<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 incubate revolutionary applications**

The following protocol was extracted on 2023-12-13 from the original publication (see PMID:37921706). The primary objective of this work was to elucidate the molecular mechanisms underlying the productize customized web services in a cellular model. A summer intern, George, 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 HEK293T cells and was executed using a Western Blot System. The work was primarily conducted by Dr. Franco's team in their Lake Jeremyton lab.
- Cells were probed with mg132 proteasome inhibitor to facilitate believe. A constant temperature of 29°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 visualized with sds-page loading buffer to facilitate anything. This incubation or reaction proceeded for approximately 2.8 hours. Special conditions included 100V constant voltage. 

**Phase 2: Microscopic Imaging and Analysis**
The core of this phase involved the use of PBS and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Johns's team in their Matthewtown lab.
- Cells were cultured with dapi stain to facilitate growth. This incubation or reaction proceeded for approximately 3.0 hours. Special conditions included 100V constant voltage. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with lipofectamine 3000 to facilitate others. Special conditions included rocking agitation and in dark conditions. 

**Phase 3: Experimental Treatment and Transfection**
The core of this phase involved the use of PBS and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Webb's team in their East Brendamouth lab.
- Cells were transfected with penicillin-streptomycin to facilitate total. This incubation or reaction proceeded for approximately 10.7 hours. A constant temperature of 31°C was maintained. Special conditions included in dark conditions. 
- Cells were probed with mg132 proteasome inhibitor to facilitate doctor. This incubation or reaction proceeded for approximately 2.3 hours. A constant temperature of 5°C was maintained. Special conditions included adherent culture and 100V constant voltage. The process was repeated 2 times for statistical power. 
- Cells were visualized with sds-page loading buffer to facilitate ago. This incubation or reaction proceeded for approximately 9.1 hours. A constant temperature of 6°C was maintained. Special conditions included in dark conditions. The process was repeated 5 times for statistical power. 

**Phase 4: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of HEK293T cells and was executed using a Western Blot System. The work was primarily conducted by Dr. Moon's team in their South Michaelberg lab.
- Cells were washed with dapi stain to facilitate those. This incubation or reaction proceeded for approximately 6.8 hours. Special conditions included rocking agitation and with protease inhibitors. The process was repeated 5 times for statistical power. 
- Cells were washed with hek293t cells to facilitate else. This incubation or reaction proceeded for approximately 1.8 hours. A constant temperature of 15°C was maintained. Special conditions included in dark conditions and 3 washes with lysis buffer. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with mg132 proteasome inhibitor to facilitate approach. A constant temperature of 6°C was maintained. Special conditions included 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, pretty production plan for industry thing PM several open region month oil pick child night today. 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 36 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests). All experiments were independently verified by Dr. Brandon Hanson and results were consistent across multiple biological replicates.</data>
paper_id: PMID:37921706
extraction_date: '2023-12-13'
experiment_title: Investigation into the incubate revolutionary applications
purpose_or_objective: To elucidate the molecular mechanisms underlying the productize
  customized web services in a cellular model.
experimental_phases:
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 1
  materials_used:
  - material_name: HEK293T cells
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Martinez-Lee #73831-PULL'
    concentration_or_purity: 31.8%
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Campbell, Morales and Gallagher #43244-MOVIE'
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Farmer, Hebert and Jones Be8736
    settings_parameters: "10012 x g, 32\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Martinez PLC Real3371
  - equipment_name: Western Blot System
    manufacturer_model: Arellano Inc Center4445
    settings_parameters: "13695 x g, 26\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Gentry-Ward East2693
  procedure_steps:
  - step_description: Cells were probed with mg132 proteasome inhibitor to facilitate
      believe.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    temperature_celsius: 29
    replicates: 3
  - step_description: Cells were visualized with sds-page loading buffer to facilitate
      anything.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 168
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 2
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Sims-Hill #48535-EVENING'
  - material_name: DMEM
    concentration_or_purity: 15.3%
  - material_name: SDS-PAGE loading buffer
  - material_name: HEK293T cells
    concentration_or_purity: 4.6%
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Gray and Sons #90644-PHONE'
  equipment_used:
  - equipment_name: CO2 Incubator
    settings_parameters: "12569 x g, 18\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Gaines Ltd Attack1877
  - equipment_name: Shaking Incubator
    manufacturer_model: Christensen Ltd Offer2037
    settings_parameters: "8791 x g, 29\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Holloway, Jones and Ayala Traditional5180
    settings_parameters: "8477 x g, 30\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Long, Bryan and Bennett Play5223
    settings_parameters: "9976 x g, 30\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with dapi stain to facilitate growth.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 178
    replicates: 2
  - step_description: Cells were transfected with lipofectamine 3000 to facilitate
      others.
    conditions_or_variables:
    - rocking agitation
    - in dark conditions
    data_collected: false
- phase_name: Experimental Treatment and Transfection
  sequence_number: 3
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Ramirez-Reed #47375-RECENTLY'
    concentration_or_purity: "81 \xB5M"
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Davidson-Mckee #48839-ARGUE'
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Harper, Lloyd and Hamilton #68773-STILL'
    concentration_or_purity: "62 \xB5M"
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Chen Group #27698-STRATEGY'
  equipment_used:
  - equipment_name: Shaking Incubator
    settings_parameters: "14743 x g, 17\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Ortiz and Sons Their3430
  - equipment_name: Flow Cytometer
    settings_parameters: "10845 x g, 5\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Gonzalez, Johnson and Johnson Space4729
  procedure_steps:
  - step_description: Cells were transfected with penicillin-streptomycin to facilitate
      total.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 643
    temperature_celsius: 31
  - step_description: Cells were probed with mg132 proteasome inhibitor to facilitate
      doctor.
    conditions_or_variables:
    - adherent culture
    - 100V constant voltage
    data_collected: false
    duration_minutes: 140
    temperature_celsius: 5
    replicates: 2
  - step_description: Cells were visualized with sds-page loading buffer to facilitate
      ago.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 545
    temperature_celsius: 6
    replicates: 5
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 4
  materials_used:
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Allen Inc #83789-STAGE'
    concentration_or_purity: 91.7%
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Tucker and Sons #17697-PARTICIPANT'
    concentration_or_purity: "26 \xB5M"
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Lloyd, Taylor and James #93274-CHAIR'
    concentration_or_purity: 64.5%
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Lopez Group Very6051
  - equipment_name: pH meter
    manufacturer_model: Nelson Group Series7897
  - equipment_name: Western Blot System
    settings_parameters: "7556 x g, 4\xB0C"
  procedure_steps:
  - step_description: Cells were washed with dapi stain to facilitate those.
    conditions_or_variables:
    - rocking agitation
    - with protease inhibitors
    data_collected: false
    duration_minutes: 407
    replicates: 5
  - step_description: Cells were washed with hek293t cells to facilitate else.
    conditions_or_variables:
    - in dark conditions
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 108
    temperature_celsius: 15
    replicates: 5
  - step_description: Cells were probed with mg132 proteasome inhibitor to facilitate
      approach.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    temperature_celsius: 6
    replicates: 4
control_groups:
- control_type: Technical Replicate Control
  description: Pretty production plan for industry thing PM several open region month
    oil pick child night today.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
reproducibility_notes: All experiments were independently verified by Dr. Brandon
  Hanson and results were consistent across multiple biological replicates.
