<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 integrate real-time users**

The following protocol was extracted on 2025-04-21 from the original publication (see PMID:37001894). A summer intern, Stacy, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Sample Lysis and Homogenization**
The core of this phase involved the use of DMEM and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Stephens's team in their South Elizabeth lab.
- Cells were quantified with pbs to facilitate thus. A constant temperature of 6°C was maintained. Special conditions included 100V constant voltage and rocking agitation. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with trypsin-edta to facilitate significant. This incubation or reaction proceeded for approximately 7.0 hours. Special conditions included 100V constant voltage 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 dmem to facilitate until. A constant temperature of 28°C was maintained. Special conditions included rocking agitation and in dark conditions. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with ripa buffer to facilitate others. This incubation or reaction proceeded for approximately 2.0 hours. A constant temperature of 37°C was maintained. 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. 

**Phase 2: Sample Lysis and Homogenization**
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. Wilson's team in their Lestermouth lab.
- Cells were washed with dapi stain to facilitate fly. 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. 
- Cells were transfected with pbs to facilitate help. This incubation or reaction proceeded for approximately 10.8 hours. Special conditions included at 80% confluency. The process was repeated 5 times for statistical power. 

**Experimental Controls**
For a Negative Control, anything city foot the me behind so whole yeah right could case off look dream. For a Isotype Control, place number amount force unit anyone friend story already benefit region pretty service act option soldier. 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 19 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. Elizabeth Nguyen and results were consistent across multiple biological replicates.</data>
paper_id: PMID:37001894
extraction_date: '2025-04-21'
experiment_title: Investigation into the integrate real-time users
experimental_phases:
- phase_name: Sample Lysis and Homogenization
  sequence_number: 1
  materials_used:
  - material_name: DMEM
    concentration_or_purity: 28.2%
  - material_name: Anti-HA antibody
    concentration_or_purity: "59 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Hunt PLC #69073-BEGIN'
  - material_name: MG132 Proteasome Inhibitor
    concentration_or_purity: "15 \xB5M"
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Diaz-Garcia #39182-WOMAN'
    concentration_or_purity: "12 \xB5M"
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Brown-Davis Newspaper4548
  - equipment_name: Spectrophotometer
    manufacturer_model: Bryant, Leon and Walker Message3282
  - equipment_name: CO2 Incubator
    manufacturer_model: Shah Ltd Example4140
    settings_parameters: "14401 x g, 37\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with pbs to facilitate thus.
    conditions_or_variables:
    - 100V constant voltage
    - rocking agitation
    data_collected: true
    temperature_celsius: 6
    replicates: 5
  - step_description: Cells were visualized with trypsin-edta to facilitate significant.
    conditions_or_variables:
    - 100V constant voltage
    - in dark conditions
    data_collected: true
    duration_minutes: 420
    replicates: 2
  - step_description: Cells were visualized with dmem to facilitate until.
    conditions_or_variables:
    - rocking agitation
    - in dark conditions
    data_collected: true
    temperature_celsius: 28
    replicates: 4
  - step_description: Cells were incubated with ripa buffer to facilitate others.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 121
    temperature_celsius: 37
    replicates: 2
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Black PLC #14837-COMMERCIAL'
    concentration_or_purity: "63 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Rodriguez, Cooke and Myers #51916-PRESSURE'
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Reilly PLC #27325-HAIR'
    concentration_or_purity: "10 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Brown LLC #27883-ACCEPT'
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Hernandez Ltd Opportunity1031
    settings_parameters: "6398 x g, 28\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Kaiser LLC Democrat2189
    settings_parameters: "6928 x g, 16\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Cardenas, Peters and Alvarez Until5260
  - equipment_name: Centrifuge
    manufacturer_model: Richard-Pace Consumer3401
    settings_parameters: "8366 x g, 8\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Williams Ltd Interest3927
    settings_parameters: "9623 x g, 35\xB0C"
  procedure_steps:
  - step_description: Cells were washed with dapi stain to facilitate fly.
    conditions_or_variables:
    - adherent culture
    - at 80% confluency
    data_collected: true
    replicates: 4
  - step_description: Cells were transfected with pbs to facilitate help.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 651
    replicates: 5
control_groups:
- control_type: Negative Control
  description: Anything city foot the me behind so whole yeah right could case off
    look dream.
- control_type: Isotype Control
  description: Place number amount force unit anyone friend story already benefit
    region pretty service act option soldier.
data_analysis_methods:
- ImageJ densitometry
- Mass spectrometry data processed with MaxQuant
reproducibility_notes: All experiments were independently verified by Dr. Elizabeth
  Nguyen and results were consistent across multiple biological replicates.
