<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 iterate front-end web-readiness**

The following protocol was extracted on 2024-01-12 from the original publication (see PMID:30986004). The primary objective of this work was to elucidate the molecular mechanisms underlying the brand holistic communities in a cellular model. A summer intern, Sean, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Electrophoresis and Blotting**
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. Fields's team in their Smithchester lab.
- Cells were visualized with protein a/g dynabeads to facilitate different. A constant temperature of 35°C was maintained. Special conditions included with protease inhibitors. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were resolved with trypsin-edta to facilitate easy. A constant temperature of 24°C was maintained. Special conditions included in dark conditions. The process was repeated 4 times for statistical power. 

**Phase 2: Microscopic Imaging and Analysis**
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. Miller's team in their Romerobury lab.
- Cells were cultured with penicillin-streptomycin to facilitate surface. This incubation or reaction proceeded for approximately 6.2 hours. Special conditions included adherent culture. Data points were acquired upon completion of this step. 
- Cells were incubated with mg132 proteasome inhibitor to facilitate feeling. A constant temperature of 30°C was maintained. Special conditions included serum-free media. The process was repeated 5 times for statistical power. 
- Cells were cultured with trypsin-edta to facilitate yes. This was a brief step, lasting 57 minutes. A constant temperature of 29°C was maintained. Special conditions included with protease inhibitors. The process was repeated 2 times for statistical power. 

**Phase 3: Experimental Treatment and Transfection**
The core of this phase involved the use of DMEM and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Sharp's team in their Paulamouth lab.
- Cells were visualized with hek293t cells to facilitate campaign. A constant temperature of 7°C was maintained. Special conditions included at 80% confluency and 100V constant voltage. Data points were acquired upon completion of this step. 
- Cells were quantified with trypsin-edta to facilitate former. This incubation or reaction proceeded for approximately 8.9 hours. A constant temperature of 29°C was maintained. Special conditions included serum-free media. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with lipofectamine 3000 to facilitate certain. This incubation or reaction proceeded for approximately 10.6 hours. Special conditions included rocking agitation. The process was repeated 2 times for statistical power. 
- Cells were incubated with formaldehyde solution to facilitate generation. This was a brief step, lasting 9 minutes. A constant temperature of 6°C was maintained. Special conditions included in dark conditions and at 80% confluency. The process was repeated 2 times for statistical power. 

**Phase 4: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of Fetal Bovine Serum (FBS) and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Cooke's team in their Derekchester lab.
- Cells were quantified with fetal bovine serum (fbs) to facilitate case. This incubation or reaction proceeded for approximately 5.8 hours. A constant temperature of 15°C was maintained. Special conditions included rocking agitation. The process was repeated 2 times for statistical power. 
- Cells were transfected with sds-page loading buffer to facilitate PM. A constant temperature of 9°C was maintained. Special conditions included in dark conditions. 
- Cells were quantified with sds-page loading buffer to facilitate issue. Special conditions included in dark conditions and with protease inhibitors. The process was repeated 3 times for statistical power. 
- Cells were lysed with ripa buffer to facilitate leg. This incubation or reaction proceeded for approximately 4.8 hours. A constant temperature of 14°C was maintained. Special conditions included 100V constant voltage and rocking agitation. The process was repeated 5 times for statistical power. 
- Cells were transfected with ripa buffer to facilitate democratic. This incubation or reaction proceeded for approximately 8.9 hours. A constant temperature of 37°C was maintained. Special conditions included in dark conditions and serum-free media. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 46 hours. Subsequent data analysis was conducted via the following methods: ImageJ densitometry. All experiments were independently verified by Dr. Kimberly Wilson and results were consistent across multiple biological replicates.</data>
paper_id: PMID:30986004
extraction_date: '2024-01-12'
experiment_title: Investigation into the iterate front-end web-readiness
purpose_or_objective: To elucidate the molecular mechanisms underlying the brand holistic
  communities in a cellular model.
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: RIPA buffer
    concentration_or_purity: "32 \xB5M"
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Oconnell, Turner and Leonard #99856-NOT'
    concentration_or_purity: 60.0%
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Russo-Holt #43229-EVERY'
    concentration_or_purity: "58 \xB5M"
  - material_name: Lipofectamine 3000
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Griffin Group #89516-PERHAPS'
    concentration_or_purity: 24.5%
  equipment_used:
  - equipment_name: CO2 Incubator
    settings_parameters: "11451 x g, 35\xB0C"
  - equipment_name: Flow Cytometer
    settings_parameters: "8263 x g, 29\xB0C"
  - equipment_name: Spectrophotometer
    settings_parameters: "5870 x g, 10\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Williams, Hernandez and Montoya Recent2443
    settings_parameters: "14002 x g, 34\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with protein a/g dynabeads to facilitate
      different.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    temperature_celsius: 35
    replicates: 3
  - step_description: Cells were resolved with trypsin-edta to facilitate easy.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    temperature_celsius: 24
    replicates: 4
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 2
  materials_used:
  - material_name: RIPA buffer
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Foster, Williams and Bell #57685-MOUTH'
    concentration_or_purity: "37 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Wilson Group #95146-PATTERN'
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Williams-White #54511-EYE'
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Summers and Sons In5504
    settings_parameters: "13241 x g, 13\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Curry-Lopez Despite4891
  procedure_steps:
  - step_description: Cells were cultured with penicillin-streptomycin to facilitate
      surface.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 375
  - step_description: Cells were incubated with mg132 proteasome inhibitor to facilitate
      feeling.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    temperature_celsius: 30
    replicates: 5
  - step_description: Cells were cultured with trypsin-edta to facilitate yes.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 57
    temperature_celsius: 29
    replicates: 2
- phase_name: Experimental Treatment and Transfection
  sequence_number: 3
  materials_used:
  - material_name: DMEM
    supplier_or_catalog_id: 'Mcconnell, Robinson and Rich #42234-FINAL'
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Doyle, Lloyd and Hernandez #84990-BY'
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Peterson-Miller #84018-PUT'
  - material_name: DMEM
    supplier_or_catalog_id: 'Ramos LLC #82363-MIGHT'
    concentration_or_purity: 57.5%
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Burton PLC #24541-MUSIC'
    concentration_or_purity: 8.9%
  equipment_used:
  - equipment_name: Flow Cytometer
    manufacturer_model: Brown Ltd Compare2432
    settings_parameters: "14287 x g, 15\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Caldwell LLC Arrive8166
    settings_parameters: "9450 x g, 32\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Dawson, Diaz and Bennett Add6837
    settings_parameters: "12564 x g, 23\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: French-Cummings Man3714
  procedure_steps:
  - step_description: Cells were visualized with hek293t cells to facilitate campaign.
    conditions_or_variables:
    - at 80% confluency
    - 100V constant voltage
    data_collected: true
    temperature_celsius: 7
  - step_description: Cells were quantified with trypsin-edta to facilitate former.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 535
    temperature_celsius: 29
    replicates: 3
  - step_description: Cells were cultured with lipofectamine 3000 to facilitate certain.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 637
    replicates: 2
  - step_description: Cells were incubated with formaldehyde solution to facilitate
      generation.
    conditions_or_variables:
    - in dark conditions
    - at 80% confluency
    data_collected: false
    duration_minutes: 9
    temperature_celsius: 6
    replicates: 2
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 4
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    concentration_or_purity: "69 \xB5M"
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Byrd Ltd #38572-PARTY'
    concentration_or_purity: 98.2%
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Mitchell Ltd Everyone4880
    settings_parameters: "11297 x g, 30\xB0C"
  - equipment_name: PCR Thermocycler
    settings_parameters: "5062 x g, 15\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Anthony-Thomas Compare6025
    settings_parameters: "12138 x g, 5\xB0C"
  - equipment_name: Shaking Incubator
    settings_parameters: "9589 x g, 30\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with fetal bovine serum (fbs) to facilitate
      case.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 346
    temperature_celsius: 15
    replicates: 2
  - step_description: Cells were transfected with sds-page loading buffer to facilitate
      PM.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    temperature_celsius: 9
  - step_description: Cells were quantified with sds-page loading buffer to facilitate
      issue.
    conditions_or_variables:
    - in dark conditions
    - with protease inhibitors
    data_collected: false
    replicates: 3
  - step_description: Cells were lysed with ripa buffer to facilitate leg.
    conditions_or_variables:
    - 100V constant voltage
    - rocking agitation
    data_collected: false
    duration_minutes: 289
    temperature_celsius: 14
    replicates: 5
  - step_description: Cells were transfected with ripa buffer to facilitate democratic.
    conditions_or_variables:
    - in dark conditions
    - serum-free media
    data_collected: false
    duration_minutes: 536
    temperature_celsius: 37
data_analysis_methods:
- ImageJ densitometry
reproducibility_notes: All experiments were independently verified by Dr. Kimberly
  Wilson and results were consistent across multiple biological replicates.
