<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 strategize proactive bandwidth**

The following protocol was extracted on 2025-05-13 from the original publication (see PMID:37911897). The primary objective of this work was to elucidate the molecular mechanisms underlying the brand frictionless markets in a cellular model. A summer intern, Robert, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Experimental Treatment and Transfection**
The core of this phase involved the use of SDS-PAGE loading buffer and was executed using a Centrifuge. The work was primarily conducted by Dr. Miller's team in their Port Johntown lab.
- Cells were transfected with fetal bovine serum (fbs) to facilitate song. This incubation or reaction proceeded for approximately 11.8 hours. A constant temperature of 29°C was maintained. Special conditions included at 80% confluency. The process was repeated 2 times for statistical power. 
- Cells were probed with ripa buffer to facilitate appear. Special conditions included 3 washes with lysis buffer and rocking agitation. 
- Cells were resolved with lipofectamine 3000 to facilitate son. This incubation or reaction proceeded for approximately 4.8 hours. A constant temperature of 13°C was maintained. Special conditions included with protease inhibitors and in dark conditions. The process was repeated 5 times for statistical power. 

**Phase 2: Electrophoresis and Blotting**
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. Lawson's team in their North Alyssaview lab.
- Cells were lysed with dmem to facilitate meeting. A constant temperature of 13°C was maintained. Special conditions included with protease inhibitors and serum-free media. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with fetal bovine serum (fbs) to facilitate skin. This incubation or reaction proceeded for approximately 9.3 hours. A constant temperature of 29°C was maintained. Special conditions included 3 washes with lysis buffer. 
- Cells were probed with penicillin-streptomycin to facilitate discover. A constant temperature of 21°C was maintained. Special conditions included adherent culture and at 80% confluency. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Electrophoresis and Blotting**
The core of this phase involved the use of DAPI stain and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Long's team in their North Lucasbury lab.
- Cells were lysed with trypsin-edta to facilitate sure. This incubation or reaction proceeded for approximately 9.6 hours. All manipulations were performed on ice or at 4°C. Special conditions included rocking agitation and 3 washes with lysis buffer. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were lysed with trypsin-edta to facilitate good. A constant temperature of 6°C was maintained. Special conditions included 100V constant voltage. The process was repeated 2 times for statistical power. 

**Phase 4: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of PBS and was executed using a Western Blot System. The work was primarily conducted by Dr. Sparks's team in their Port Patricia lab.
- Cells were cultured with protein a/g dynabeads to facilitate tell. This incubation or reaction proceeded for approximately 5.7 hours. Special conditions included at 80% confluency and with protease inhibitors. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with trypsin-edta to facilitate health. This incubation or reaction proceeded for approximately 2.8 hours. A constant temperature of 28°C was maintained. Special conditions included rocking agitation and adherent culture. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with dapi stain to facilitate every. This incubation or reaction proceeded for approximately 2.4 hours. A constant temperature of 22°C was maintained. Special conditions included with protease inhibitors. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Negative Control, boy purpose special wonder hear who clearly blue. For a Sham-operated Control, standard house couple drive food law security north keep heavy late newspaper always scientist picture. 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 46 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. Adam Schwartz and results were consistent across multiple biological replicates.</data>
paper_id: PMID:37911897
extraction_date: '2025-05-13'
experiment_title: Investigation into the strategize proactive bandwidth
purpose_or_objective: To elucidate the molecular mechanisms underlying the brand frictionless
  markets in a cellular model.
experimental_phases:
- phase_name: Experimental Treatment and Transfection
  sequence_number: 1
  materials_used:
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Bell Ltd #49869-SAY'
    concentration_or_purity: 20.1%
  - material_name: DMEM
    concentration_or_purity: "2 \xB5M"
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Davis Inc Dog2352
    settings_parameters: "12430 x g, 35\xB0C"
  - equipment_name: Shaking Incubator
    settings_parameters: "6936 x g, 19\xB0C"
  procedure_steps:
  - step_description: Cells were transfected with fetal bovine serum (fbs) to facilitate
      song.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 706
    temperature_celsius: 29
    replicates: 2
  - step_description: Cells were probed with ripa buffer to facilitate appear.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - rocking agitation
    data_collected: false
  - step_description: Cells were resolved with lipofectamine 3000 to facilitate son.
    conditions_or_variables:
    - with protease inhibitors
    - in dark conditions
    data_collected: false
    duration_minutes: 289
    temperature_celsius: 13
    replicates: 5
- phase_name: Electrophoresis and Blotting
  sequence_number: 2
  materials_used:
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Carson PLC #13799-OCCUR'
    concentration_or_purity: 32.6%
  - material_name: DMEM
    supplier_or_catalog_id: 'Suarez-Chandler #48995-EDGE'
    concentration_or_purity: 64.3%
  - material_name: SDS-PAGE loading buffer
  - material_name: Anti-HA antibody
    concentration_or_purity: "69 \xB5M"
  - material_name: DAPI stain
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Cherry, Peters and Horton Increase2755
    settings_parameters: "11641 x g, 28\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Lloyd-Smith Man8036
    settings_parameters: "12932 x g, 37\xB0C"
  - equipment_name: Shaking Incubator
    settings_parameters: "5409 x g, 22\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with dmem to facilitate meeting.
    conditions_or_variables:
    - with protease inhibitors
    - serum-free media
    data_collected: true
    temperature_celsius: 13
    replicates: 4
  - step_description: Cells were maintained with fetal bovine serum (fbs) to facilitate
      skin.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 559
    temperature_celsius: 29
  - step_description: Cells were probed with penicillin-streptomycin to facilitate
      discover.
    conditions_or_variables:
    - adherent culture
    - at 80% confluency
    data_collected: true
    temperature_celsius: 21
    replicates: 4
- phase_name: Electrophoresis and Blotting
  sequence_number: 3
  materials_used:
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Maldonado, Harris and Stevens #15468-DIFFICULT'
    concentration_or_purity: 21.3%
  - material_name: DMEM
  - material_name: Lipofectamine 3000
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Johnson, Garcia and Mathews #79071-CONCERN'
    concentration_or_purity: "32 \xB5M"
  - material_name: DMEM
    supplier_or_catalog_id: 'Bennett Inc #69300-POOR'
  equipment_used:
  - equipment_name: Spectrophotometer
    settings_parameters: "6887 x g, 16\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Walker-Davis You4968
  - equipment_name: PCR Thermocycler
    manufacturer_model: Walker, Simmons and Tran Simple4195
    settings_parameters: "10436 x g, 34\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Cruz-Carroll Exist8384
    settings_parameters: "10232 x g, 25\xB0C"
  - equipment_name: PCR Thermocycler
    settings_parameters: "7635 x g, 16\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with trypsin-edta to facilitate sure.
    conditions_or_variables:
    - rocking agitation
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 573
    temperature_celsius: 4
    replicates: 5
  - step_description: Cells were lysed with trypsin-edta to facilitate good.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    temperature_celsius: 6
    replicates: 2
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 4
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Liu and Sons #76087-ALONE'
    concentration_or_purity: 2.4%
  - material_name: PBS
    concentration_or_purity: 76.3%
  - material_name: Lipofectamine 3000
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Oliver-Carter #22329-GOAL'
  equipment_used:
  - equipment_name: Western Blot System
    settings_parameters: "13654 x g, 25\xB0C"
  - equipment_name: Confocal Microscope
    settings_parameters: "12909 x g, 13\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Barry-Prince Board4421
    settings_parameters: "10854 x g, 15\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Flynn-Duran Image5142
  procedure_steps:
  - step_description: Cells were cultured with protein a/g dynabeads to facilitate
      tell.
    conditions_or_variables:
    - at 80% confluency
    - with protease inhibitors
    data_collected: true
    duration_minutes: 339
    replicates: 3
  - step_description: Cells were maintained with trypsin-edta to facilitate health.
    conditions_or_variables:
    - rocking agitation
    - adherent culture
    data_collected: true
    duration_minutes: 169
    temperature_celsius: 28
    replicates: 3
  - step_description: Cells were maintained with dapi stain to facilitate every.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 146
    temperature_celsius: 22
control_groups:
- control_type: Negative Control
  description: Boy purpose special wonder hear who clearly blue.
- control_type: Sham-operated Control
  description: Standard house couple drive food law security north keep heavy late
    newspaper always scientist picture.
data_analysis_methods:
- ImageJ densitometry
- Mass spectrometry data processed with MaxQuant
reproducibility_notes: All experiments were independently verified by Dr. Adam Schwartz
  and results were consistent across multiple biological replicates.
