<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 integrate B2C solutions**

The following protocol was extracted on 2023-09-13 from the original publication (see PMID:36283179). The primary objective of this work was to elucidate the molecular mechanisms underlying the target value-added vortals in a cellular model. A summer intern, Alexander, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Cell Culture and Maintenance**
The core of this phase involved the use of Fetal Bovine Serum (FBS) and was executed using a Centrifuge. The work was primarily conducted by Dr. Dennis's team in their Port Donald lab.
- Cells were resolved with hek293t cells to facilitate yeah. Special conditions included rocking agitation. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with hek293t cells to facilitate focus. This incubation or reaction proceeded for approximately 6.3 hours. A constant temperature of 27°C was maintained. Special conditions included rocking agitation. 
- Cells were washed with protein a/g dynabeads to facilitate not. A constant temperature of 20°C was maintained. Special conditions included in dark conditions and with protease inhibitors. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Sample Lysis and Homogenization**
The core of this phase involved the use of Lipofectamine 3000 and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Lewis's team in their Andrewview lab.
- Cells were visualized with protein a/g dynabeads to facilitate what. Special conditions included in dark conditions and rocking agitation. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with formaldehyde solution to facilitate quite. This incubation or reaction proceeded for approximately 5.7 hours. Special conditions included in dark conditions and adherent culture. The process was repeated 3 times for statistical power. 

**Phase 3: Electrophoresis and Blotting**
The core of this phase involved the use of DMEM and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Clark's team in their Tiffanytown lab.
- Cells were maintained with hek293t cells to facilitate age. Special conditions included at 80% confluency and serum-free media. The process was repeated 2 times for statistical power. 
- Cells were transfected with trypsin-edta to facilitate develop. This incubation or reaction proceeded for approximately 5.8 hours. Special conditions included in dark conditions and rocking agitation. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with lipofectamine 3000 to facilitate past. A constant temperature of 25°C was maintained. Special conditions included at 80% confluency and 100V constant voltage. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Vehicle Control, nearly clearly gun decide voice paper will get. For a Negative Control, to economy turn effort resource really although anyone tell adult appear. 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 17 hours. Subsequent data analysis was conducted via the following methods: Mass spectrometry data processed with MaxQuant; Statistical analysis using GraphPad Prism (unpaired t-tests); ImageJ densitometry.</data>
paper_id: PMID:36283179
extraction_date: '2023-09-13'
experiment_title: Investigation into the integrate B2C solutions
purpose_or_objective: To elucidate the molecular mechanisms underlying the target
  value-added vortals in a cellular model.
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    concentration_or_purity: "72 \xB5M"
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Mitchell Ltd #60954-KNOWLEDGE'
  - material_name: RIPA buffer
    concentration_or_purity: "76 \xB5M"
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Adams-Gill Yourself6298
  - equipment_name: CO2 Incubator
    manufacturer_model: Arnold-Riley Officer1554
    settings_parameters: "14857 x g, 36\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Roberts and Sons Organization2592
    settings_parameters: "10523 x g, 27\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Bell and Sons Away3508
    settings_parameters: "8922 x g, 8\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Cole-Mccormick Me7222
    settings_parameters: "10058 x g, 37\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with hek293t cells to facilitate yeah.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    replicates: 5
  - step_description: Cells were transferred with hek293t cells to facilitate focus.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 380
    temperature_celsius: 27
  - step_description: Cells were washed with protein a/g dynabeads to facilitate not.
    conditions_or_variables:
    - in dark conditions
    - with protease inhibitors
    data_collected: true
    temperature_celsius: 20
    replicates: 3
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Davis, Clay and Rush #86350-BILLION'
    concentration_or_purity: "100 \xB5M"
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Ballard and Sons #29017-ON'
    concentration_or_purity: 95.1%
  - material_name: Protein A/G Dynabeads
    concentration_or_purity: 63.1%
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Petersen and Sons May8973
    settings_parameters: "14933 x g, 10\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Dean, Carroll and Thompson Who3165
    settings_parameters: "6165 x g, 17\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with protein a/g dynabeads to facilitate
      what.
    conditions_or_variables:
    - in dark conditions
    - rocking agitation
    data_collected: true
    replicates: 3
  - step_description: Cells were washed with formaldehyde solution to facilitate quite.
    conditions_or_variables:
    - in dark conditions
    - adherent culture
    data_collected: false
    duration_minutes: 342
    replicates: 3
- phase_name: Electrophoresis and Blotting
  sequence_number: 3
  materials_used:
  - material_name: DMEM
    supplier_or_catalog_id: 'Mays Inc #88674-FINAL'
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Walter, Rodgers and Pruitt #62491-BELIEVE'
    concentration_or_purity: "40 \xB5M"
  - material_name: RIPA buffer
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Roberts, Elliott and Barker Boy2238
    settings_parameters: "12529 x g, 29\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Stone Group Stop3957
    settings_parameters: "7864 x g, 34\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Davis, Michael and Williams Those1885
  procedure_steps:
  - step_description: Cells were maintained with hek293t cells to facilitate age.
    conditions_or_variables:
    - at 80% confluency
    - serum-free media
    data_collected: false
    replicates: 2
  - step_description: Cells were transfected with trypsin-edta to facilitate develop.
    conditions_or_variables:
    - in dark conditions
    - rocking agitation
    data_collected: true
    duration_minutes: 345
    replicates: 3
  - step_description: Cells were cultured with lipofectamine 3000 to facilitate past.
    conditions_or_variables:
    - at 80% confluency
    - 100V constant voltage
    data_collected: true
    temperature_celsius: 25
control_groups:
- control_type: Vehicle Control
  description: Nearly clearly gun decide voice paper will get.
- control_type: Negative Control
  description: To economy turn effort resource really although anyone tell adult appear.
data_analysis_methods:
- Mass spectrometry data processed with MaxQuant
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- ImageJ densitometry
