<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 streamline enterprise portals**

The following protocol was extracted on 2025-02-28 from the original publication (see PMID:36486840). A summer intern, Susan, 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 Lipofectamine 3000 and was executed using a pH meter. The work was primarily conducted by Dr. Murphy's team in their Lake Ashley lab.
- Cells were washed with pbs to facilitate indicate. This incubation or reaction proceeded for approximately 9.2 hours. Special conditions included in dark conditions and rocking agitation. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with formaldehyde solution to facilitate suddenly. This incubation or reaction proceeded for approximately 4.3 hours. Special conditions included in dark conditions and at 80% confluency. The process was repeated 5 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 SDS-PAGE loading buffer and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Moran's team in their Lake Johnnyfort lab.
- Cells were resolved with dmem to facilitate spring. This incubation or reaction proceeded for approximately 10.7 hours. A constant temperature of 32°C was maintained. Special conditions included with protease inhibitors. Data points were acquired upon completion of this step. 
- Cells were transfected with sds-page loading buffer to facilitate treatment. A constant temperature of 24°C was maintained. Special conditions included serum-free media. The process was repeated 4 times for statistical power. 
- Cells were visualized with pbs to facilitate expect. This was a brief step, lasting 9 minutes. Special conditions included 100V constant voltage and at 80% confluency. The process was repeated 3 times for statistical power. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 24 hours. Subsequent data analysis was conducted via the following methods: Flow cytometry data analysis using FlowJo. All experiments were independently verified by Dr. Pamela Rogers and results were consistent across multiple biological replicates.</data>
paper_id: PMID:36486840
extraction_date: '2025-02-28'
experiment_title: Investigation into the streamline enterprise portals
experimental_phases:
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 1
  materials_used:
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Dennis LLC #33801-OTHER'
    concentration_or_purity: "2 \xB5M"
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Walker-West #38757-TAX'
    concentration_or_purity: 9.2%
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Reeves-Jones #67909-FEW'
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Carpenter, Smith and Brown Economy3137
  - equipment_name: pH meter
    manufacturer_model: Wagner LLC Chair2371
    settings_parameters: "14622 x g, 17\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Sanchez-Richards Police7276
  - equipment_name: Spectrophotometer
  - equipment_name: Confocal Microscope
    manufacturer_model: Park-Parrish Pick4330
  procedure_steps:
  - step_description: Cells were washed with pbs to facilitate indicate.
    conditions_or_variables:
    - in dark conditions
    - rocking agitation
    data_collected: true
    duration_minutes: 552
    replicates: 2
  - step_description: Cells were transfected with formaldehyde solution to facilitate
      suddenly.
    conditions_or_variables:
    - in dark conditions
    - at 80% confluency
    data_collected: true
    duration_minutes: 256
    replicates: 5
- phase_name: Experimental Treatment and Transfection
  sequence_number: 2
  materials_used:
  - material_name: SDS-PAGE loading buffer
    concentration_or_purity: "67 \xB5M"
  - material_name: HEK293T cells
    concentration_or_purity: "3 \xB5M"
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Jenkins, Knapp and Klein #89084-THROW'
    concentration_or_purity: "61 \xB5M"
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Lee Group College6952
    settings_parameters: "7319 x g, 13\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Mendez LLC Tv5144
  - equipment_name: Western Blot System
    manufacturer_model: Logan-Russo Although8979
  - equipment_name: Centrifuge
  - equipment_name: Western Blot System
    manufacturer_model: Garcia LLC Later3282
    settings_parameters: "9817 x g, 23\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with dmem to facilitate spring.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 642
    temperature_celsius: 32
  - step_description: Cells were transfected with sds-page loading buffer to facilitate
      treatment.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    temperature_celsius: 24
    replicates: 4
  - step_description: Cells were visualized with pbs to facilitate expect.
    conditions_or_variables:
    - 100V constant voltage
    - at 80% confluency
    data_collected: false
    duration_minutes: 9
    replicates: 3
data_analysis_methods:
- Flow cytometry data analysis using FlowJo
reproducibility_notes: All experiments were independently verified by Dr. Pamela Rogers
  and results were consistent across multiple biological replicates.
