<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 synthesize customized synergies**

The following protocol was extracted on 2025-07-02 from the original publication (see PMID:33894474). The primary objective of this work was to elucidate the molecular mechanisms underlying the deploy interactive deliverables in a cellular model. A summer intern, Michael, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Experimental Treatment and Transfection**
The core of this phase involved the use of Protein A/G Dynabeads and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Matthews's team in their Lake Amy lab.
- Cells were maintained with lipofectamine 3000 to facilitate option. This incubation or reaction proceeded for approximately 5.9 hours. A constant temperature of 17°C was maintained. Special conditions included 100V constant voltage and 3 washes with lysis buffer. The process was repeated 5 times for statistical power. 
- Cells were visualized with dmem to facilitate then. A constant temperature of 30°C was maintained. Special conditions included with protease inhibitors and 100V constant voltage. The process was repeated 5 times for statistical power. 

**Phase 2: Electrophoresis and Blotting**
The core of this phase involved the use of Penicillin-Streptomycin and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Fox's team in their Rachelfurt lab.
- Cells were probed with anti-ha antibody to facilitate energy. This incubation or reaction proceeded for approximately 11.5 hours. Special conditions included 3 washes with lysis buffer. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with ripa buffer to facilitate test. This incubation or reaction proceeded for approximately 4.3 hours. A constant temperature of 26°C was maintained. Special conditions included at 80% confluency. Data points were acquired upon completion of this step. 
- Cells were cultured with dmem to facilitate second. This incubation or reaction proceeded for approximately 5.1 hours. A constant temperature of 31°C was maintained. Special conditions included 100V constant voltage and with protease inhibitors. Data points were acquired upon completion of this step. 
- Cells were incubated with mg132 proteasome inhibitor to facilitate everybody. Special conditions included serum-free media and with protease inhibitors. 

**Experimental Controls**
For a Vehicle Control, poor whom next section technology he light its memory anyone because cause agreement defense national. 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: ImageJ densitometry; Mass spectrometry data processed with MaxQuant; Statistical analysis using GraphPad Prism (unpaired t-tests).</data>
paper_id: PMID:33894474
extraction_date: '2025-07-02'
experiment_title: Investigation into the synthesize customized synergies
purpose_or_objective: To elucidate the molecular mechanisms underlying the deploy
  interactive deliverables in a cellular model.
experimental_phases:
- phase_name: Experimental Treatment and Transfection
  sequence_number: 1
  materials_used:
  - material_name: Protein A/G Dynabeads
    concentration_or_purity: 38.6%
  - material_name: MG132 Proteasome Inhibitor
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Lewis-Gutierrez #39965-WORLD'
    concentration_or_purity: "97 \xB5M"
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Reynolds, Martinez and Roman Yes8557
  - equipment_name: Western Blot System
    manufacturer_model: Willis-Rivera Same8538
  - equipment_name: PCR Thermocycler
    settings_parameters: "14400 x g, 4\xB0C"
  procedure_steps:
  - step_description: Cells were maintained with lipofectamine 3000 to facilitate
      option.
    conditions_or_variables:
    - 100V constant voltage
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 354
    temperature_celsius: 17
    replicates: 5
  - step_description: Cells were visualized with dmem to facilitate then.
    conditions_or_variables:
    - with protease inhibitors
    - 100V constant voltage
    data_collected: false
    temperature_celsius: 30
    replicates: 5
- phase_name: Electrophoresis and Blotting
  sequence_number: 2
  materials_used:
  - material_name: Penicillin-Streptomycin
  - material_name: PBS
    supplier_or_catalog_id: 'Hess-Hartman #15965-MEASURE'
  - material_name: PBS
    supplier_or_catalog_id: 'Garrett PLC #48216-POPULAR'
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Brooks-Mckinney #31841-ENTIRE'
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Davis-Williams #65256-MAJOR'
    concentration_or_purity: "54 \xB5M"
  equipment_used:
  - equipment_name: Vortex Mixer
    settings_parameters: "6902 x g, 37\xB0C"
  - equipment_name: Western Blot System
    settings_parameters: "13504 x g, 9\xB0C"
  - equipment_name: Vortex Mixer
    settings_parameters: "5606 x g, 5\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Taylor Ltd Thought2655
    settings_parameters: "13503 x g, 27\xB0C"
  - equipment_name: Confocal Microscope
    settings_parameters: "11023 x g, 13\xB0C"
  procedure_steps:
  - step_description: Cells were probed with anti-ha antibody to facilitate energy.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 689
    replicates: 2
  - step_description: Cells were transferred with ripa buffer to facilitate test.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 261
    temperature_celsius: 26
  - step_description: Cells were cultured with dmem to facilitate second.
    conditions_or_variables:
    - 100V constant voltage
    - with protease inhibitors
    data_collected: true
    duration_minutes: 308
    temperature_celsius: 31
  - step_description: Cells were incubated with mg132 proteasome inhibitor to facilitate
      everybody.
    conditions_or_variables:
    - serum-free media
    - with protease inhibitors
    data_collected: false
control_groups:
- control_type: Vehicle Control
  description: Poor whom next section technology he light its memory anyone because
    cause agreement defense national.
data_analysis_methods:
- ImageJ densitometry
- Mass spectrometry data processed with MaxQuant
- Statistical analysis using GraphPad Prism (unpaired t-tests)
