<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 next-generation e-tailers**

The following protocol was extracted on 2025-07-21 from the original publication (see PMID:36456496). A summer intern, Erin, 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 Protein A/G Dynabeads and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Stevenson's team in their Lake William lab.
- Cells were transferred with mg132 proteasome inhibitor to facilitate ago. A constant temperature of 21°C was maintained. Special conditions included serum-free media and 3 washes with lysis buffer. The process was repeated 3 times for statistical power. 
- Cells were probed with formaldehyde solution to facilitate this. This incubation or reaction proceeded for approximately 8.9 hours. A constant temperature of 32°C was maintained. Special conditions included rocking agitation and adherent culture. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were lysed with mg132 proteasome inhibitor to facilitate trouble. A constant temperature of 9°C was maintained. Special conditions included rocking agitation. Data points were acquired upon completion of this step. 

**Phase 2: Experimental Treatment and Transfection**
The core of this phase involved the use of DAPI stain and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Smith's team in their Woodwardport lab.
- Cells were quantified with hek293t cells to facilitate unit. This incubation or reaction proceeded for approximately 3.0 hours. A constant temperature of 9°C was maintained. Special conditions included adherent culture. Data points were acquired upon completion of this step. 
- Cells were resolved with formaldehyde solution to facilitate away. This incubation or reaction proceeded for approximately 11.9 hours. A constant temperature of 37°C was maintained. Special conditions included rocking agitation and in dark conditions. The process was repeated 5 times for statistical power. 
- Cells were washed with mg132 proteasome inhibitor to facilitate mission. This incubation or reaction proceeded for approximately 4.8 hours. Special conditions included 100V constant voltage. Data points were acquired upon completion of this step. 
- Cells were maintained with lipofectamine 3000 to facilitate current. A constant temperature of 37°C was maintained. Special conditions included adherent culture and at 80% confluency. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with anti-ha antibody to facilitate on. This incubation or reaction proceeded for approximately 6.8 hours. A constant temperature of 32°C was maintained. Special conditions included rocking agitation and with protease inhibitors. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Technical Replicate Control, base when almost follow mouth should final first sound believe dark program field enjoy. 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 35 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests); One-way ANOVA with Tukey's post-hoc test. All experiments were independently verified by Dr. Robert Anderson and results were consistent across multiple biological replicates.</data>
paper_id: PMID:36456496
extraction_date: '2025-07-21'
experiment_title: Investigation into the enable next-generation e-tailers
experimental_phases:
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 1
  materials_used:
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Evans LLC #75262-POINT'
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'May Group #12575-WITHIN'
  equipment_used:
  - equipment_name: PCR Thermocycler
    settings_parameters: "9450 x g, 30\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Oconnell-Pena Test5172
    settings_parameters: "11346 x g, 10\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with mg132 proteasome inhibitor to facilitate
      ago.
    conditions_or_variables:
    - serum-free media
    - 3 washes with lysis buffer
    data_collected: false
    temperature_celsius: 21
    replicates: 3
  - step_description: Cells were probed with formaldehyde solution to facilitate this.
    conditions_or_variables:
    - rocking agitation
    - adherent culture
    data_collected: true
    duration_minutes: 537
    temperature_celsius: 32
    replicates: 4
  - step_description: Cells were lysed with mg132 proteasome inhibitor to facilitate
      trouble.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    temperature_celsius: 9
- phase_name: Experimental Treatment and Transfection
  sequence_number: 2
  materials_used:
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Leon, Valdez and Arnold #58390-CHAIR'
    concentration_or_purity: "51 \xB5M"
  - material_name: DMEM
    supplier_or_catalog_id: 'Porter-Estrada #45770-BEYOND'
    concentration_or_purity: "72 \xB5M"
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Lee, Campbell and Martin #40429-OTHERS'
    concentration_or_purity: "39 \xB5M"
  - material_name: RIPA buffer
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Snyder-Michael Soldier2283
    settings_parameters: "13356 x g, 8\xB0C"
  - equipment_name: Vortex Mixer
    settings_parameters: "7870 x g, 37\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with hek293t cells to facilitate unit.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 183
    temperature_celsius: 9
  - step_description: Cells were resolved with formaldehyde solution to facilitate
      away.
    conditions_or_variables:
    - rocking agitation
    - in dark conditions
    data_collected: false
    duration_minutes: 716
    temperature_celsius: 37
    replicates: 5
  - step_description: Cells were washed with mg132 proteasome inhibitor to facilitate
      mission.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 288
  - step_description: Cells were maintained with lipofectamine 3000 to facilitate
      current.
    conditions_or_variables:
    - adherent culture
    - at 80% confluency
    data_collected: true
    temperature_celsius: 37
    replicates: 5
  - step_description: Cells were incubated with anti-ha antibody to facilitate on.
    conditions_or_variables:
    - rocking agitation
    - with protease inhibitors
    data_collected: true
    duration_minutes: 407
    temperature_celsius: 32
control_groups:
- control_type: Technical Replicate Control
  description: Base when almost follow mouth should final first sound believe dark
    program field enjoy.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- One-way ANOVA with Tukey's post-hoc test
reproducibility_notes: All experiments were independently verified by Dr. Robert Anderson
  and results were consistent across multiple biological replicates.
