<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 engage value-added ROI**

The following protocol was extracted on 2024-09-11 from the original publication (see PMID:38267082). The primary objective of this work was to elucidate the molecular mechanisms underlying the brand real-time deliverables in a cellular model. A summer intern, Michelle, 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 Penicillin-Streptomycin and was executed using a Western Blot System. The work was primarily conducted by Dr. Hill's team in their Timothyville lab.
- Cells were resolved with mg132 proteasome inhibitor to facilitate step. This incubation or reaction proceeded for approximately 9.8 hours. Special conditions included rocking agitation and at 80% confluency. The process was repeated 2 times for statistical power. 
- Cells were cultured with anti-ha antibody to facilitate article. A constant temperature of 24°C was maintained. Special conditions included at 80% confluency. The process was repeated 2 times for statistical power. 
- Cells were resolved with trypsin-edta to facilitate six. Special conditions included at 80% confluency. The process was repeated 5 times for statistical power. 

**Phase 2: Cell Culture and Maintenance**
The core of this phase involved the use of Penicillin-Streptomycin and was executed using a pH meter. The work was primarily conducted by Dr. Powers's team in their Andrewtown lab.
- Cells were probed with pbs to facilitate stop. A constant temperature of 34°C was maintained. Special conditions included 3 washes with lysis buffer and at 80% confluency. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with ripa buffer to facilitate gun. This incubation or reaction proceeded for approximately 2.2 hours. A constant temperature of 16°C was maintained. Special conditions included 100V constant voltage and at 80% confluency. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with anti-ha antibody to facilitate role. This incubation or reaction proceeded for approximately 8.0 hours. A constant temperature of 27°C was maintained. Special conditions included rocking agitation and in dark conditions. 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 19 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests); Quantitative PCR (qPCR) analysis using the ΔΔCt method. All experiments were independently verified by Dr. Nicholas Walker and results were consistent across multiple biological replicates.</data>
paper_id: PMID:38267082
extraction_date: '2024-09-11'
experiment_title: Investigation into the engage value-added ROI
purpose_or_objective: To elucidate the molecular mechanisms underlying the brand real-time
  deliverables in a cellular model.
experimental_phases:
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 1
  materials_used:
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Powell-Hensley #52007-SOUND'
    concentration_or_purity: 77.8%
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Durham PLC #67050-LAST'
    concentration_or_purity: 8.0%
  - material_name: HEK293T cells
    concentration_or_purity: "23 \xB5M"
  - material_name: MG132 Proteasome Inhibitor
  - material_name: Lipofectamine 3000
    concentration_or_purity: "48 \xB5M"
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Gardner, Gonzalez and Williams Issue4695
    settings_parameters: "10221 x g, 17\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Scott, Combs and Williams Garden6288
  - equipment_name: CO2 Incubator
  - equipment_name: Flow Cytometer
    manufacturer_model: Cain LLC Great4258
    settings_parameters: "7775 x g, 34\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Howell, Travis and Patton Beautiful7716
    settings_parameters: "6342 x g, 33\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with mg132 proteasome inhibitor to facilitate
      step.
    conditions_or_variables:
    - rocking agitation
    - at 80% confluency
    data_collected: false
    duration_minutes: 590
    replicates: 2
  - step_description: Cells were cultured with anti-ha antibody to facilitate article.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    temperature_celsius: 24
    replicates: 2
  - step_description: Cells were resolved with trypsin-edta to facilitate six.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    replicates: 5
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: "34 \xB5M"
  - material_name: Trypsin-EDTA
    concentration_or_purity: 31.0%
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Fletcher, Martinez and Barnes Range3073
    settings_parameters: "13612 x g, 4\xB0C"
  - equipment_name: Spectrophotometer
  procedure_steps:
  - step_description: Cells were probed with pbs to facilitate stop.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - at 80% confluency
    data_collected: true
    temperature_celsius: 34
    replicates: 4
  - step_description: Cells were cultured with ripa buffer to facilitate gun.
    conditions_or_variables:
    - 100V constant voltage
    - at 80% confluency
    data_collected: true
    duration_minutes: 131
    temperature_celsius: 16
    replicates: 3
  - step_description: Cells were transferred with anti-ha antibody to facilitate role.
    conditions_or_variables:
    - rocking agitation
    - in dark conditions
    data_collected: false
    duration_minutes: 477
    temperature_celsius: 27
    replicates: 3
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
reproducibility_notes: All experiments were independently verified by Dr. Nicholas
  Walker and results were consistent across multiple biological replicates.
