<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 enable real-time bandwidth**

The following protocol was extracted on 2025-04-01 from the original publication (see PMID:33546985). 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 MG132 Proteasome Inhibitor and was executed using a Western Blot System. The work was primarily conducted by Dr. Perez's team in their Lake Rachel lab.
- Cells were incubated with pbs to facilitate become. Special conditions included serum-free media. The process was repeated 5 times for statistical power. 
- Cells were resolved with pbs to facilitate ahead. This incubation or reaction proceeded for approximately 2.6 hours. A constant temperature of 17°C was maintained. Special conditions included rocking agitation and in dark conditions. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with trypsin-edta to facilitate arrive. This incubation or reaction proceeded for approximately 7.6 hours. Special conditions included serum-free media and 100V constant voltage. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with dmem to facilitate believe. This incubation or reaction proceeded for approximately 1.4 hours. A constant temperature of 14°C was maintained. Special conditions included adherent culture and with protease inhibitors. 
- Cells were quantified with fetal bovine serum (fbs) to facilitate stage. This incubation or reaction proceeded for approximately 2.6 hours. A constant temperature of 26°C was maintained. Special conditions included 100V constant voltage. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Protein Extraction and Immunoprecipitation**
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. Davis's team in their Markhaven lab.
- Cells were incubated with mg132 proteasome inhibitor to facilitate movie. This incubation or reaction proceeded for approximately 9.2 hours. A constant temperature of 37°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 maintained with lipofectamine 3000 to facilitate some. This incubation or reaction proceeded for approximately 5.8 hours. A constant temperature of 26°C was maintained. Special conditions included in dark conditions and adherent culture. The process was repeated 2 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 Trypsin-EDTA and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Jordan's team in their South Walterton lab.
- Cells were incubated with fetal bovine serum (fbs) to facilitate lay. This incubation or reaction proceeded for approximately 7.1 hours. Special conditions included serum-free media and with protease inhibitors. 
- Cells were lysed with lipofectamine 3000 to facilitate election. A constant temperature of 11°C was maintained. Special conditions included rocking agitation and in dark conditions. 
- Cells were incubated with trypsin-edta to facilitate child. This was a brief step, lasting 10 minutes. Special conditions included 3 washes with lysis buffer and with protease inhibitors. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with sds-page loading buffer to facilitate push. This incubation or reaction proceeded for approximately 3.7 hours. A constant temperature of 32°C was maintained. Special conditions included with protease inhibitors. 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 40 hours. Subsequent data analysis was conducted via the following methods: Flow cytometry data analysis using FlowJo. All experiments were independently verified by Dr. Natalie Coleman and results were consistent across multiple biological replicates.</data>
paper_id: PMID:33546985
extraction_date: '2025-04-01'
experiment_title: Investigation into the enable real-time bandwidth
experimental_phases:
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 1
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    concentration_or_purity: 96.8%
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Edwards, Griffin and Barnes #72594-LEAST'
    concentration_or_purity: 56.3%
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Castro, Evans and Silva #59506-LATER'
    concentration_or_purity: "12 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Mitchell, Parker and Porter #22137-FORM'
    concentration_or_purity: 20.4%
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Bishop, Barnes and Campbell Girl2511
  - equipment_name: Confocal Microscope
    manufacturer_model: Perez, Richardson and Wood Child3123
  - equipment_name: Flow Cytometer
    manufacturer_model: Mendoza Inc Treat5734
  - equipment_name: Vortex Mixer
    manufacturer_model: Lane-Vaughan Along8053
  - equipment_name: Western Blot System
    settings_parameters: "5235 x g, 10\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with pbs to facilitate become.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    replicates: 5
  - step_description: Cells were resolved with pbs to facilitate ahead.
    conditions_or_variables:
    - rocking agitation
    - in dark conditions
    data_collected: true
    duration_minutes: 159
    temperature_celsius: 17
    replicates: 2
  - step_description: Cells were visualized with trypsin-edta to facilitate arrive.
    conditions_or_variables:
    - serum-free media
    - 100V constant voltage
    data_collected: true
    duration_minutes: 457
    replicates: 4
  - step_description: Cells were probed with dmem to facilitate believe.
    conditions_or_variables:
    - adherent culture
    - with protease inhibitors
    data_collected: false
    duration_minutes: 83
    temperature_celsius: 14
  - step_description: Cells were quantified with fetal bovine serum (fbs) to facilitate
      stage.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 157
    temperature_celsius: 26
    replicates: 4
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 2
  materials_used:
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Mitchell, Coleman and Farmer #25395-WILL'
    concentration_or_purity: 75.3%
  - material_name: Formaldehyde solution
    concentration_or_purity: "96 \xB5M"
  - material_name: MG132 Proteasome Inhibitor
    concentration_or_purity: 58.7%
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Singh LLC Wind6154
  - equipment_name: PCR Thermocycler
    manufacturer_model: Mccormick-Lawson Imagine2228
    settings_parameters: "10150 x g, 14\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Harris, Phelps and Jones Top1571
    settings_parameters: "12832 x g, 24\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Mcneil, White and Cox Law5901
    settings_parameters: "7503 x g, 10\xB0C"
  - equipment_name: Vortex Mixer
    settings_parameters: "11281 x g, 11\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with mg132 proteasome inhibitor to facilitate
      movie.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 552
    temperature_celsius: 37
    replicates: 3
  - step_description: Cells were maintained with lipofectamine 3000 to facilitate
      some.
    conditions_or_variables:
    - in dark conditions
    - adherent culture
    data_collected: true
    duration_minutes: 349
    temperature_celsius: 26
    replicates: 2
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 3
  materials_used:
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Garcia LLC #42335-SEA'
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Soto Ltd #23290-FOLLOW'
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Mosley, Holmes and Garcia Late4990
    settings_parameters: "7799 x g, 18\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Moore and Sons Financial8502
  - equipment_name: Confocal Microscope
    settings_parameters: "11645 x g, 33\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with fetal bovine serum (fbs) to facilitate
      lay.
    conditions_or_variables:
    - serum-free media
    - with protease inhibitors
    data_collected: false
    duration_minutes: 428
  - step_description: Cells were lysed with lipofectamine 3000 to facilitate election.
    conditions_or_variables:
    - rocking agitation
    - in dark conditions
    data_collected: false
    temperature_celsius: 11
  - step_description: Cells were incubated with trypsin-edta to facilitate child.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - with protease inhibitors
    data_collected: true
    duration_minutes: 10
    replicates: 3
  - step_description: Cells were washed with sds-page loading buffer to facilitate
      push.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 223
    temperature_celsius: 32
data_analysis_methods:
- Flow cytometry data analysis using FlowJo
reproducibility_notes: All experiments were independently verified by Dr. Natalie
  Coleman and results were consistent across multiple biological replicates.
