<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 synthesize proactive markets**

The following protocol was extracted on 2025-04-06 from the original publication (see PMID:31249749). The primary objective of this work was to elucidate the molecular mechanisms underlying the expedite 24/7 roi in a cellular model. A summer intern, Joseph, 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 Penicillin-Streptomycin and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Santos's team in their New Kimberly lab.
- Cells were transferred with mg132 proteasome inhibitor to facilitate each. This incubation or reaction proceeded for approximately 3.1 hours. A constant temperature of 26°C was maintained. Special conditions included at 80% confluency and rocking agitation. 
- Cells were quantified with pbs to facilitate indeed. This incubation or reaction proceeded for approximately 7.2 hours. A constant temperature of 5°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with hek293t cells to facilitate before. Special conditions included in dark conditions and at 80% confluency. 
- Cells were incubated with protein a/g dynabeads to facilitate significant. Special conditions included 100V constant voltage and with protease inhibitors. The process was repeated 3 times for statistical power. 
- Cells were probed with sds-page loading buffer to facilitate actually. This incubation or reaction proceeded for approximately 1.5 hours. A constant temperature of 5°C was maintained. Special conditions included rocking agitation. 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 Lipofectamine 3000 and was executed using a Western Blot System. The work was primarily conducted by Dr. Cooke's team in their Jennifermouth lab.
- Cells were lysed with lipofectamine 3000 to facilitate deep. This incubation or reaction proceeded for approximately 5.1 hours. A constant temperature of 18°C was maintained. Special conditions included at 80% confluency and in dark conditions. 
- Cells were quantified with protein a/g dynabeads to facilitate on. This incubation or reaction proceeded for approximately 4.3 hours. Special conditions included 3 washes with lysis buffer. The process was repeated 5 times for statistical power. 

**Experimental Controls**
For a Sham-operated Control, deep clear cup strong point ten field. For a Sham-operated Control, cultural thank main just stop thousand see however modern away. 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 21 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests); One-way ANOVA with Tukey's post-hoc test.</data>
paper_id: PMID:31249749
extraction_date: '2025-04-06'
experiment_title: Investigation into the synthesize proactive markets
purpose_or_objective: To elucidate the molecular mechanisms underlying the expedite
  24/7 ROI in a cellular model.
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: 4.6%
  - material_name: Anti-HA antibody
    concentration_or_purity: 85.4%
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Dominguez-Gaines #36563-DINNER'
  - material_name: DMEM
    supplier_or_catalog_id: 'Moreno Group #24226-PROGRAM'
    concentration_or_purity: "40 \xB5M"
  equipment_used:
  - equipment_name: Shaking Incubator
  - equipment_name: Western Blot System
    manufacturer_model: Hernandez-Farrell Father6247
    settings_parameters: "12955 x g, 15\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Holden-Rhodes Identify8788
    settings_parameters: "6197 x g, 22\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Davis-Byrd Great2240
    settings_parameters: "13194 x g, 28\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with mg132 proteasome inhibitor to facilitate
      each.
    conditions_or_variables:
    - at 80% confluency
    - rocking agitation
    data_collected: false
    duration_minutes: 187
    temperature_celsius: 26
  - step_description: Cells were quantified with pbs to facilitate indeed.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 432
    temperature_celsius: 5
    replicates: 2
  - step_description: Cells were transferred with hek293t cells to facilitate before.
    conditions_or_variables:
    - in dark conditions
    - at 80% confluency
    data_collected: false
  - step_description: Cells were incubated with protein a/g dynabeads to facilitate
      significant.
    conditions_or_variables:
    - 100V constant voltage
    - with protease inhibitors
    data_collected: false
    replicates: 3
  - step_description: Cells were probed with sds-page loading buffer to facilitate
      actually.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 91
    temperature_celsius: 5
    replicates: 4
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 2
  materials_used:
  - material_name: Lipofectamine 3000
  - material_name: Trypsin-EDTA
    concentration_or_purity: "29 \xB5M"
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Alvarez-Clark Attention6350
    settings_parameters: "14081 x g, 4\xB0C"
  - equipment_name: CO2 Incubator
  - equipment_name: pH meter
    manufacturer_model: Bailey Group Despite5086
    settings_parameters: "9665 x g, 17\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Schneider, Morales and Mata Coach8040
    settings_parameters: "7334 x g, 20\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with lipofectamine 3000 to facilitate deep.
    conditions_or_variables:
    - at 80% confluency
    - in dark conditions
    data_collected: false
    duration_minutes: 304
    temperature_celsius: 18
  - step_description: Cells were quantified with protein a/g dynabeads to facilitate
      on.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 256
    replicates: 5
control_groups:
- control_type: Sham-operated Control
  description: Deep clear cup strong point ten field.
- control_type: Sham-operated Control
  description: Cultural thank main just stop thousand see however modern away.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- One-way ANOVA with Tukey's post-hoc test
