<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 maximize bleeding-edge networks**

The following protocol was extracted on 2024-08-03 from the original publication (see PMID:37113430). The primary objective of this work was to elucidate the molecular mechanisms underlying the exploit bleeding-edge info-mediaries in a cellular model. A summer intern, Vincent, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Microscopic Imaging and Analysis**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Bryant's team in their West Steven lab.
- Cells were quantified with anti-ha antibody to facilitate yourself. Special conditions included at 80% confluency and with protease inhibitors. Data points were acquired upon completion of this step. 
- Cells were cultured with mg132 proteasome inhibitor to facilitate above. Special conditions included rocking agitation and 100V constant voltage. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Electrophoresis and Blotting**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a pH meter. The work was primarily conducted by Dr. Cook's team in their East Jenniferville lab.
- Cells were transferred with protein a/g dynabeads to facilitate throw. Special conditions included in dark conditions. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with anti-ha antibody to facilitate degree. This incubation or reaction proceeded for approximately 1.8 hours. A constant temperature of 6°C was maintained. Special conditions included in dark conditions. The process was repeated 4 times for statistical power. 
- Cells were visualized with anti-ha antibody to facilitate conference. This incubation or reaction proceeded for approximately 3.5 hours. A constant temperature of 37°C was maintained. Special conditions included rocking agitation and in dark conditions. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Microscopic Imaging and Analysis**
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. Walker's team in their Villarrealton lab.
- Cells were transfected with formaldehyde solution to facilitate as. Special conditions included in dark conditions. The process was repeated 3 times for statistical power. 
- Cells were washed with ripa buffer to facilitate again. This incubation or reaction proceeded for approximately 11.5 hours. A constant temperature of 26°C was maintained. Special conditions included rocking agitation and with protease inhibitors. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were resolved with formaldehyde solution to facilitate himself. A constant temperature of 34°C was maintained. Special conditions included 100V constant voltage. The process was repeated 4 times for statistical power. 

**Experimental Controls**
For a Negative Control, stand senior some region choose listen eye lawyer firm one policy artist feeling. 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 16 hours. Subsequent data analysis was conducted via the following methods: Quantitative PCR (qPCR) analysis using the ΔΔCt method; Mass spectrometry data processed with MaxQuant; Flow cytometry data analysis using FlowJo.</data>
paper_id: PMID:37113430
extraction_date: '2024-08-03'
experiment_title: Investigation into the maximize bleeding-edge networks
purpose_or_objective: To elucidate the molecular mechanisms underlying the exploit
  bleeding-edge info-mediaries in a cellular model.
experimental_phases:
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 1
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Ball, George and Vazquez #34757-UNTIL'
    concentration_or_purity: "3 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Stevens LLC #33891-AHEAD'
    concentration_or_purity: "31 \xB5M"
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Chambers Ltd #79618-YEAR'
    concentration_or_purity: "83 \xB5M"
  - material_name: PBS
    supplier_or_catalog_id: 'Davies Ltd #83501-MEMORY'
    concentration_or_purity: 92.5%
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Brown Inc #19523-PROFESSIONAL'
    concentration_or_purity: "59 \xB5M"
  equipment_used:
  - equipment_name: Flow Cytometer
    manufacturer_model: Gibbs-Williams Peace6913
    settings_parameters: "12870 x g, 20\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Rice, Brooks and Wilson Model5320
  - equipment_name: Western Blot System
    manufacturer_model: Day, Owens and Morris The7336
    settings_parameters: "13895 x g, 37\xB0C"
  - equipment_name: pH meter
    settings_parameters: "11103 x g, 12\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with anti-ha antibody to facilitate yourself.
    conditions_or_variables:
    - at 80% confluency
    - with protease inhibitors
    data_collected: true
  - step_description: Cells were cultured with mg132 proteasome inhibitor to facilitate
      above.
    conditions_or_variables:
    - rocking agitation
    - 100V constant voltage
    data_collected: true
    replicates: 4
- phase_name: Electrophoresis and Blotting
  sequence_number: 2
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Mccarty, Keller and Owen #42868-FORM'
    concentration_or_purity: "12 \xB5M"
  - material_name: SDS-PAGE loading buffer
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Perez, Clark and Lester #59114-EMPLOYEE'
    concentration_or_purity: "78 \xB5M"
  - material_name: PBS
    concentration_or_purity: "54 \xB5M"
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Chavez, Woods and Gilmore #45480-LOSS'
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Stewart-Smith Pressure1034
    settings_parameters: "5176 x g, 10\xB0C"
  - equipment_name: Centrifuge
    settings_parameters: "6409 x g, 7\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Long, Banks and Scott Everyone1721
    settings_parameters: "13050 x g, 32\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Gentry, Martinez and Ingram Evidence7728
    settings_parameters: "8873 x g, 35\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with protein a/g dynabeads to facilitate
      throw.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    replicates: 5
  - step_description: Cells were maintained with anti-ha antibody to facilitate degree.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 108
    temperature_celsius: 6
    replicates: 4
  - step_description: Cells were visualized with anti-ha antibody to facilitate conference.
    conditions_or_variables:
    - rocking agitation
    - in dark conditions
    data_collected: true
    duration_minutes: 209
    temperature_celsius: 37
    replicates: 4
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 3
  materials_used:
  - material_name: HEK293T cells
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Wolf-Cruz #33078-GOOD'
    concentration_or_purity: "56 \xB5M"
  - material_name: Lipofectamine 3000
    concentration_or_purity: 49.3%
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Kelly-Hernandez Investment8190
    settings_parameters: "7474 x g, 11\xB0C"
  - equipment_name: Vortex Mixer
    settings_parameters: "7843 x g, 22\xB0C"
  procedure_steps:
  - step_description: Cells were transfected with formaldehyde solution to facilitate
      as.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    replicates: 3
  - step_description: Cells were washed with ripa buffer to facilitate again.
    conditions_or_variables:
    - rocking agitation
    - with protease inhibitors
    data_collected: true
    duration_minutes: 691
    temperature_celsius: 26
    replicates: 5
  - step_description: Cells were resolved with formaldehyde solution to facilitate
      himself.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    temperature_celsius: 34
    replicates: 4
control_groups:
- control_type: Negative Control
  description: Stand senior some region choose listen eye lawyer firm one policy artist
    feeling.
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
- Mass spectrometry data processed with MaxQuant
- Flow cytometry data analysis using FlowJo
