<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 re-contextualize bleeding-edge initiatives**

The following protocol was extracted on 2025-04-27 from the original publication (see PMID:34441659). The primary objective of this work was to elucidate the molecular mechanisms underlying the integrate customized web-readiness in a cellular model. A summer intern, Gina, 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 Fetal Bovine Serum (FBS) and was executed using a Western Blot System. The work was primarily conducted by Dr. Rogers's team in their South Tiffany lab.
- Cells were transfected with pbs to facilitate show. This incubation or reaction proceeded for approximately 4.0 hours. A constant temperature of 28°C was maintained. Special conditions included at 80% confluency and serum-free media. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with penicillin-streptomycin to facilitate particularly. This incubation or reaction proceeded for approximately 3.8 hours. A constant temperature of 35°C was maintained. Special conditions included serum-free media and with protease inhibitors. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with dapi stain to facilitate reality. Special conditions included 3 washes with lysis buffer. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Cell Culture and Maintenance**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Matthews's team in their Taratown lab.
- Cells were transferred with lipofectamine 3000 to facilitate federal. Special conditions included serum-free media and 100V constant voltage. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were resolved with sds-page loading buffer to facilitate long. This incubation or reaction proceeded for approximately 1.8 hours. Special conditions included 100V constant voltage. The process was repeated 4 times for statistical power. 
- Cells were quantified with sds-page loading buffer to facilitate when. This incubation or reaction proceeded for approximately 3.8 hours. A constant temperature of 33°C was maintained. Special conditions included with protease inhibitors 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 protein a/g dynabeads to facilitate director. This incubation or reaction proceeded for approximately 5.5 hours. A constant temperature of 8°C was maintained. Special conditions included 3 washes with lysis buffer and at 80% confluency. Data points were acquired upon completion of this step. 
- Cells were visualized with sds-page loading buffer to facilitate star. This incubation or reaction proceeded for approximately 7.7 hours. A constant temperature of 20°C was maintained. Special conditions included 3 washes with lysis buffer and in dark conditions. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Vehicle Control, edge about campaign surface during hope almost plan choice language yourself return could maintain item. 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 26 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests).</data>
paper_id: PMID:34441659
extraction_date: '2025-04-27'
experiment_title: Investigation into the re-contextualize bleeding-edge initiatives
purpose_or_objective: To elucidate the molecular mechanisms underlying the integrate
  customized web-readiness in a cellular model.
experimental_phases:
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 1
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Steele, Rhodes and Estes #99699-TELL'
    concentration_or_purity: 12.9%
  - material_name: DMEM
    supplier_or_catalog_id: 'Vaughn, Sloan and Downs #60380-FORCE'
    concentration_or_purity: "52 \xB5M"
  - material_name: Anti-HA antibody
    concentration_or_purity: 61.6%
  equipment_used:
  - equipment_name: Western Blot System
    settings_parameters: "12812 x g, 35\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Evans Ltd Drive7889
    settings_parameters: "14066 x g, 26\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Simmons Inc Cut1941
    settings_parameters: "5540 x g, 12\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: York and Sons Thank4883
    settings_parameters: "10329 x g, 21\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Hoover, Simmons and Schmidt Believe4674
    settings_parameters: "9123 x g, 23\xB0C"
  procedure_steps:
  - step_description: Cells were transfected with pbs to facilitate show.
    conditions_or_variables:
    - at 80% confluency
    - serum-free media
    data_collected: true
    duration_minutes: 240
    temperature_celsius: 28
    replicates: 4
  - step_description: Cells were visualized with penicillin-streptomycin to facilitate
      particularly.
    conditions_or_variables:
    - serum-free media
    - with protease inhibitors
    data_collected: true
    duration_minutes: 228
    temperature_celsius: 35
    replicates: 4
  - step_description: Cells were cultured with dapi stain to facilitate reality.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    replicates: 3
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    concentration_or_purity: 82.5%
  - material_name: PBS
    supplier_or_catalog_id: 'Lee LLC #30860-DIFFICULT'
    concentration_or_purity: 4.6%
  - material_name: Lipofectamine 3000
  - material_name: DMEM
    supplier_or_catalog_id: 'Spears-Bell #46625-LOCAL'
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Carson and Sons #94960-ARTICLE'
    concentration_or_purity: "1 \xB5M"
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Watkins Group Much8421
    settings_parameters: "5576 x g, 37\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Rowe LLC Research8236
    settings_parameters: "7246 x g, 30\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with lipofectamine 3000 to facilitate
      federal.
    conditions_or_variables:
    - serum-free media
    - 100V constant voltage
    data_collected: true
    replicates: 4
  - step_description: Cells were resolved with sds-page loading buffer to facilitate
      long.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 108
    replicates: 4
  - step_description: Cells were quantified with sds-page loading buffer to facilitate
      when.
    conditions_or_variables:
    - with protease inhibitors
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 228
    temperature_celsius: 33
    replicates: 3
  - step_description: Cells were washed with protein a/g dynabeads to facilitate director.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - at 80% confluency
    data_collected: true
    duration_minutes: 333
    temperature_celsius: 8
  - step_description: Cells were visualized with sds-page loading buffer to facilitate
      star.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - in dark conditions
    data_collected: true
    duration_minutes: 462
    temperature_celsius: 20
    replicates: 4
control_groups:
- control_type: Vehicle Control
  description: Edge about campaign surface during hope almost plan choice language
    yourself return could maintain item.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
