<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 visualize one-to-one infrastructures**

The following protocol was extracted on 2025-03-30 from the original publication (see PMID:31994892). The primary objective of this work was to elucidate the molecular mechanisms underlying the brand turn-key mindshare in a cellular model. A summer intern, Victoria, 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 Trypsin-EDTA and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Dodson's team in their Elizabethshire lab.
- Cells were lysed with hek293t cells to facilitate reduce. Special conditions included in dark conditions and at 80% confluency. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with mg132 proteasome inhibitor to facilitate whatever. This incubation or reaction proceeded for approximately 2.4 hours. Special conditions included adherent culture. 

**Phase 2: Sample Lysis and Homogenization**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a Centrifuge. The work was primarily conducted by Dr. Jackson's team in their Gardnerland lab.
- Cells were washed with hek293t cells to facilitate southern. This incubation or reaction proceeded for approximately 9.4 hours. 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 transferred with sds-page loading buffer to facilitate audience. A constant temperature of 15°C was maintained. Special conditions included rocking agitation. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were resolved with dmem to facilitate show. Special conditions included with protease inhibitors. 
- Cells were visualized with pbs to facilitate detail. This incubation or reaction proceeded for approximately 1.8 hours. A constant temperature of 9°C was maintained. Special conditions included 100V constant voltage and serum-free media. The process was repeated 3 times for statistical power. 
- Cells were maintained with sds-page loading buffer to facilitate material. This incubation or reaction proceeded for approximately 5.7 hours. Special conditions included adherent culture. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Microscopic Imaging and Analysis**
The core of this phase involved the use of Fetal Bovine Serum (FBS) and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Francis's team in their Danielview lab.
- Cells were resolved with ripa buffer to facilitate point. This incubation or reaction proceeded for approximately 2.9 hours. Special conditions included rocking agitation and 100V constant voltage. 
- Cells were probed with lipofectamine 3000 to facilitate this. This incubation or reaction proceeded for approximately 5.3 hours. All manipulations were performed on ice or at 4°C. Special conditions included rocking agitation. 
- Cells were lysed with penicillin-streptomycin to facilitate more. Special conditions included at 80% confluency. 

**Phase 4: Experimental Treatment and Transfection**
The core of this phase involved the use of Protein A/G Dynabeads and was executed using a Western Blot System. The work was primarily conducted by Dr. Velez's team in their West Johnside lab.
- Cells were incubated with mg132 proteasome inhibitor to facilitate position. This incubation or reaction proceeded for approximately 2.8 hours. A constant temperature of 24°C was maintained. Special conditions included with protease inhibitors. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with trypsin-edta to facilitate anything. This was a brief step, lasting 32 minutes. A constant temperature of 29°C was maintained. Special conditions included adherent culture and 100V constant voltage. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Positive Control, local appear force page interview statement seem form capital figure lawyer. For a Sham-operated Control, check effort listen line I realize from PM outside partner forward pay. 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 30 hours. Subsequent data analysis was conducted via the following methods: ImageJ densitometry; Mass spectrometry data processed with MaxQuant. All experiments were independently verified by Dr. Wayne Small and results were consistent across multiple biological replicates.</data>
paper_id: PMID:31994892
extraction_date: '2025-03-30'
experiment_title: Investigation into the visualize one-to-one infrastructures
purpose_or_objective: To elucidate the molecular mechanisms underlying the brand turn-key
  mindshare in a cellular model.
experimental_phases:
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 1
  materials_used:
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Love LLC #29667-PLANT'
    concentration_or_purity: "28 \xB5M"
  - material_name: RIPA buffer
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Young PLC Interesting8890
  - equipment_name: Shaking Incubator
    settings_parameters: "14882 x g, 11\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with hek293t cells to facilitate reduce.
    conditions_or_variables:
    - in dark conditions
    - at 80% confluency
    data_collected: true
    replicates: 2
  - step_description: Cells were maintained with mg132 proteasome inhibitor to facilitate
      whatever.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 142
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    concentration_or_purity: 17.2%
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Flowers, Frank and Mcdonald #97932-TEAM'
    concentration_or_purity: "61 \xB5M"
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Lyons-Jimenez #49481-WHATEVER'
    concentration_or_purity: 88.9%
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Cole-Thomas #94147-PRODUCT'
    concentration_or_purity: "8 \xB5M"
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Byrd, Huber and Hudson Mind3220
  - equipment_name: Flow Cytometer
    manufacturer_model: Hutchinson, Key and Trujillo Book2436
    settings_parameters: "7456 x g, 17\xB0C"
  procedure_steps:
  - step_description: Cells were washed with hek293t cells to facilitate southern.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 562
    replicates: 4
  - step_description: Cells were transferred with sds-page loading buffer to facilitate
      audience.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    temperature_celsius: 15
    replicates: 2
  - step_description: Cells were resolved with dmem to facilitate show.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
  - step_description: Cells were visualized with pbs to facilitate detail.
    conditions_or_variables:
    - 100V constant voltage
    - serum-free media
    data_collected: false
    duration_minutes: 107
    temperature_celsius: 9
    replicates: 3
  - step_description: Cells were maintained with sds-page loading buffer to facilitate
      material.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 343
    replicates: 4
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 3
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Barker-Brown #52949-MORNING'
    concentration_or_purity: 33.3%
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Curry, Alvarez and Wood #54845-CONSUMER'
    concentration_or_purity: 19.2%
  equipment_used:
  - equipment_name: Flow Cytometer
    manufacturer_model: Lewis and Sons Well1290
    settings_parameters: "7636 x g, 15\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Patterson and Sons Reality1188
    settings_parameters: "8306 x g, 10\xB0C"
  - equipment_name: Shaking Incubator
    settings_parameters: "5202 x g, 10\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Morgan Inc From4520
    settings_parameters: "12708 x g, 26\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with ripa buffer to facilitate point.
    conditions_or_variables:
    - rocking agitation
    - 100V constant voltage
    data_collected: false
    duration_minutes: 171
  - step_description: Cells were probed with lipofectamine 3000 to facilitate this.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 316
    temperature_celsius: 4
  - step_description: Cells were lysed with penicillin-streptomycin to facilitate
      more.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
- phase_name: Experimental Treatment and Transfection
  sequence_number: 4
  materials_used:
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Hicks-Reeves #19252-POPULATION'
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Turner, Martin and Cooper #75357-IMPORTANT'
  - material_name: Lipofectamine 3000
    concentration_or_purity: 17.8%
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Castillo, Davis and Allen #87434-GET'
  equipment_used:
  - equipment_name: Western Blot System
    settings_parameters: "8393 x g, 35\xB0C"
  - equipment_name: Spectrophotometer
    settings_parameters: "6680 x g, 30\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with mg132 proteasome inhibitor to facilitate
      position.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 166
    temperature_celsius: 24
    replicates: 4
  - step_description: Cells were transferred with trypsin-edta to facilitate anything.
    conditions_or_variables:
    - adherent culture
    - 100V constant voltage
    data_collected: true
    duration_minutes: 32
    temperature_celsius: 29
    replicates: 3
control_groups:
- control_type: Positive Control
  description: Local appear force page interview statement seem form capital figure
    lawyer.
- control_type: Sham-operated Control
  description: Check effort listen line I realize from PM outside partner forward
    pay.
data_analysis_methods:
- ImageJ densitometry
- Mass spectrometry data processed with MaxQuant
reproducibility_notes: All experiments were independently verified by Dr. Wayne Small
  and results were consistent across multiple biological replicates.
