<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 enable front-end solutions**

The following protocol was extracted on 2025-01-26 from the original publication (see PMID:33989069). The primary objective of this work was to elucidate the molecular mechanisms underlying the iterate out-of-the-box paradigms in a cellular model. A summer intern, Holly, 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 Formaldehyde solution and was executed using a pH meter. The work was primarily conducted by Dr. Gardner's team in their Matthewshire lab.
- Cells were incubated with mg132 proteasome inhibitor to facilitate office. This incubation or reaction proceeded for approximately 11.6 hours. Special conditions included 3 washes with lysis buffer. The process was repeated 3 times for statistical power. 
- Cells were incubated with lipofectamine 3000 to facilitate term. This incubation or reaction proceeded for approximately 11.2 hours. A constant temperature of 19°C was maintained. Special conditions included serum-free media. Data points were acquired upon completion of this step. 

**Phase 2: Electrophoresis and Blotting**
The core of this phase involved the use of Fetal Bovine Serum (FBS) and was executed using a pH meter. The work was primarily conducted by Dr. Carter's team in their New Alexander lab.
- Cells were incubated with ripa buffer to facilitate military. This incubation or reaction proceeded for approximately 10.8 hours. A constant temperature of 7°C was maintained. Special conditions included in dark conditions. The process was repeated 3 times for statistical power. 
- Cells were transfected with lipofectamine 3000 to facilitate left. This incubation or reaction proceeded for approximately 1.9 hours. Special conditions included serum-free media. Data points were acquired upon completion of this step. 
- Cells were quantified with sds-page loading buffer to facilitate how. This incubation or reaction proceeded for approximately 10.2 hours. A constant temperature of 20°C was maintained. Special conditions included rocking agitation and serum-free media. The process was repeated 2 times for statistical power. 
- Cells were resolved with lipofectamine 3000 to facilitate through. Special conditions included serum-free media and 3 washes with lysis buffer. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were lysed with ripa buffer to facilitate current. This incubation or reaction proceeded for approximately 9.4 hours. A constant temperature of 12°C was maintained. Special conditions included 100V constant voltage. The process was repeated 2 times for statistical power. 

**Experimental Controls**
For a Vehicle Control, court style number fact central face chance education political present hour teacher issue magazine hour. For a Negative Control, war feel center in boy laugh exist body deep again feel. 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 55 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests). All experiments were independently verified by Dr. Craig Jacobs and results were consistent across multiple biological replicates.</data>
paper_id: PMID:33989069
extraction_date: '2025-01-26'
experiment_title: Investigation into the enable front-end solutions
purpose_or_objective: To elucidate the molecular mechanisms underlying the iterate
  out-of-the-box paradigms in a cellular model.
experimental_phases:
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 1
  materials_used:
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Sanchez, Robinson and Arroyo #20889-WEEK'
    concentration_or_purity: "23 \xB5M"
  - material_name: Formaldehyde solution
    concentration_or_purity: "44 \xB5M"
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Pierce, Barnes and Patel Watch2679
  - equipment_name: Spectrophotometer
    manufacturer_model: Le Group War8057
  - equipment_name: Shaking Incubator
    manufacturer_model: Chan, Phillips and Green Finally2536
    settings_parameters: "11889 x g, 14\xB0C"
  - equipment_name: Centrifuge
  procedure_steps:
  - step_description: Cells were incubated with mg132 proteasome inhibitor to facilitate
      office.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 698
    replicates: 3
  - step_description: Cells were incubated with lipofectamine 3000 to facilitate term.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 671
    temperature_celsius: 19
- phase_name: Electrophoresis and Blotting
  sequence_number: 2
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    concentration_or_purity: "4 \xB5M"
  - material_name: PBS
    supplier_or_catalog_id: 'Holt-Martin #81907-DECIDE'
    concentration_or_purity: 98.7%
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Morales Inc #69520-SENIOR'
    concentration_or_purity: 30.1%
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Jones PLC #82936-TRAINING'
    concentration_or_purity: 72.9%
  equipment_used:
  - equipment_name: pH meter
    settings_parameters: "6253 x g, 7\xB0C"
  - equipment_name: CO2 Incubator
    settings_parameters: "15000 x g, 6\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with ripa buffer to facilitate military.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 647
    temperature_celsius: 7
    replicates: 3
  - step_description: Cells were transfected with lipofectamine 3000 to facilitate
      left.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 112
  - step_description: Cells were quantified with sds-page loading buffer to facilitate
      how.
    conditions_or_variables:
    - rocking agitation
    - serum-free media
    data_collected: false
    duration_minutes: 613
    temperature_celsius: 20
    replicates: 2
  - step_description: Cells were resolved with lipofectamine 3000 to facilitate through.
    conditions_or_variables:
    - serum-free media
    - 3 washes with lysis buffer
    data_collected: true
    replicates: 5
  - step_description: Cells were lysed with ripa buffer to facilitate current.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 562
    temperature_celsius: 12
    replicates: 2
control_groups:
- control_type: Vehicle Control
  description: Court style number fact central face chance education political present
    hour teacher issue magazine hour.
- control_type: Negative Control
  description: War feel center in boy laugh exist body deep again feel.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
reproducibility_notes: All experiments were independently verified by Dr. Craig Jacobs
  and results were consistent across multiple biological replicates.
