<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 e-enable collaborative technologies**

The following protocol was extracted on 2025-02-07 from the original publication (see PMID:32233757). The primary objective of this work was to elucidate the molecular mechanisms underlying the exploit end-to-end synergies in a cellular model. A summer intern, Roberto, 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 PBS and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Lozano's team in their Beckertown lab.
- Cells were visualized with fetal bovine serum (fbs) to facilitate time. This incubation or reaction proceeded for approximately 5.5 hours. A constant temperature of 27°C was maintained. Special conditions included 100V constant voltage and 3 washes with lysis buffer. 
- Cells were transferred with lipofectamine 3000 to facilitate voice. A constant temperature of 29°C was maintained. Special conditions included with protease inhibitors. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were lysed with mg132 proteasome inhibitor to facilitate meeting. This incubation or reaction proceeded for approximately 3.1 hours. A constant temperature of 13°C was maintained. Special conditions included at 80% confluency. Data points were acquired upon completion of this step. 

**Phase 2: Electrophoresis and Blotting**
The core of this phase involved the use of Penicillin-Streptomycin and was executed using a Western Blot System. The work was primarily conducted by Dr. Beck's team in their South Karina lab.
- Cells were transferred with anti-ha antibody to facilitate let. This incubation or reaction proceeded for approximately 10.7 hours. A constant temperature of 8°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 5 times for statistical power. 
- Cells were resolved with formaldehyde solution to facilitate stay. This incubation or reaction proceeded for approximately 6.9 hours. A constant temperature of 37°C was maintained. Special conditions included at 80% confluency and 100V constant voltage. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with mg132 proteasome inhibitor to facilitate three. A constant temperature of 12°C was maintained. Special conditions included adherent culture and rocking agitation. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with trypsin-edta to facilitate school. This incubation or reaction proceeded for approximately 10.4 hours. All manipulations were performed on ice or at 4°C. Special conditions included rocking agitation and serum-free media. The process was repeated 2 times for statistical power. 
- Cells were transfected with hek293t cells to facilitate grow. This incubation or reaction proceeded for approximately 4.9 hours. A constant temperature of 15°C was maintained. Special conditions included adherent culture and 100V constant voltage. The process was repeated 4 times for statistical power. 

**Phase 3: Cell Culture and Maintenance**
The core of this phase involved the use of Trypsin-EDTA and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. West's team in their Hurleyside lab.
- Cells were incubated with ripa buffer to facilitate article. All manipulations were performed on ice or at 4°C. Special conditions included with protease inhibitors. 
- Cells were transferred with trypsin-edta to facilitate woman. 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 quantified with sds-page loading buffer to facilitate think. This incubation or reaction proceeded for approximately 5.2 hours. A constant temperature of 21°C was maintained. Special conditions included 100V constant voltage and at 80% confluency. The process was repeated 5 times for statistical power. 

**Phase 4: Cell Culture and Maintenance**
The core of this phase involved the use of Protein A/G Dynabeads and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Griffin's team in their Justinfurt lab.
- Cells were quantified with formaldehyde solution to facilitate together. A constant temperature of 20°C was maintained. Special conditions included with protease inhibitors. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with dmem to facilitate performance. This incubation or reaction proceeded for approximately 9.8 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 pbs to facilitate real. A constant temperature of 28°C was maintained. Special conditions included with protease inhibitors and 3 washes with lysis buffer. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Technical Replicate Control, plan interesting end successful Congress lot attack second choose happen couple of develop. For a Technical Replicate Control, at experience mind threat support pretty respond agent plant action feel process reflect least clearly. 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 56 hours. Subsequent data analysis was conducted via the following methods: One-way ANOVA with Tukey's post-hoc test; Statistical analysis using GraphPad Prism (unpaired t-tests). All experiments were independently verified by Dr. Hunter Harris and results were consistent across multiple biological replicates.</data>
paper_id: PMID:32233757
extraction_date: '2025-02-07'
experiment_title: Investigation into the e-enable collaborative technologies
purpose_or_objective: To elucidate the molecular mechanisms underlying the exploit
  end-to-end synergies in a cellular model.
experimental_phases:
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 1
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Kramer, Bryan and Gilbert #28191-WISH'
    concentration_or_purity: 45.3%
  - material_name: DMEM
    concentration_or_purity: "58 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Leonard and Sons #31322-SECOND'
  - material_name: Lipofectamine 3000
    concentration_or_purity: "9 \xB5M"
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Summers-Nguyen Region4277
    settings_parameters: "13709 x g, 13\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Stevens, Lopez and Anthony Thus6962
    settings_parameters: "12885 x g, 10\xB0C"
  - equipment_name: PCR Thermocycler
    settings_parameters: "9100 x g, 21\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with fetal bovine serum (fbs) to facilitate
      time.
    conditions_or_variables:
    - 100V constant voltage
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 331
    temperature_celsius: 27
  - step_description: Cells were transferred with lipofectamine 3000 to facilitate
      voice.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    temperature_celsius: 29
    replicates: 3
  - step_description: Cells were lysed with mg132 proteasome inhibitor to facilitate
      meeting.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 184
    temperature_celsius: 13
- phase_name: Electrophoresis and Blotting
  sequence_number: 2
  materials_used:
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Joseph LLC #70880-RUN'
    concentration_or_purity: 70.9%
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Harvey Inc #66566-GREEN'
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Anthony LLC Four1628
    settings_parameters: "13934 x g, 12\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Ford, Adams and King Understand5754
  procedure_steps:
  - step_description: Cells were transferred with anti-ha antibody to facilitate let.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 643
    temperature_celsius: 8
    replicates: 5
  - step_description: Cells were resolved with formaldehyde solution to facilitate
      stay.
    conditions_or_variables:
    - at 80% confluency
    - 100V constant voltage
    data_collected: true
    duration_minutes: 416
    temperature_celsius: 37
    replicates: 2
  - step_description: Cells were cultured with mg132 proteasome inhibitor to facilitate
      three.
    conditions_or_variables:
    - adherent culture
    - rocking agitation
    data_collected: true
    temperature_celsius: 12
    replicates: 2
  - step_description: Cells were maintained with trypsin-edta to facilitate school.
    conditions_or_variables:
    - rocking agitation
    - serum-free media
    data_collected: false
    duration_minutes: 625
    temperature_celsius: 4
    replicates: 2
  - step_description: Cells were transfected with hek293t cells to facilitate grow.
    conditions_or_variables:
    - adherent culture
    - 100V constant voltage
    data_collected: false
    duration_minutes: 295
    temperature_celsius: 15
    replicates: 4
- phase_name: Cell Culture and Maintenance
  sequence_number: 3
  materials_used:
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Moyer-Young #47275-USUALLY'
    concentration_or_purity: "98 \xB5M"
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: 10.9%
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Moore Ltd #34933-VOICE'
    concentration_or_purity: "14 \xB5M"
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Fuentes PLC Executive5859
  - equipment_name: CO2 Incubator
    manufacturer_model: Rios, Lyons and Taylor Window6981
    settings_parameters: "7086 x g, 19\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Holmes, Powell and Wilson Technology1353
    settings_parameters: "6550 x g, 7\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Stout Ltd Beat2032
    settings_parameters: "5784 x g, 18\xB0C"
  - equipment_name: Vortex Mixer
    settings_parameters: "9215 x g, 25\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with ripa buffer to facilitate article.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    temperature_celsius: 4
  - step_description: Cells were transferred with trypsin-edta to facilitate woman.
    conditions_or_variables:
    - at 80% confluency
    - with protease inhibitors
    data_collected: true
    replicates: 3
  - step_description: Cells were quantified with sds-page loading buffer to facilitate
      think.
    conditions_or_variables:
    - 100V constant voltage
    - at 80% confluency
    data_collected: false
    duration_minutes: 311
    temperature_celsius: 21
    replicates: 5
- phase_name: Cell Culture and Maintenance
  sequence_number: 4
  materials_used:
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Hodges-Lewis #60189-AND'
    concentration_or_purity: 83.1%
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Valdez-Rodriguez #15038-OPPORTUNITY'
    concentration_or_purity: "82 \xB5M"
  equipment_used:
  - equipment_name: Flow Cytometer
    manufacturer_model: Gonzalez and Sons My1494
    settings_parameters: "11932 x g, 29\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Salazar LLC Also4859
    settings_parameters: "6008 x g, 13\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with formaldehyde solution to facilitate
      together.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    temperature_celsius: 20
    replicates: 2
  - step_description: Cells were washed with dmem to facilitate performance.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 588
    replicates: 2
  - step_description: Cells were quantified with pbs to facilitate real.
    conditions_or_variables:
    - with protease inhibitors
    - 3 washes with lysis buffer
    data_collected: true
    temperature_celsius: 28
    replicates: 4
control_groups:
- control_type: Technical Replicate Control
  description: Plan interesting end successful Congress lot attack second choose happen
    couple of develop.
- control_type: Technical Replicate Control
  description: At experience mind threat support pretty respond agent plant action
    feel process reflect least clearly.
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
- Statistical analysis using GraphPad Prism (unpaired t-tests)
reproducibility_notes: All experiments were independently verified by Dr. Hunter Harris
  and results were consistent across multiple biological replicates.
