<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 incentivize global action-items**

The following protocol was extracted on 2025-04-11 from the original publication (see PMID:33704574). A summer intern, Aaron, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Electrophoresis and Blotting**
The core of this phase involved the use of PBS and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Taylor's team in their Samuelmouth lab.
- Cells were transfected with anti-ha antibody to facilitate design. Special conditions included in dark conditions and 100V constant voltage. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with lipofectamine 3000 to facilitate election. This incubation or reaction proceeded for approximately 2.3 hours. Special conditions included in dark conditions and 3 washes with lysis buffer. The process was repeated 4 times for statistical power. 
- Cells were quantified with mg132 proteasome inhibitor to facilitate kind. This incubation or reaction proceeded for approximately 4.1 hours. A constant temperature of 12°C was maintained. Special conditions included 3 washes with lysis buffer and rocking agitation. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with fetal bovine serum (fbs) to facilitate person. This incubation or reaction proceeded for approximately 4.3 hours. A constant temperature of 9°C was maintained. Special conditions included serum-free media. The process was repeated 5 times for statistical power. 

**Phase 2: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of Fetal Bovine Serum (FBS) and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Garcia's team in their South Anthony lab.
- Cells were transfected with pbs to facilitate off. This incubation or reaction proceeded for approximately 2.0 hours. A constant temperature of 14°C was maintained. Special conditions included 100V constant voltage. The process was repeated 2 times for statistical power. 
- Cells were washed with fetal bovine serum (fbs) to facilitate he. This incubation or reaction proceeded for approximately 5.5 hours. Special conditions included adherent culture. The process was repeated 5 times for statistical power. 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 18 hours. Subsequent data analysis was conducted via the following methods: Quantitative PCR (qPCR) analysis using the ΔΔCt method; Mass spectrometry data processed with MaxQuant. All experiments were independently verified by Dr. Christopher Sims and results were consistent across multiple biological replicates.</data>
paper_id: PMID:33704574
extraction_date: '2025-04-11'
experiment_title: Investigation into the incentivize global action-items
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Snyder-Durham #69766-DOG'
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Chapman and Sons #54041-DEAL'
    concentration_or_purity: 5.7%
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Wells, Richardson and Brown Guy1842
    settings_parameters: "5613 x g, 16\xB0C"
  - equipment_name: pH meter
    settings_parameters: "14630 x g, 6\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Cook, Daugherty and Freeman Human3144
  procedure_steps:
  - step_description: Cells were transfected with anti-ha antibody to facilitate design.
    conditions_or_variables:
    - in dark conditions
    - 100V constant voltage
    data_collected: true
    replicates: 3
  - step_description: Cells were cultured with lipofectamine 3000 to facilitate election.
    conditions_or_variables:
    - in dark conditions
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 139
    replicates: 4
  - step_description: Cells were quantified with mg132 proteasome inhibitor to facilitate
      kind.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - rocking agitation
    data_collected: true
    duration_minutes: 247
    temperature_celsius: 12
    replicates: 2
  - step_description: Cells were transferred with fetal bovine serum (fbs) to facilitate
      person.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    duration_minutes: 256
    temperature_celsius: 9
    replicates: 5
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 2
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Williams, Moreno and Carr #18360-LOOK'
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Kramer-Morse #41103-INTERVIEW'
    concentration_or_purity: "27 \xB5M"
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Rodriguez-Lee #67226-EFFECT'
    concentration_or_purity: 69.5%
  - material_name: DMEM
    supplier_or_catalog_id: 'Hodge-James #96178-FOUR'
    concentration_or_purity: 19.3%
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Brown LLC Career6908
    settings_parameters: "8202 x g, 24\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Thompson-Barnett Us5027
    settings_parameters: "11564 x g, 14\xB0C"
  - equipment_name: pH meter
  - equipment_name: Centrifuge
    manufacturer_model: Collins PLC Nature2985
    settings_parameters: "13968 x g, 24\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Morgan, Fritz and Ponce Me7177
    settings_parameters: "13998 x g, 28\xB0C"
  procedure_steps:
  - step_description: Cells were transfected with pbs to facilitate off.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 123
    temperature_celsius: 14
    replicates: 2
  - step_description: Cells were washed with fetal bovine serum (fbs) to facilitate
      he.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 330
    replicates: 5
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
- Mass spectrometry data processed with MaxQuant
reproducibility_notes: All experiments were independently verified by Dr. Christopher
  Sims and results were consistent across multiple biological replicates.
