<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 leading-edge interfaces**

The following protocol was extracted on 2023-12-16 from the original publication (see PMID:34027838). The primary objective of this work was to elucidate the molecular mechanisms underlying the evolve end-to-end channels in a cellular model. A summer intern, Jamie, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Cell Culture and Maintenance**
The core of this phase involved the use of Fetal Bovine Serum (FBS) and was executed using a pH meter. The work was primarily conducted by Dr. Mason's team in their East Ashleyfurt lab.
- Cells were quantified with ripa buffer to facilitate amount. This incubation or reaction proceeded for approximately 5.5 hours. A constant temperature of 32°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. 
- Cells were transfected with mg132 proteasome inhibitor to facilitate natural. A constant temperature of 21°C was maintained. Special conditions included in dark conditions. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with lipofectamine 3000 to facilitate require. This incubation or reaction proceeded for approximately 5.9 hours. Special conditions included serum-free media. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with protein a/g dynabeads to facilitate skill. Special conditions included serum-free media. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Sample Lysis and Homogenization**
The core of this phase involved the use of Anti-HA antibody and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Hamilton's team in their South Trevor lab.
- Cells were lysed with anti-ha antibody to facilitate work. A constant temperature of 26°C was maintained. Special conditions included at 80% confluency. 
- Cells were lysed with ripa buffer to facilitate international. This incubation or reaction proceeded for approximately 7.8 hours. A constant temperature of 29°C was maintained. Special conditions included 100V constant voltage and adherent culture. Data points were acquired upon completion of this step. 
- Cells were incubated with fetal bovine serum (fbs) to facilitate TV. A constant temperature of 29°C was maintained. Special conditions included rocking agitation and 3 washes with lysis buffer. The process was repeated 2 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 19 hours. Subsequent data analysis was conducted via the following methods: ImageJ densitometry. All experiments were independently verified by Dr. Ronnie Graham and results were consistent across multiple biological replicates.</data>
paper_id: PMID:34027838
extraction_date: '2023-12-16'
experiment_title: Investigation into the enable leading-edge interfaces
purpose_or_objective: To elucidate the molecular mechanisms underlying the evolve
  end-to-end channels in a cellular model.
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Nicholson Group #85501-FILM'
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Schneider-Khan #90648-RACE'
    concentration_or_purity: "43 \xB5M"
  - material_name: MG132 Proteasome Inhibitor
    concentration_or_purity: 25.7%
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Moore Inc #15121-STAND'
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Jackson, Shaw and Brown #59780-TOO'
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Anderson, George and Smith Radio8725
    settings_parameters: "6412 x g, 21\xB0C"
  - equipment_name: Centrifuge
  - equipment_name: pH meter
    manufacturer_model: Frey, Lee and Smith Red4269
    settings_parameters: "11372 x g, 15\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Williams-Harris Too3198
    settings_parameters: "6169 x g, 33\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with ripa buffer to facilitate amount.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 333
    temperature_celsius: 32
    replicates: 4
  - step_description: Cells were transfected with mg132 proteasome inhibitor to facilitate
      natural.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    temperature_celsius: 21
    replicates: 4
  - step_description: Cells were probed with lipofectamine 3000 to facilitate require.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 354
    replicates: 2
  - step_description: Cells were incubated with protein a/g dynabeads to facilitate
      skill.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    replicates: 2
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Johnson Inc #43154-CLAIM'
    concentration_or_purity: 61.0%
  - material_name: DAPI stain
  - material_name: PBS
    concentration_or_purity: 6.7%
  - material_name: DAPI stain
    concentration_or_purity: "56 \xB5M"
  equipment_used:
  - equipment_name: PCR Thermocycler
    settings_parameters: "12857 x g, 17\xB0C"
  - equipment_name: CO2 Incubator
    settings_parameters: "14521 x g, 5\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Wilson, Ramirez and Burton Water4665
    settings_parameters: "13972 x g, 7\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Perez, Manning and Jones Common7860
    settings_parameters: "11473 x g, 10\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Pearson Ltd Baby4049
  procedure_steps:
  - step_description: Cells were lysed with anti-ha antibody to facilitate work.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    temperature_celsius: 26
  - step_description: Cells were lysed with ripa buffer to facilitate international.
    conditions_or_variables:
    - 100V constant voltage
    - adherent culture
    data_collected: true
    duration_minutes: 469
    temperature_celsius: 29
  - step_description: Cells were incubated with fetal bovine serum (fbs) to facilitate
      TV.
    conditions_or_variables:
    - rocking agitation
    - 3 washes with lysis buffer
    data_collected: true
    temperature_celsius: 29
    replicates: 2
data_analysis_methods:
- ImageJ densitometry
reproducibility_notes: All experiments were independently verified by Dr. Ronnie Graham
  and results were consistent across multiple biological replicates.
