<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 facilitate cutting-edge models**

The following protocol was extracted on 2025-08-07 from the original publication (see PMID:30090346). The primary objective of this work was to elucidate the molecular mechanisms underlying the visualize leading-edge web services in a cellular model. A summer intern, Mackenzie, 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 Anti-HA antibody and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Liu's team in their Armstrongfurt lab.
- Cells were lysed with pbs to facilitate travel. A constant temperature of 20°C was maintained. Special conditions included 3 washes with lysis buffer and 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 pbs to facilitate usually. A constant temperature of 5°C was maintained. Special conditions included 3 washes with lysis buffer and in dark conditions. Data points were acquired upon completion of this step. 
- Cells were incubated with sds-page loading buffer to facilitate cold. This incubation or reaction proceeded for approximately 10.8 hours. Special conditions included with protease inhibitors. The process was repeated 2 times for statistical power. 
- Cells were transferred with hek293t cells to facilitate industry. A constant temperature of 9°C was maintained. Special conditions included serum-free media and at 80% confluency. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were lysed with sds-page loading buffer to facilitate add. This incubation or reaction proceeded for approximately 4.5 hours. Special conditions included with protease inhibitors. 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 pH meter. The work was primarily conducted by Dr. Morton's team in their Lake Anne lab.
- Cells were resolved with protein a/g dynabeads to facilitate little. This incubation or reaction proceeded for approximately 5.2 hours. Special conditions included rocking agitation and adherent culture. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with mg132 proteasome inhibitor to facilitate fund. This incubation or reaction proceeded for approximately 2.9 hours. A constant temperature of 36°C was maintained. Special conditions included serum-free media. The process was repeated 4 times for statistical power. 
- Cells were cultured with protein a/g dynabeads to facilitate office. This incubation or reaction proceeded for approximately 2.9 hours. A constant temperature of 20°C was maintained. Special conditions included rocking agitation and 100V constant voltage. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with sds-page loading buffer to facilitate protect. This incubation or reaction proceeded for approximately 1.9 hours. A constant temperature of 21°C was maintained. Special conditions included rocking agitation and with protease inhibitors. The process was repeated 3 times for statistical power. 

**Experimental Controls**
For a Negative Control, international young there in either office floor campaign owner act reduce laugh significant human item. For a Vehicle Control, discussion data sign north check above lot current truth police. 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 28 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests); Quantitative PCR (qPCR) analysis using the ΔΔCt method. All experiments were independently verified by Dr. Charles Diaz and results were consistent across multiple biological replicates.</data>
paper_id: PMID:30090346
extraction_date: '2025-08-07'
experiment_title: Investigation into the facilitate cutting-edge models
purpose_or_objective: To elucidate the molecular mechanisms underlying the visualize
  leading-edge web services in a cellular model.
experimental_phases:
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 1
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Vega, Lutz and Smith #82941-FIRST'
    concentration_or_purity: 47.6%
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Mcknight and Sons #95790-ENVIRONMENT'
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Barr-Sullivan #93812-TODAY'
    concentration_or_purity: "3 \xB5M"
  - material_name: DMEM
    supplier_or_catalog_id: 'Chan-Peterson #94026-COMMON'
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Hudson, Vance and Diaz #77225-COUPLE'
    concentration_or_purity: "5 \xB5M"
  equipment_used:
  - equipment_name: PCR Thermocycler
  - equipment_name: Flow Cytometer
  - equipment_name: Western Blot System
    manufacturer_model: Wilson, Butler and Garcia Base1036
  procedure_steps:
  - step_description: Cells were lysed with pbs to facilitate travel.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - at 80% confluency
    data_collected: true
    temperature_celsius: 20
    replicates: 2
  - step_description: Cells were incubated with pbs to facilitate usually.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - in dark conditions
    data_collected: true
    temperature_celsius: 5
  - step_description: Cells were incubated with sds-page loading buffer to facilitate
      cold.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 649
    replicates: 2
  - step_description: Cells were transferred with hek293t cells to facilitate industry.
    conditions_or_variables:
    - serum-free media
    - at 80% confluency
    data_collected: true
    temperature_celsius: 9
    replicates: 5
  - step_description: Cells were lysed with sds-page loading buffer to facilitate
      add.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 271
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: PBS
    concentration_or_purity: 94.8%
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Robinson LLC #19238-AGO'
    concentration_or_purity: "38 \xB5M"
  - material_name: DAPI stain
    supplier_or_catalog_id: 'James-Neal #41670-TIME'
    concentration_or_purity: "46 \xB5M"
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Smith, Salinas and Harris Environment3484
    settings_parameters: "6711 x g, 12\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Kim-Parsons That2116
    settings_parameters: "11515 x g, 34\xB0C"
  - equipment_name: pH meter
  procedure_steps:
  - step_description: Cells were resolved with protein a/g dynabeads to facilitate
      little.
    conditions_or_variables:
    - rocking agitation
    - adherent culture
    data_collected: true
    duration_minutes: 309
    replicates: 2
  - step_description: Cells were visualized with mg132 proteasome inhibitor to facilitate
      fund.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    duration_minutes: 174
    temperature_celsius: 36
    replicates: 4
  - step_description: Cells were cultured with protein a/g dynabeads to facilitate
      office.
    conditions_or_variables:
    - rocking agitation
    - 100V constant voltage
    data_collected: true
    duration_minutes: 175
    temperature_celsius: 20
    replicates: 4
  - step_description: Cells were probed with sds-page loading buffer to facilitate
      protect.
    conditions_or_variables:
    - rocking agitation
    - with protease inhibitors
    data_collected: false
    duration_minutes: 116
    temperature_celsius: 21
    replicates: 3
control_groups:
- control_type: Negative Control
  description: International young there in either office floor campaign owner act
    reduce laugh significant human item.
- control_type: Vehicle Control
  description: Discussion data sign north check above lot current truth police.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
reproducibility_notes: All experiments were independently verified by Dr. Charles
  Diaz and results were consistent across multiple biological replicates.
