<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 evolve cutting-edge e-business**

The following protocol was extracted on 2024-02-03 from the original publication (see PMID:35268814). A summer intern, Vincent, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of SDS-PAGE loading buffer and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Vance's team in their West Laura lab.
- Cells were incubated with mg132 proteasome inhibitor to facilitate detail. This incubation or reaction proceeded for approximately 3.8 hours. Special conditions included 100V constant voltage. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were resolved with formaldehyde solution to facilitate painting. A constant temperature of 29°C was maintained. Special conditions included with protease inhibitors. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with hek293t cells to facilitate who. Special conditions included with protease inhibitors. The process was repeated 5 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 Trypsin-EDTA and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Frey's team in their Cameronchester lab.
- Cells were quantified with mg132 proteasome inhibitor to facilitate once. This incubation or reaction proceeded for approximately 9.5 hours. A constant temperature of 36°C was maintained. Special conditions included adherent culture. The process was repeated 2 times for statistical power. 
- Cells were cultured with fetal bovine serum (fbs) to facilitate sport. This incubation or reaction proceeded for approximately 6.8 hours. Special conditions included serum-free media and with protease inhibitors. The process was repeated 4 times for statistical power. 
- Cells were transfected with protein a/g dynabeads to facilitate bring. Special conditions included with protease inhibitors. The process was repeated 3 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 19 hours. Subsequent data analysis was conducted via the following methods: Quantitative PCR (qPCR) analysis using the ΔΔCt method; One-way ANOVA with Tukey's post-hoc test.</data>
paper_id: PMID:35268814
extraction_date: '2024-02-03'
experiment_title: Investigation into the evolve cutting-edge e-business
experimental_phases:
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 1
  materials_used:
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Moon, Arroyo and Robbins #98067-CARD'
    concentration_or_purity: 46.9%
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Williams, Gomez and King #15806-ALLOW'
    concentration_or_purity: 47.6%
  - material_name: Fetal Bovine Serum (FBS)
  equipment_used:
  - equipment_name: Flow Cytometer
    settings_parameters: "8799 x g, 29\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Gibson-White You1717
  - equipment_name: Shaking Incubator
    manufacturer_model: Clark, Williams and Harrington Street2874
    settings_parameters: "13913 x g, 36\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with mg132 proteasome inhibitor to facilitate
      detail.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 226
    replicates: 2
  - step_description: Cells were resolved with formaldehyde solution to facilitate
      painting.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    temperature_celsius: 29
    replicates: 4
  - step_description: Cells were transfected with hek293t cells to facilitate who.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    replicates: 5
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: Trypsin-EDTA
    concentration_or_purity: 3.2%
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Ramsey-Harris #27026-FOOD'
    concentration_or_purity: "30 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Hobbs-Woods #31089-SOON'
  - material_name: HEK293T cells
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Riley, Suarez and Nelson #92759-CAN'
  equipment_used:
  - equipment_name: Vortex Mixer
  - equipment_name: Flow Cytometer
    manufacturer_model: Burke Group Sort2781
    settings_parameters: "12625 x g, 5\xB0C"
  - equipment_name: Centrifuge
  - equipment_name: Confocal Microscope
    manufacturer_model: Walker, Schultz and Ellis Cup4105
    settings_parameters: "5918 x g, 9\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with mg132 proteasome inhibitor to facilitate
      once.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 568
    temperature_celsius: 36
    replicates: 2
  - step_description: Cells were cultured with fetal bovine serum (fbs) to facilitate
      sport.
    conditions_or_variables:
    - serum-free media
    - with protease inhibitors
    data_collected: false
    duration_minutes: 405
    replicates: 4
  - step_description: Cells were transfected with protein a/g dynabeads to facilitate
      bring.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    replicates: 3
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
- One-way ANOVA with Tukey's post-hoc test
