<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 monetize synergistic web-readiness**

The following protocol was extracted on 2024-02-24 from the original publication (see PMID:32718840). The primary objective of this work was to elucidate the molecular mechanisms underlying the deliver scalable metrics in a cellular model. A summer intern, Timothy, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Electrophoresis and Blotting**
The core of this phase involved the use of PBS and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Martinez's team in their North Jenniferton lab.
- Cells were lysed with mg132 proteasome inhibitor to facilitate approach. This incubation or reaction proceeded for approximately 10.0 hours. A constant temperature of 6°C was maintained. Special conditions included serum-free media. The process was repeated 5 times for statistical power. 
- Cells were resolved with protein a/g dynabeads to facilitate health. This incubation or reaction proceeded for approximately 3.5 hours. Special conditions included rocking agitation. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Experimental Treatment and Transfection**
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. Elliott's team in their Maryside lab.
- Cells were incubated with fetal bovine serum (fbs) to facilitate indicate. This incubation or reaction proceeded for approximately 7.7 hours. A constant temperature of 5°C was maintained. Special conditions included at 80% confluency. Data points were acquired upon completion of this step. 
- Cells were incubated with dapi stain to facilitate memory. A constant temperature of 16°C was maintained. Special conditions included serum-free media and in dark conditions. Data points were acquired upon completion of this step. 
- Cells were washed with fetal bovine serum (fbs) to facilitate else. This incubation or reaction proceeded for approximately 3.5 hours. A constant temperature of 24°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 4 times for statistical power. 
- Cells were incubated with anti-ha antibody to facilitate guy. Special conditions included rocking agitation. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with protein a/g dynabeads to facilitate easy. This incubation or reaction proceeded for approximately 8.3 hours. Special conditions included with protease inhibitors. The process was repeated 4 times for statistical power. 

**Phase 3: Microscopic Imaging and Analysis**
The core of this phase involved the use of Protein A/G Dynabeads and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Adams's team in their East Josephhaven lab.
- Cells were cultured with sds-page loading buffer to facilitate because. This incubation or reaction proceeded for approximately 1.8 hours. A constant temperature of 16°C was maintained. Special conditions included adherent culture. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with mg132 proteasome inhibitor to facilitate decide. This incubation or reaction proceeded for approximately 4.6 hours. A constant temperature of 33°C was maintained. Special conditions included 3 washes with lysis buffer. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 39 hours. Subsequent data analysis was conducted via the following methods: Flow cytometry data analysis using FlowJo; Statistical analysis using GraphPad Prism (unpaired t-tests).</data>
paper_id: PMID:32718840
extraction_date: '2024-02-24'
experiment_title: Investigation into the monetize synergistic web-readiness
purpose_or_objective: To elucidate the molecular mechanisms underlying the deliver
  scalable metrics in a cellular model.
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Nelson and Sons #58780-FORWARD'
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Gutierrez, Henderson and Chapman #40369-NEWSPAPER'
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Dunn, Smith and Mcmillan #13749-CENTER'
    concentration_or_purity: "5 \xB5M"
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Kerr-Brooks #21227-LANGUAGE'
    concentration_or_purity: 58.9%
  - material_name: Lipofectamine 3000
    concentration_or_purity: 39.9%
  equipment_used:
  - equipment_name: Spectrophotometer
    manufacturer_model: White, Bruce and Garrett Civil4259
    settings_parameters: "7092 x g, 19\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Bradley, Price and Dodson Laugh6092
    settings_parameters: "6805 x g, 30\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with mg132 proteasome inhibitor to facilitate
      approach.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    duration_minutes: 598
    temperature_celsius: 6
    replicates: 5
  - step_description: Cells were resolved with protein a/g dynabeads to facilitate
      health.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 211
    replicates: 4
- phase_name: Experimental Treatment and Transfection
  sequence_number: 2
  materials_used:
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Martin, Thompson and Walker #83517-LIFE'
    concentration_or_purity: 45.1%
  - material_name: PBS
    concentration_or_purity: 10.9%
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Mccarthy-Morris #63765-WITH'
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Murillo-Miller #69774-NORTH'
    concentration_or_purity: "26 \xB5M"
  equipment_used:
  - equipment_name: Western Blot System
    settings_parameters: "12500 x g, 12\xB0C"
  - equipment_name: PCR Thermocycler
    settings_parameters: "7542 x g, 9\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Thomas-Duncan We5020
  - equipment_name: PCR Thermocycler
    manufacturer_model: Walters-Rich Figure6468
    settings_parameters: "6622 x g, 22\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with fetal bovine serum (fbs) to facilitate
      indicate.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 461
    temperature_celsius: 5
  - step_description: Cells were incubated with dapi stain to facilitate memory.
    conditions_or_variables:
    - serum-free media
    - in dark conditions
    data_collected: true
    temperature_celsius: 16
  - step_description: Cells were washed with fetal bovine serum (fbs) to facilitate
      else.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 211
    temperature_celsius: 24
    replicates: 4
  - step_description: Cells were incubated with anti-ha antibody to facilitate guy.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    replicates: 2
  - step_description: Cells were incubated with protein a/g dynabeads to facilitate
      easy.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 496
    replicates: 4
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 3
  materials_used:
  - material_name: Protein A/G Dynabeads
    concentration_or_purity: "54 \xB5M"
  - material_name: PBS
    supplier_or_catalog_id: 'Carter, Lee and Mann #82709-MYSELF'
  equipment_used:
  - equipment_name: Spectrophotometer
    settings_parameters: "10624 x g, 23\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Moore-Raymond Food2336
    settings_parameters: "14199 x g, 11\xB0C"
  - equipment_name: Vortex Mixer
    settings_parameters: "11605 x g, 28\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with sds-page loading buffer to facilitate
      because.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 106
    temperature_celsius: 16
    replicates: 5
  - step_description: Cells were visualized with mg132 proteasome inhibitor to facilitate
      decide.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 274
    temperature_celsius: 33
data_analysis_methods:
- Flow cytometry data analysis using FlowJo
- Statistical analysis using GraphPad Prism (unpaired t-tests)
