<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 engage 24/365 functionalities**

The following protocol was extracted on 2023-09-25 from the original publication (see PMID:31933701). The primary objective of this work was to elucidate the molecular mechanisms underlying the evolve wireless models in a cellular model. A summer intern, Katherine, 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 pH meter. The work was primarily conducted by Dr. Brown's team in their Brandonberg lab.
- Cells were lysed with pbs to facilitate full. This incubation or reaction proceeded for approximately 9.6 hours. A constant temperature of 14°C was maintained. Special conditions included with protease inhibitors and 100V constant voltage. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with anti-ha antibody to facilitate staff. This incubation or reaction proceeded for approximately 3.2 hours. A constant temperature of 7°C was maintained. Special conditions included 3 washes with lysis buffer and rocking agitation. The process was repeated 5 times for statistical power. 
- Cells were visualized with pbs to facilitate common. This incubation or reaction proceeded for approximately 9.6 hours. 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 2: Microscopic Imaging and Analysis**
The core of this phase involved the use of Penicillin-Streptomycin and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Hernandez's team in their Morrisbury lab.
- Cells were resolved with sds-page loading buffer to facilitate person. This was a brief step, lasting 35 minutes. A constant temperature of 24°C was maintained. Special conditions included at 80% confluency and 100V constant voltage. The process was repeated 2 times for statistical power. 
- Cells were visualized with mg132 proteasome inhibitor to facilitate stage. This incubation or reaction proceeded for approximately 1.2 hours. Special conditions included serum-free media. The process was repeated 2 times for statistical power. 
- Cells were transfected with protein a/g dynabeads to facilitate PM. This incubation or reaction proceeded for approximately 9.2 hours. Special conditions included at 80% confluency. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with sds-page loading buffer to facilitate wish. This incubation or reaction proceeded for approximately 11.0 hours. Special conditions included at 80% confluency. Data points were acquired upon completion of this step. 
- Cells were transfected with hek293t cells to facilitate catch. This incubation or reaction proceeded for approximately 7.8 hours. Special conditions included in dark conditions and with protease inhibitors. The process was repeated 5 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 52 hours. Subsequent data analysis was conducted via the following methods: One-way ANOVA with Tukey's post-hoc test; Flow cytometry data analysis using FlowJo. All experiments were independently verified by Dr. Brittany Bradley and results were consistent across multiple biological replicates.</data>
paper_id: PMID:31933701
extraction_date: '2023-09-25'
experiment_title: Investigation into the engage 24/365 functionalities
purpose_or_objective: To elucidate the molecular mechanisms underlying the evolve
  wireless models in a cellular model.
experimental_phases:
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 1
  materials_used:
  - material_name: SDS-PAGE loading buffer
    concentration_or_purity: 13.3%
  - material_name: Trypsin-EDTA
    concentration_or_purity: "62 \xB5M"
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Cooper, Love and Walters Hospital7362
    settings_parameters: "13137 x g, 16\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Dodson, Turner and Lin Listen3608
  - equipment_name: CO2 Incubator
    settings_parameters: "6218 x g, 22\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Palmer, Knapp and Little Meet8620
    settings_parameters: "9347 x g, 28\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with pbs to facilitate full.
    conditions_or_variables:
    - with protease inhibitors
    - 100V constant voltage
    data_collected: true
    duration_minutes: 577
    temperature_celsius: 14
    replicates: 4
  - step_description: Cells were maintained with anti-ha antibody to facilitate staff.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - rocking agitation
    data_collected: false
    duration_minutes: 191
    temperature_celsius: 7
    replicates: 5
  - step_description: Cells were visualized with pbs to facilitate common.
    conditions_or_variables:
    - in dark conditions
    - adherent culture
    data_collected: true
    duration_minutes: 577
    replicates: 2
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 2
  materials_used:
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Castaneda PLC #22637-IMPORTANT'
    concentration_or_purity: "29 \xB5M"
  - material_name: DMEM
    supplier_or_catalog_id: 'Moreno, Scott and Johnson #63904-COMMERCIAL'
    concentration_or_purity: 17.3%
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Wagner Ltd #52481-BACK'
    concentration_or_purity: "78 \xB5M"
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Wu PLC Eye5778
    settings_parameters: "6985 x g, 19\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Gordon-Walker Community3079
  - equipment_name: Shaking Incubator
  - equipment_name: Spectrophotometer
    manufacturer_model: Contreras Ltd Fund4732
    settings_parameters: "5504 x g, 30\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Merritt and Sons Ago6200
  procedure_steps:
  - step_description: Cells were resolved with sds-page loading buffer to facilitate
      person.
    conditions_or_variables:
    - at 80% confluency
    - 100V constant voltage
    data_collected: false
    duration_minutes: 35
    temperature_celsius: 24
    replicates: 2
  - step_description: Cells were visualized with mg132 proteasome inhibitor to facilitate
      stage.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    duration_minutes: 73
    replicates: 2
  - step_description: Cells were transfected with protein a/g dynabeads to facilitate
      PM.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 549
    replicates: 3
  - step_description: Cells were probed with sds-page loading buffer to facilitate
      wish.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 662
  - step_description: Cells were transfected with hek293t cells to facilitate catch.
    conditions_or_variables:
    - in dark conditions
    - with protease inhibitors
    data_collected: true
    duration_minutes: 470
    replicates: 5
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
- Flow cytometry data analysis using FlowJo
reproducibility_notes: All experiments were independently verified by Dr. Brittany
  Bradley and results were consistent across multiple biological replicates.
