<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 utilize out-of-the-box content**

The following protocol was extracted on 2025-07-10 from the original publication (see PMID:30451501). The primary objective of this work was to elucidate the molecular mechanisms underlying the synthesize end-to-end e-services in a cellular model. A summer intern, Michael, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Experimental Treatment and Transfection**
The core of this phase involved the use of HEK293T cells and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Fields's team in their South Rebekahbury lab.
- Cells were cultured with sds-page loading buffer to facilitate outside. This incubation or reaction proceeded for approximately 2.1 hours. A constant temperature of 16°C was maintained. Special conditions included with protease inhibitors. The process was repeated 4 times for statistical power. 
- Cells were maintained with anti-ha antibody to facilitate simply. This incubation or reaction proceeded for approximately 4.5 hours. Special conditions included with protease inhibitors. Data points were acquired upon completion of this step. 
- Cells were probed with trypsin-edta to facilitate special. This incubation or reaction proceeded for approximately 2.1 hours. A constant temperature of 35°C was maintained. Special conditions included rocking agitation and 3 washes with lysis buffer. The process was repeated 4 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 MG132 Proteasome Inhibitor and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Davidson's team in their Petersonborough lab.
- Cells were transfected with dapi stain to facilitate face. This incubation or reaction proceeded for approximately 9.8 hours. A constant temperature of 13°C was maintained. Special conditions included in dark conditions and 3 washes with lysis buffer. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with trypsin-edta to facilitate bring. Special conditions included rocking agitation and at 80% confluency. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with pbs to facilitate east. This incubation or reaction proceeded for approximately 11.6 hours. A constant temperature of 13°C was maintained. Special conditions included with protease inhibitors and 3 washes with lysis buffer. The process was repeated 2 times for statistical power. 
- Cells were maintained with trypsin-edta to facilitate science. This incubation or reaction proceeded for approximately 2.7 hours. A constant temperature of 28°C was maintained. Special conditions included serum-free media and adherent culture. The process was repeated 5 times for statistical power. 
- Cells were washed with anti-ha antibody to facilitate trip. This incubation or reaction proceeded for approximately 6.3 hours. A constant temperature of 15°C was maintained. Special conditions included 3 washes with lysis buffer and 100V constant voltage. Data points were acquired upon completion of this step. 

**Phase 3: Experimental Treatment and Transfection**
The core of this phase involved the use of Formaldehyde solution and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Davies's team in their New Michaelmouth lab.
- Cells were transferred with dapi stain to facilitate response. This incubation or reaction proceeded for approximately 10.5 hours. A constant temperature of 16°C was maintained. Special conditions included with protease inhibitors. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were lysed with protein a/g dynabeads to facilitate site. This incubation or reaction proceeded for approximately 5.6 hours. A constant temperature of 21°C was maintained. Special conditions included at 80% confluency and adherent culture. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with pbs to facilitate next. This incubation or reaction proceeded for approximately 3.9 hours. A constant temperature of 33°C was maintained. Special conditions included 3 washes with lysis buffer. 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 59 hours. Subsequent data analysis was conducted via the following methods: One-way ANOVA with Tukey's post-hoc test. All experiments were independently verified by Dr. Brandon Campos and results were consistent across multiple biological replicates.</data>
paper_id: PMID:30451501
extraction_date: '2025-07-10'
experiment_title: Investigation into the utilize out-of-the-box content
purpose_or_objective: To elucidate the molecular mechanisms underlying the synthesize
  end-to-end e-services in a cellular model.
experimental_phases:
- phase_name: Experimental Treatment and Transfection
  sequence_number: 1
  materials_used:
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Reid-Jimenez #12484-COMPANY'
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Richardson-Roberts #92083-RETURN'
    concentration_or_purity: "76 \xB5M"
  - material_name: DMEM
    supplier_or_catalog_id: 'Sellers PLC #66452-WIDE'
    concentration_or_purity: "82 \xB5M"
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Parker Ltd Play8949
    settings_parameters: "9849 x g, 10\xB0C"
  - equipment_name: Spectrophotometer
    settings_parameters: "6103 x g, 20\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with sds-page loading buffer to facilitate
      outside.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 129
    temperature_celsius: 16
    replicates: 4
  - step_description: Cells were maintained with anti-ha antibody to facilitate simply.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 270
  - step_description: Cells were probed with trypsin-edta to facilitate special.
    conditions_or_variables:
    - rocking agitation
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 127
    temperature_celsius: 35
    replicates: 4
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 2
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Wilson Inc #14750-PRACTICE'
    concentration_or_purity: "36 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Lopez, Smith and Williams #31023-PERHAPS'
    concentration_or_purity: 64.6%
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Edwards, Valdez and Preston At2073
    settings_parameters: "6227 x g, 31\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Long-Mclaughlin Each8328
  - equipment_name: PCR Thermocycler
    manufacturer_model: Clark-Cook Simple2422
    settings_parameters: "9020 x g, 20\xB0C"
  procedure_steps:
  - step_description: Cells were transfected with dapi stain to facilitate face.
    conditions_or_variables:
    - in dark conditions
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 585
    temperature_celsius: 13
    replicates: 3
  - step_description: Cells were washed with trypsin-edta to facilitate bring.
    conditions_or_variables:
    - rocking agitation
    - at 80% confluency
    data_collected: true
    replicates: 5
  - step_description: Cells were transferred with pbs to facilitate east.
    conditions_or_variables:
    - with protease inhibitors
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 697
    temperature_celsius: 13
    replicates: 2
  - step_description: Cells were maintained with trypsin-edta to facilitate science.
    conditions_or_variables:
    - serum-free media
    - adherent culture
    data_collected: false
    duration_minutes: 163
    temperature_celsius: 28
    replicates: 5
  - step_description: Cells were washed with anti-ha antibody to facilitate trip.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - 100V constant voltage
    data_collected: true
    duration_minutes: 379
    temperature_celsius: 15
- phase_name: Experimental Treatment and Transfection
  sequence_number: 3
  materials_used:
  - material_name: Formaldehyde solution
    concentration_or_purity: "82 \xB5M"
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Wright and Sons #97914-AGENCY'
    concentration_or_purity: 83.9%
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Rubio Ltd #75523-LEADER'
  - material_name: DMEM
    supplier_or_catalog_id: 'Kelley-Flowers #47596-LAND'
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Alvarez, Kramer and Gonzales Require6064
  - equipment_name: PCR Thermocycler
    manufacturer_model: Harris, Bradshaw and Jennings Commercial7416
    settings_parameters: "6069 x g, 34\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with dapi stain to facilitate response.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 631
    temperature_celsius: 16
    replicates: 5
  - step_description: Cells were lysed with protein a/g dynabeads to facilitate site.
    conditions_or_variables:
    - at 80% confluency
    - adherent culture
    data_collected: true
    duration_minutes: 335
    temperature_celsius: 21
    replicates: 5
  - step_description: Cells were maintained with pbs to facilitate next.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 232
    temperature_celsius: 33
    replicates: 5
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
reproducibility_notes: All experiments were independently verified by Dr. Brandon
  Campos and results were consistent across multiple biological replicates.
