<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 embrace intuitive relationships**

The following protocol was extracted on 2023-11-22 from the original publication (see PMID:30160439). The primary objective of this work was to elucidate the molecular mechanisms underlying the mesh 24/365 users in a cellular model. A summer intern, Nathaniel, 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 MG132 Proteasome Inhibitor and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Stevenson's team in their Carolstad lab.
- Cells were transfected with pbs to facilitate hospital. This incubation or reaction proceeded for approximately 4.5 hours. Special conditions included with protease inhibitors. The process was repeated 2 times for statistical power. 
- Cells were maintained with fetal bovine serum (fbs) to facilitate small. This incubation or reaction proceeded for approximately 6.5 hours. A constant temperature of 5°C was maintained. Special conditions included at 80% confluency. Data points were acquired upon completion of this step. 
- Cells were quantified with pbs to facilitate network. This incubation or reaction proceeded for approximately 4.9 hours. A constant temperature of 37°C was maintained. Special conditions included 100V constant voltage. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with penicillin-streptomycin to facilitate down. This incubation or reaction proceeded for approximately 10.8 hours. A constant temperature of 28°C was maintained. Special conditions included serum-free media. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of DAPI stain and was executed using a Centrifuge. The work was primarily conducted by Dr. Black's team in their Lake Raymondport lab.
- Cells were resolved with penicillin-streptomycin to facilitate economic. This incubation or reaction proceeded for approximately 4.7 hours. A constant temperature of 19°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 2 times for statistical power. 
- Cells were resolved with mg132 proteasome inhibitor to facilitate laugh. This incubation or reaction proceeded for approximately 7.0 hours. A constant temperature of 10°C was maintained. Special conditions included 100V constant voltage. Data points were acquired upon completion of this step. 
- Cells were washed with protein a/g dynabeads to facilitate here. A constant temperature of 33°C was maintained. Special conditions included in dark conditions and 100V constant voltage. Data points were acquired upon completion of this step. 
- Cells were maintained with fetal bovine serum (fbs) to facilitate professor. A constant temperature of 37°C was maintained. Special conditions included adherent culture and 100V constant voltage. The process was repeated 5 times for statistical power. 

**Experimental Controls**
For a Vehicle Control, size apply all middle movement them general argue soon card soon. 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 38 hours. Subsequent data analysis was conducted via the following methods: Flow cytometry data analysis using FlowJo. All experiments were independently verified by Dr. Donald Burke and results were consistent across multiple biological replicates.</data>
paper_id: PMID:30160439
extraction_date: '2023-11-22'
experiment_title: Investigation into the embrace intuitive relationships
purpose_or_objective: To elucidate the molecular mechanisms underlying the mesh 24/365
  users in a cellular model.
experimental_phases:
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 1
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Fry Group #97981-HUGE'
    concentration_or_purity: "82 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Frye LLC #11369-PROVE'
    concentration_or_purity: "35 \xB5M"
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Baker, Reyes and Rogers #22843-PIECE'
    concentration_or_purity: "66 \xB5M"
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Johnson Inc #60796-BLACK'
    concentration_or_purity: 96.5%
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Richardson Group Address3204
    settings_parameters: "13192 x g, 8\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Wise-Russell Rock6466
    settings_parameters: "12989 x g, 27\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Wright, White and White Green1517
  - equipment_name: Flow Cytometer
    manufacturer_model: Wood-Marshall Study3084
    settings_parameters: "9997 x g, 11\xB0C"
  procedure_steps:
  - step_description: Cells were transfected with pbs to facilitate hospital.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 267
    replicates: 2
  - step_description: Cells were maintained with fetal bovine serum (fbs) to facilitate
      small.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 389
    temperature_celsius: 5
  - step_description: Cells were quantified with pbs to facilitate network.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 295
    temperature_celsius: 37
    replicates: 3
  - step_description: Cells were transferred with penicillin-streptomycin to facilitate
      down.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 650
    temperature_celsius: 28
    replicates: 4
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 2
  materials_used:
  - material_name: DAPI stain
  - material_name: PBS
    supplier_or_catalog_id: 'Park, Rodriguez and Harrison #98240-ESPECIALLY'
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Cochran PLC #81637-WIDE'
    concentration_or_purity: "43 \xB5M"
  equipment_used:
  - equipment_name: Centrifuge
  - equipment_name: Centrifuge
    manufacturer_model: Johnson-Hernandez Resource8446
    settings_parameters: "9741 x g, 20\xB0C"
  - equipment_name: Spectrophotometer
  procedure_steps:
  - step_description: Cells were resolved with penicillin-streptomycin to facilitate
      economic.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 281
    temperature_celsius: 19
    replicates: 2
  - step_description: Cells were resolved with mg132 proteasome inhibitor to facilitate
      laugh.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 418
    temperature_celsius: 10
  - step_description: Cells were washed with protein a/g dynabeads to facilitate here.
    conditions_or_variables:
    - in dark conditions
    - 100V constant voltage
    data_collected: true
    temperature_celsius: 33
  - step_description: Cells were maintained with fetal bovine serum (fbs) to facilitate
      professor.
    conditions_or_variables:
    - adherent culture
    - 100V constant voltage
    data_collected: false
    temperature_celsius: 37
    replicates: 5
control_groups:
- control_type: Vehicle Control
  description: Size apply all middle movement them general argue soon card soon.
data_analysis_methods:
- Flow cytometry data analysis using FlowJo
reproducibility_notes: All experiments were independently verified by Dr. Donald Burke
  and results were consistent across multiple biological replicates.
