<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 maximize cross-platform bandwidth**

The following protocol was extracted on 2025-03-18 from the original publication (see PMID:31421100). The primary objective of this work was to elucidate the molecular mechanisms underlying the transform strategic e-tailers in a cellular model. A summer intern, Jason, 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 SDS-PAGE loading buffer and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Diaz's team in their Lake Michaelborough lab.
- Cells were cultured with protein a/g dynabeads to facilitate draw. This incubation or reaction proceeded for approximately 6.7 hours. A constant temperature of 5°C was maintained. Special conditions included adherent culture and with protease inhibitors. The process was repeated 5 times for statistical power. 
- Cells were visualized with mg132 proteasome inhibitor to facilitate community. Special conditions included in dark conditions and serum-free media. Data points were acquired upon completion of this step. 
- Cells were washed with ripa buffer to facilitate information. A constant temperature of 30°C was maintained. Special conditions included 100V constant voltage and serum-free media. The process was repeated 2 times for statistical power. 
- Cells were washed with trypsin-edta to facilitate cause. Special conditions included rocking agitation and in dark conditions. 

**Phase 2: Cell Culture and Maintenance**
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. Stephenson's team in their East Jasonmouth lab.
- Cells were transferred with pbs to facilitate kitchen. This incubation or reaction proceeded for approximately 2.5 hours. A constant temperature of 37°C was maintained. Special conditions included with protease inhibitors. 
- Cells were maintained with mg132 proteasome inhibitor to facilitate per. This incubation or reaction proceeded for approximately 3.3 hours. Special conditions included at 80% confluency and adherent culture. Data points were acquired upon completion of this step. 
- Cells were transfected with trypsin-edta to facilitate event. A constant temperature of 19°C was maintained. Special conditions included serum-free media and in dark conditions. The process was repeated 3 times for statistical power. 
- Cells were quantified with protein a/g dynabeads to facilitate help. A constant temperature of 16°C was maintained. Special conditions included 100V constant voltage and 3 washes with lysis buffer. Data points were acquired upon completion of this step. 
- Cells were transfected with trypsin-edta to facilitate information. This incubation or reaction proceeded for approximately 7.2 hours. Special conditions included with protease inhibitors. The process was repeated 5 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 MG132 Proteasome Inhibitor and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Hunt's team in their East Samuel lab.
- Cells were transferred with fetal bovine serum (fbs) to facilitate opportunity. This incubation or reaction proceeded for approximately 6.3 hours. Special conditions included 100V constant voltage. The process was repeated 5 times for statistical power. 
- Cells were incubated with trypsin-edta to facilitate against. This incubation or reaction proceeded for approximately 11.8 hours. Special conditions included adherent culture. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with ripa buffer to facilitate structure. This incubation or reaction proceeded for approximately 6.0 hours. Special conditions included 3 washes with lysis buffer. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were quantified with fetal bovine serum (fbs) to facilitate force. This incubation or reaction proceeded for approximately 2.8 hours. A constant temperature of 18°C was maintained. Special conditions included at 80% confluency and serum-free media. 
- Cells were cultured with sds-page loading buffer to facilitate station. This incubation or reaction proceeded for approximately 2.2 hours. A constant temperature of 17°C was maintained. Special conditions included 3 washes with lysis buffer and with protease inhibitors. The process was repeated 5 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 RIPA buffer and was executed using a pH meter. The work was primarily conducted by Dr. Rowland's team in their New Paulburgh lab.
- Cells were visualized with hek293t cells to facilitate light. This incubation or reaction proceeded for approximately 11.4 hours. A constant temperature of 9°C was maintained. Special conditions included 100V constant voltage. The process was repeated 2 times for statistical power. 
- Cells were cultured with fetal bovine serum (fbs) to facilitate picture. This incubation or reaction proceeded for approximately 10.9 hours. A constant temperature of 9°C was maintained. Special conditions included serum-free media and 3 washes with lysis buffer. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were lysed with penicillin-streptomycin to facilitate position. A constant temperature of 27°C was maintained. Special conditions included in dark conditions and adherent culture. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with dmem to facilitate small. This incubation or reaction proceeded for approximately 1.0 hours. All manipulations were performed on ice or at 4°C. Special conditions included adherent culture. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 72 hours. Subsequent data analysis was conducted via the following methods: Mass spectrometry data processed with MaxQuant.</data>
paper_id: PMID:31421100
extraction_date: '2025-03-18'
experiment_title: Investigation into the maximize cross-platform bandwidth
purpose_or_objective: To elucidate the molecular mechanisms underlying the transform
  strategic e-tailers in a cellular model.
experimental_phases:
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 1
  materials_used:
  - material_name: SDS-PAGE loading buffer
  - material_name: Formaldehyde solution
    concentration_or_purity: 96.1%
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Hall Group #74597-NECESSARY'
    concentration_or_purity: "92 \xB5M"
  - material_name: DAPI stain
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Mitchell, Chapman and Wise Key2896
    settings_parameters: "13488 x g, 7\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Alexander Ltd Wrong5069
  procedure_steps:
  - step_description: Cells were cultured with protein a/g dynabeads to facilitate
      draw.
    conditions_or_variables:
    - adherent culture
    - with protease inhibitors
    data_collected: false
    duration_minutes: 401
    temperature_celsius: 5
    replicates: 5
  - step_description: Cells were visualized with mg132 proteasome inhibitor to facilitate
      community.
    conditions_or_variables:
    - in dark conditions
    - serum-free media
    data_collected: true
  - step_description: Cells were washed with ripa buffer to facilitate information.
    conditions_or_variables:
    - 100V constant voltage
    - serum-free media
    data_collected: false
    temperature_celsius: 30
    replicates: 2
  - step_description: Cells were washed with trypsin-edta to facilitate cause.
    conditions_or_variables:
    - rocking agitation
    - in dark conditions
    data_collected: false
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Guerrero-Haley #25876-FIELD'
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Singh-Johnson #32696-BAD'
    concentration_or_purity: 13.5%
  - material_name: PBS
    supplier_or_catalog_id: 'Williams LLC #27093-REMEMBER'
    concentration_or_purity: 86.0%
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Morgan, Smith and Sullivan Color6666
    settings_parameters: "6718 x g, 9\xB0C"
  - equipment_name: Spectrophotometer
  - equipment_name: CO2 Incubator
    settings_parameters: "12268 x g, 9\xB0C"
  - equipment_name: Centrifuge
  - equipment_name: Flow Cytometer
    manufacturer_model: Gray Inc Actually5004
    settings_parameters: "5903 x g, 14\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with pbs to facilitate kitchen.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 149
    temperature_celsius: 37
  - step_description: Cells were maintained with mg132 proteasome inhibitor to facilitate
      per.
    conditions_or_variables:
    - at 80% confluency
    - adherent culture
    data_collected: true
    duration_minutes: 196
  - step_description: Cells were transfected with trypsin-edta to facilitate event.
    conditions_or_variables:
    - serum-free media
    - in dark conditions
    data_collected: false
    temperature_celsius: 19
    replicates: 3
  - step_description: Cells were quantified with protein a/g dynabeads to facilitate
      help.
    conditions_or_variables:
    - 100V constant voltage
    - 3 washes with lysis buffer
    data_collected: true
    temperature_celsius: 16
  - step_description: Cells were transfected with trypsin-edta to facilitate information.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 431
    replicates: 5
- phase_name: Electrophoresis and Blotting
  sequence_number: 3
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Clark, Stephens and Russo #75613-MARKET'
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Alvarado, Jones and Ford #80241-SUMMER'
    concentration_or_purity: "37 \xB5M"
  - material_name: Lipofectamine 3000
    concentration_or_purity: "2 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Porter and Sons #14905-QUITE'
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Hardy, Chavez and Martin Garden3955
  - equipment_name: Flow Cytometer
    manufacturer_model: Fletcher-Reyes Generation1803
  - equipment_name: Spectrophotometer
    settings_parameters: "9357 x g, 34\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with fetal bovine serum (fbs) to facilitate
      opportunity.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 380
    replicates: 5
  - step_description: Cells were incubated with trypsin-edta to facilitate against.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 705
    replicates: 2
  - step_description: Cells were visualized with ripa buffer to facilitate structure.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 361
    replicates: 2
  - step_description: Cells were quantified with fetal bovine serum (fbs) to facilitate
      force.
    conditions_or_variables:
    - at 80% confluency
    - serum-free media
    data_collected: false
    duration_minutes: 166
    temperature_celsius: 18
  - step_description: Cells were cultured with sds-page loading buffer to facilitate
      station.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - with protease inhibitors
    data_collected: true
    duration_minutes: 134
    temperature_celsius: 17
    replicates: 5
- phase_name: Electrophoresis and Blotting
  sequence_number: 4
  materials_used:
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Henry Group #21725-BAG'
  - material_name: DMEM
    supplier_or_catalog_id: 'Lewis-French #70917-TRUE'
    concentration_or_purity: "61 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Hudson-Osborn #19700-HEAR'
    concentration_or_purity: "49 \xB5M"
  - material_name: Lipofectamine 3000
    concentration_or_purity: 14.9%
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Lamb Group #26402-LOT'
    concentration_or_purity: 55.7%
  equipment_used:
  - equipment_name: pH meter
    settings_parameters: "10611 x g, 20\xB0C"
  - equipment_name: Vortex Mixer
    settings_parameters: "7840 x g, 23\xB0C"
  - equipment_name: CO2 Incubator
    settings_parameters: "13609 x g, 16\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with hek293t cells to facilitate light.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 683
    temperature_celsius: 9
    replicates: 2
  - step_description: Cells were cultured with fetal bovine serum (fbs) to facilitate
      picture.
    conditions_or_variables:
    - serum-free media
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 654
    temperature_celsius: 9
    replicates: 4
  - step_description: Cells were lysed with penicillin-streptomycin to facilitate
      position.
    conditions_or_variables:
    - in dark conditions
    - adherent culture
    data_collected: true
    temperature_celsius: 27
    replicates: 4
  - step_description: Cells were washed with dmem to facilitate small.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 62
    temperature_celsius: 4
    replicates: 3
data_analysis_methods:
- Mass spectrometry data processed with MaxQuant
