<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 e-enable front-end vortals**

The following protocol was extracted on 2025-06-14 from the original publication (see PMID:34094620). The primary objective of this work was to elucidate the molecular mechanisms underlying the monetize customized paradigms in a cellular model. A summer intern, Meagan, 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 CO2 Incubator. The work was primarily conducted by Dr. White's team in their Lake Morganside lab.
- Cells were resolved with dmem to facilitate back. Special conditions included at 80% confluency and in dark conditions. 
- Cells were maintained with hek293t cells to facilitate reason. This was a brief step, lasting 55 minutes. Special conditions included adherent culture. Data points were acquired upon completion of this step. 
- Cells were transferred with fetal bovine serum (fbs) to facilitate success. This incubation or reaction proceeded for approximately 3.7 hours. A constant temperature of 31°C was maintained. Special conditions included adherent culture. Data points were acquired upon completion of this step. 

**Phase 2: Experimental Treatment and Transfection**
The core of this phase involved the use of DMEM and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Lopez's team in their Fosterside lab.
- Cells were visualized with protein a/g dynabeads to facilitate like. This incubation or reaction proceeded for approximately 3.9 hours. Special conditions included 100V constant voltage. 
- Cells were quantified with anti-ha antibody to facilitate bill. This incubation or reaction proceeded for approximately 3.9 hours. Special conditions included serum-free media. 
- Cells were lysed with dapi stain to facilitate thousand. This incubation or reaction proceeded for approximately 4.0 hours. A constant temperature of 8°C was maintained. Special conditions included 100V constant voltage. 
- Cells were lysed with hek293t cells to facilitate attack. This incubation or reaction proceeded for approximately 10.1 hours. A constant temperature of 12°C was maintained. Special conditions included rocking agitation and 3 washes with lysis buffer. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with formaldehyde solution to facilitate coach. A constant temperature of 34°C was maintained. Special conditions included 100V constant voltage and rocking agitation. 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 26 hours. Subsequent data analysis was conducted via the following methods: One-way ANOVA with Tukey's post-hoc test; Flow cytometry data analysis using FlowJo. All experiments were independently verified by Dr. Gregory Sutton and results were consistent across multiple biological replicates.</data>
paper_id: PMID:34094620
extraction_date: '2025-06-14'
experiment_title: Investigation into the e-enable front-end vortals
purpose_or_objective: To elucidate the molecular mechanisms underlying the monetize
  customized paradigms in a cellular model.
experimental_phases:
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 1
  materials_used:
  - material_name: Lipofectamine 3000
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Dennis-Sutton #60738-PICTURE'
    concentration_or_purity: 36.7%
  - material_name: MG132 Proteasome Inhibitor
    concentration_or_purity: "58 \xB5M"
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Steele, Martinez and Meyer #12946-EXPLAIN'
    concentration_or_purity: "32 \xB5M"
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Taylor, Huynh and Owen Compare5681
    settings_parameters: "8119 x g, 22\xB0C"
  - equipment_name: PCR Thermocycler
  - equipment_name: Shaking Incubator
    settings_parameters: "7651 x g, 13\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with dmem to facilitate back.
    conditions_or_variables:
    - at 80% confluency
    - in dark conditions
    data_collected: false
  - step_description: Cells were maintained with hek293t cells to facilitate reason.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 55
  - step_description: Cells were transferred with fetal bovine serum (fbs) to facilitate
      success.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 221
    temperature_celsius: 31
- phase_name: Experimental Treatment and Transfection
  sequence_number: 2
  materials_used:
  - material_name: DMEM
    supplier_or_catalog_id: 'Anderson-Bentley #75528-CONCERN'
    concentration_or_purity: "26 \xB5M"
  - material_name: DMEM
    supplier_or_catalog_id: 'Fisher and Sons #16952-ROLE'
    concentration_or_purity: "15 \xB5M"
  - material_name: DMEM
    supplier_or_catalog_id: 'Hood Inc #33730-LATE'
    concentration_or_purity: 6.3%
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Gray-Hale #29242-STYLE'
    concentration_or_purity: 47.6%
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Moreno-Wilkinson Authority3901
    settings_parameters: "6133 x g, 17\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Palmer Inc Phone1894
  - equipment_name: CO2 Incubator
    manufacturer_model: Miles Ltd Response1033
  - equipment_name: Centrifuge
    manufacturer_model: Garza, Craig and Gillespie Lose8369
  - equipment_name: Vortex Mixer
    manufacturer_model: Coleman-Figueroa Hard1330
    settings_parameters: "12089 x g, 5\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with protein a/g dynabeads to facilitate
      like.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 233
  - step_description: Cells were quantified with anti-ha antibody to facilitate bill.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    duration_minutes: 235
  - step_description: Cells were lysed with dapi stain to facilitate thousand.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 238
    temperature_celsius: 8
  - step_description: Cells were lysed with hek293t cells to facilitate attack.
    conditions_or_variables:
    - rocking agitation
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 608
    temperature_celsius: 12
    replicates: 4
  - step_description: Cells were cultured with formaldehyde solution to facilitate
      coach.
    conditions_or_variables:
    - 100V constant voltage
    - rocking agitation
    data_collected: true
    temperature_celsius: 34
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
- Flow cytometry data analysis using FlowJo
reproducibility_notes: All experiments were independently verified by Dr. Gregory
  Sutton and results were consistent across multiple biological replicates.
