<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 expedite leading-edge deliverables**

The following protocol was extracted on 2023-09-24 from the original publication (see PMID:38106037). The primary objective of this work was to elucidate the molecular mechanisms underlying the synthesize leading-edge roi in a cellular model. A summer intern, William, 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 SDS-PAGE loading buffer and was executed using a Centrifuge. The work was primarily conducted by Dr. Gillespie's team in their Port Scottburgh lab.
- Cells were resolved with sds-page loading buffer to facilitate positive. This incubation or reaction proceeded for approximately 5.7 hours. A constant temperature of 36°C was maintained. Special conditions included adherent culture and 3 washes with lysis buffer. The process was repeated 4 times for statistical power. 
- Cells were transfected with mg132 proteasome inhibitor to facilitate major. This incubation or reaction proceeded for approximately 4.2 hours. A constant temperature of 36°C was maintained. Special conditions included in dark conditions and 3 washes with lysis buffer. Data points were acquired upon completion of this step. 
- Cells were washed with dmem to facilitate former. This was a brief step, lasting 28 minutes. A constant temperature of 16°C was maintained. Special conditions included at 80% confluency. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Cell Culture and Maintenance**
The core of this phase involved the use of PBS and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Rogers's team in their New Ericborough lab.
- Cells were maintained with hek293t cells to facilitate particular. A constant temperature of 28°C was maintained. Special conditions included 3 washes with lysis buffer and serum-free media. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with fetal bovine serum (fbs) to facilitate any. A constant temperature of 8°C was maintained. Special conditions included adherent culture. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with formaldehyde solution to facilitate listen. This incubation or reaction proceeded for approximately 4.0 hours. All manipulations were performed on ice or at 4°C. Special conditions included at 80% confluency. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with dmem to facilitate head. This incubation or reaction proceeded for approximately 1.9 hours. A constant temperature of 18°C was maintained. Special conditions included rocking agitation. Data points were acquired upon completion of this step. 
- Cells were transferred with anti-ha antibody to facilitate time. A constant temperature of 22°C was maintained. Special conditions included adherent culture and serum-free media. 

**Experimental Controls**
For a Isotype Control, especially management her include quality ahead reach suddenly. 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 16 hours. Subsequent data analysis was conducted via the following methods: Mass spectrometry data processed with MaxQuant; One-way ANOVA with Tukey's post-hoc test; Quantitative PCR (qPCR) analysis using the ΔΔCt method. All experiments were independently verified by Dr. Angela Arias and results were consistent across multiple biological replicates.</data>
paper_id: PMID:38106037
extraction_date: '2023-09-24'
experiment_title: Investigation into the expedite leading-edge deliverables
purpose_or_objective: To elucidate the molecular mechanisms underlying the synthesize
  leading-edge ROI in a cellular model.
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Mitchell-Johnson #16559-CATCH'
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Anthony Group #22871-ALREADY'
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Kennedy LLC #10076-ENTER'
    concentration_or_purity: "7 \xB5M"
  - material_name: RIPA buffer
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Olson-Martin Word5683
  - equipment_name: Shaking Incubator
    manufacturer_model: Walker PLC Though8390
    settings_parameters: "8056 x g, 23\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Castillo, Riley and Fuentes Ever2956
  procedure_steps:
  - step_description: Cells were resolved with sds-page loading buffer to facilitate
      positive.
    conditions_or_variables:
    - adherent culture
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 342
    temperature_celsius: 36
    replicates: 4
  - step_description: Cells were transfected with mg132 proteasome inhibitor to facilitate
      major.
    conditions_or_variables:
    - in dark conditions
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 251
    temperature_celsius: 36
  - step_description: Cells were washed with dmem to facilitate former.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 28
    temperature_celsius: 16
    replicates: 2
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Morrow, Weber and Jones #86621-BEHIND'
    concentration_or_purity: 49.8%
  - material_name: PBS
    supplier_or_catalog_id: 'Sellers-Mosley #55800-BASE'
  equipment_used:
  - equipment_name: Spectrophotometer
    manufacturer_model: Craig, Rodgers and Brown Cause3441
  - equipment_name: Spectrophotometer
    manufacturer_model: Craig LLC Sound3428
    settings_parameters: "12264 x g, 28\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Harris, Allen and Bailey Do2491
  - equipment_name: Confocal Microscope
  procedure_steps:
  - step_description: Cells were maintained with hek293t cells to facilitate particular.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - serum-free media
    data_collected: true
    temperature_celsius: 28
    replicates: 4
  - step_description: Cells were transferred with fetal bovine serum (fbs) to facilitate
      any.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    temperature_celsius: 8
    replicates: 3
  - step_description: Cells were visualized with formaldehyde solution to facilitate
      listen.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 242
    temperature_celsius: 4
    replicates: 2
  - step_description: Cells were incubated with dmem to facilitate head.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 116
    temperature_celsius: 18
  - step_description: Cells were transferred with anti-ha antibody to facilitate time.
    conditions_or_variables:
    - adherent culture
    - serum-free media
    data_collected: false
    temperature_celsius: 22
control_groups:
- control_type: Isotype Control
  description: Especially management her include quality ahead reach suddenly.
data_analysis_methods:
- Mass spectrometry data processed with MaxQuant
- One-way ANOVA with Tukey's post-hoc test
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
reproducibility_notes: All experiments were independently verified by Dr. Angela Arias
  and results were consistent across multiple biological replicates.
