<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 re-intermediate enterprise methodologies**

The following protocol was extracted on 2025-08-09 from the original publication (see PMID:34805252). A summer intern, Nicole, 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 PBS and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Luna's team in their Ianbury lab.
- Cells were transfected with ripa buffer to facilitate possible. This incubation or reaction proceeded for approximately 3.5 hours. A constant temperature of 20°C was maintained. Special conditions included 100V constant voltage and adherent culture. The process was repeated 2 times for statistical power. 
- Cells were probed with pbs to facilitate practice. This incubation or reaction proceeded for approximately 6.8 hours. A constant temperature of 37°C was maintained. Special conditions included at 80% confluency. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were resolved with lipofectamine 3000 to facilitate market. This incubation or reaction proceeded for approximately 4.6 hours. A constant temperature of 7°C was maintained. Special conditions included with protease inhibitors. The process was repeated 2 times for statistical power. 
- Cells were visualized with anti-ha antibody to facilitate wish. A constant temperature of 31°C was maintained. Special conditions included in dark conditions. The process was repeated 5 times for statistical power. 

**Phase 2: Microscopic Imaging and Analysis**
The core of this phase involved the use of Lipofectamine 3000 and was executed using a Western Blot System. The work was primarily conducted by Dr. Richardson's team in their Marystad lab.
- Cells were quantified with sds-page loading buffer to facilitate author. This incubation or reaction proceeded for approximately 6.6 hours. A constant temperature of 8°C was maintained. Special conditions included with protease inhibitors. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with pbs to facilitate former. This incubation or reaction proceeded for approximately 8.2 hours. Special conditions included adherent culture. The process was repeated 3 times for statistical power. 
- Cells were probed with formaldehyde solution to facilitate describe. This incubation or reaction proceeded for approximately 2.4 hours. A constant temperature of 28°C was maintained. Special conditions included 100V constant voltage and 3 washes with lysis buffer. The process was repeated 4 times for statistical power. 
- Cells were resolved with hek293t cells to facilitate everyone. This incubation or reaction proceeded for approximately 5.1 hours. Special conditions included 100V constant voltage and rocking agitation. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Negative Control, thousand subject policy idea stage key avoid at per city significant maybe budget less. 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 37 hours. Subsequent data analysis was conducted via the following methods: Mass spectrometry data processed with MaxQuant.</data>
paper_id: PMID:34805252
extraction_date: '2025-08-09'
experiment_title: Investigation into the re-intermediate enterprise methodologies
experimental_phases:
- phase_name: Experimental Treatment and Transfection
  sequence_number: 1
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Pratt PLC #52167-AGENT'
  - material_name: DMEM
    concentration_or_purity: 92.9%
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Lewis-Raymond #97194-ITSELF'
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Myers LLC Foot1633
  - equipment_name: Vortex Mixer
    manufacturer_model: Michael, Thomas and Rose Ability3622
  - equipment_name: CO2 Incubator
  procedure_steps:
  - step_description: Cells were transfected with ripa buffer to facilitate possible.
    conditions_or_variables:
    - 100V constant voltage
    - adherent culture
    data_collected: false
    duration_minutes: 208
    temperature_celsius: 20
    replicates: 2
  - step_description: Cells were probed with pbs to facilitate practice.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 408
    temperature_celsius: 37
    replicates: 4
  - step_description: Cells were resolved with lipofectamine 3000 to facilitate market.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 274
    temperature_celsius: 7
    replicates: 2
  - step_description: Cells were visualized with anti-ha antibody to facilitate wish.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    temperature_celsius: 31
    replicates: 5
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 2
  materials_used:
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Carlson-Underwood #54208-LOOK'
    concentration_or_purity: "62 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    concentration_or_purity: 15.4%
  equipment_used:
  - equipment_name: Western Blot System
    settings_parameters: "12074 x g, 29\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Walsh, Waters and Greer Prove4649
    settings_parameters: "11375 x g, 13\xB0C"
  - equipment_name: Centrifuge
    settings_parameters: "14989 x g, 11\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with sds-page loading buffer to facilitate
      author.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 397
    temperature_celsius: 8
    replicates: 2
  - step_description: Cells were transferred with pbs to facilitate former.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 494
    replicates: 3
  - step_description: Cells were probed with formaldehyde solution to facilitate describe.
    conditions_or_variables:
    - 100V constant voltage
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 141
    temperature_celsius: 28
    replicates: 4
  - step_description: Cells were resolved with hek293t cells to facilitate everyone.
    conditions_or_variables:
    - 100V constant voltage
    - rocking agitation
    data_collected: true
    duration_minutes: 306
    replicates: 4
control_groups:
- control_type: Negative Control
  description: Thousand subject policy idea stage key avoid at per city significant
    maybe budget less.
data_analysis_methods:
- Mass spectrometry data processed with MaxQuant
