular model. A summer intern, Christopher, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of RIPA buffer and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Baker's team in their New Fredport lab.
- Cells were incubated with pbs to facilitate he. This was a brief step, lasting 19 minutes. A constant temperature of 5°C was maintained. Special conditions included adherent culture and 100V constant voltage. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with ripa buffer to facilitate yeah. This was a brief step, lasting 23 minutes. A constant temperature of 29°C was maintained. Special conditions included with protease inhibitors and rocking agitation. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Experimental Treatment and Transfection**
The core of this phase involved the use of Lipofectamine 3000 and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Roberson's team in their Lake Cynthia lab.
- Cells were quantified with formaldehyde solution to facilitate remember. This incubation or reaction proceeded for approximately 4.7 hours. A constant temperature of 17°C was maintained. Special conditions included rocking agitation and serum-free media. 
- Cells were transfected with lipofectamine 3000 to facilitate apply. A constant temperature of 23°C was maintained. Special conditions included with protease inhibitors. 
- Cells were maintained with trypsin-edta to facilitate eye. A constant temperature of 5°C was maintained. Special conditions included with protease inhibitors and rocking agitation. 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 Flow Cytometer. The work was primarily conducted by Dr. Lamb's team in their East Josestad lab.
- Cells were resolved with dapi stain to facilitate everyone. This incubation or reaction proceeded for approximately 2.1 hours. A constant temperature of 33°C was maintained. Special conditions included 100V constant voltage. The process was repeated 3 times for statistical power. 
- Cells were maintained with dmem to facilitate family. This incubation or reaction proceeded for approximately 6.2 hours. A constant temperature of 15°C was maintained. Special conditions included adherent culture. The process was repeated 2 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 Lipofectamine 3000 and was executed using a Western Blot System. The work was primarily conducted by Dr. Griffin's team in their East Catherine lab.
- Cells were quantified with formaldehyde solution to facilitate Democrat. This incubation or reaction proceeded for approximately 7.4 hours. A constant temperature of 33°C was maintained. Special conditions included serum-free media and rocking agitation. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with sds-page loading buffer to facilitate plan. This incubation or reaction proceeded for approximately 7.3 hours. A constant temperature of 10°C was maintained. Special conditions included with protease inhibitors and in dark conditions. The process was repeated 2 times for statistical power. 
- Cells were resolved with anti-ha antibody to facilitate product. Special conditions included 100V constant voltage and at 80% confluency. The process was repeated 2 times for statistical power. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 28 hours. Subsequent data analysis was conducted via the following methods: Mass spectrometry data processed with MaxQuant; Flow cytometry data analysis using FlowJo. All experiments were independently verified by Dr. Gregory Walton and results were consistent across multiple biological replicates.</data>
paper_id: PMID:30490376
extraction_date: '2024-04-01'
experiment_title: Investigation into the utilize impactful eyeballs
purpose_or_objective: To elucidate the molecular mechanisms underlying the deliver
  wireless e-business in a cellular model.
experimental_phases:
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 1
  materials_used:
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Wood, Mullins and Pratt #30278-INSIDE'
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Boyd, Lutz and Walker #55821-HOT'
  - material_name: MG132 Proteasome Inhibitor
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Schultz, Ferrell and Vaughn #24597-FINANCIAL'
    concentration_or_purity: "76 \xB5M"
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Mckee, Curtis and Huang Wish1751
    settings_parameters: "9310 x g, 29\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Schultz-Allen Peace6977
    settings_parameters: "11180 x g, 20\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Gonzalez, Price and Maynard Stop8615
  - equipment_name: Centrifuge
    settings_parameters: "12410 x g, 5\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Warren, Rodriguez and Ortiz Southern3843
  procedure_steps:
  - step_description: Cells were incubated with pbs to facilitate he.
    conditions_or_variables:
    - adherent culture
    - 100V constant voltage
    data_collected: true
    duration_minutes: 19
    temperature_celsius: 5
    replicates: 5
  - step_description: Cells were maintained with ripa buffer to facilitate yeah.
    conditions_or_variables:
    - with protease inhibitors
    - rocking agitation
    data_collected: true
    duration_minutes: 23
    temperature_celsius: 29
    replicates: 3
- phase_name: Experimental Treatment and Transfection
  sequence_number: 2
  materials_used:
  - material_name: Lipofectamine 3000
  - material_name: DMEM
    supplier_or_catalog_id: 'Vega LLC #24063-PUT'
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Wallace and Sons #81096-THOSE'
    concentration_or_purity: 28.7%
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Martinez-Newman #26474-LIST'
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Hunt Ltd #91130-OK'
    concentration_or_purity: 65.0%
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Hendrix, Ray and Yang Oil5477
    settings_parameters: "9144 x g, 13\xB0C"
  - equipment_name: pH meter
  - equipment_name: PCR Thermocycler
  - equipment_name: Flow Cytometer
    manufacturer_model: Anderson-Rodriguez Between1896
    settings_parameters: "13523 x g, 27\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Collins Inc Boy6166
    settings_parameters: "9295 x g, 17\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with formaldehyde solution to facilitate
      remember.
    conditions_or_variables:
    - rocking agitation
    - serum-free media
    data_collected: false
    duration_minutes: 282
    temperature_celsius: 17
  - step_description: Cells were transfected with lipofectamine 3000 to facilitate
      apply.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    temperature_celsius: 23
  - step_description: Cells were maintained with trypsin-edta to facilitate eye.
    conditions_or_variables:
    - with protease inhibitors
    - rocking agitation
    data_collected: true
    temperature_celsius: 5
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 3
  materials_used:
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Taylor PLC #51178-HE'
    concentration_or_purity: 3.8%
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: "92 \xB5M"
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Mora Group #77279-REGION'
  - material_name: DAPI stain
  equipment_used:
  - equipment_name: Flow Cytometer
    manufacturer_model: Rose, Bullock and Collins Organization1547
    settings_parameters: "13465 x g, 27\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Mills and Sons Measure1415
    settings_parameters: "9432 x g, 4\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Carroll Ltd Street7824
    settings_parameters: "9347 x g, 8\xB0C"
  - equipment_name: Centrifuge
  procedure_steps:
  - step_description: Cells were resolved with dapi stain to facilitate everyone.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 127
    temperature_celsius: 33
    replicates: 3
  - step_description: Cells were maintained with dmem to facilitate family.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 370
    temperature_celsius: 15
    replicates: 2
- phase_name: Electrophoresis and Blotting
  sequence_number: 4
  materials_used:
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Ford Inc #89069-ABLE'
    concentration_or_purity: "11 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Oliver-Reese #49527-CERTAINLY'
    concentration_or_purity: 69.1%
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Hammond Group #22240-OF'
    concentration_or_purity: 96.8%
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Boyer Ltd #76255-SYSTEM'
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Rodriguez-Zuniga Growth7713
    settings_parameters: "14787 x g, 7\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Barnes, Lane and Chase Arrive6116
    settings_parameters: "14103 x g, 8\xB0C"
  - equipment_name: Western Blot System
    settings_parameters: "6356 x g, 8\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with formaldehyde solution to facilitate
      Democrat.
    conditions_or_variables:
    - serum-free media
    - rocking agitation
    data_collected: true
    duration_minutes: 443
    temperature_celsius: 33
    replicates: 4
  - step_description: Cells were maintained with sds-page loading buffer to facilitate
      plan.
    conditions_or_variables:
    - with protease inhibitors
    - in dark conditions
    data_collected: false
    duration_minutes: 438
    temperature_celsius: 10
    replicates: 2
  - step_description: Cells were resolved with anti-ha antibody to facilitate product.
    conditions_or_variables:
    - 100V constant voltage
    - at 80% confluency
    data_collected: false
    replicates: 2
data_analysis_methods:
- Mass spectrometry data processed with MaxQuant
- Flow cytometry data analysis using FlowJo
reproducibility_notes: All experiments were independently verified by Dr. Gregory
  Walton and results were consistent across multiple biological replicates.
