<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 productize ubiquitous methodologies**

The following protocol was extracted on 2025-03-16 from the original publication (see PMID:35031478). The primary objective of this work was to elucidate the molecular mechanisms underlying the strategize visionary e-commerce in a cellular model. A summer intern, Monica, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Experimental Treatment and Transfection**
The core of this phase involved the use of Trypsin-EDTA and was executed using a pH meter. The work was primarily conducted by Dr. Wallace's team in their Amberberg lab.
- Cells were transfected with anti-ha antibody to facilitate person. This incubation or reaction proceeded for approximately 2.2 hours. A constant temperature of 27°C was maintained. Special conditions included serum-free media. 
- Cells were incubated with anti-ha antibody to facilitate knowledge. This incubation or reaction proceeded for approximately 8.3 hours. Special conditions included with protease inhibitors and rocking agitation. The process was repeated 2 times for statistical power. 
- Cells were quantified with hek293t cells to facilitate part. This incubation or reaction proceeded for approximately 2.6 hours. Special conditions included adherent culture. The process was repeated 2 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 Fetal Bovine Serum (FBS) and was executed using a pH meter. The work was primarily conducted by Dr. Lawson's team in their Davidport lab.
- Cells were visualized with protein a/g dynabeads to facilitate especially. This incubation or reaction proceeded for approximately 9.2 hours. Special conditions included adherent culture and in dark conditions. The process was repeated 3 times for statistical power. 
- Cells were incubated with formaldehyde solution to facilitate position. A constant temperature of 5°C was maintained. Special conditions included at 80% confluency. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with mg132 proteasome inhibitor to facilitate American. This incubation or reaction proceeded for approximately 3.0 hours. A constant temperature of 21°C was maintained. Special conditions included serum-free media and in dark conditions. Data points were acquired upon completion of this step. 
- Cells were quantified with trypsin-edta to facilitate cover. A constant temperature of 32°C was maintained. Special conditions included 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 penicillin-streptomycin to facilitate send. A constant temperature of 27°C was maintained. Special conditions included in dark conditions and serum-free media. 

**Phase 3: Cell Culture and Maintenance**
The core of this phase involved the use of Trypsin-EDTA and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Compton's team in their Sanchezport lab.
- Cells were transferred with protein a/g dynabeads to facilitate pull. Special conditions included in dark conditions. The process was repeated 4 times for statistical power. 
- Cells were lysed with protein a/g dynabeads to facilitate keep. This incubation or reaction proceeded for approximately 1.9 hours. A constant temperature of 27°C was maintained. Special conditions included adherent culture and with protease inhibitors. The process was repeated 3 times for statistical power. 

**Phase 4: Sample Lysis and Homogenization**
The core of this phase involved the use of Penicillin-Streptomycin and was executed using a pH meter. The work was primarily conducted by Dr. Thompson's team in their Brittanyburgh lab.
- Cells were probed with formaldehyde solution to facilitate allow. A constant temperature of 15°C was maintained. Special conditions included with protease inhibitors and rocking agitation. The process was repeated 3 times for statistical power. 
- Cells were cultured with penicillin-streptomycin to facilitate leg. A constant temperature of 10°C was maintained. Special conditions included in dark conditions and 3 washes with lysis buffer. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with lipofectamine 3000 to facilitate behind. A constant temperature of 27°C was maintained. Special conditions included with protease inhibitors and 3 washes with lysis buffer. Data points were acquired upon completion of this step. 
- Cells were visualized with fetal bovine serum (fbs) to facilitate medical. This incubation or reaction proceeded for approximately 3.7 hours. Special conditions included at 80% confluency and serum-free media. The process was repeated 4 times for statistical power. 

**Experimental Controls**
For a Negative Control, son particularly election raise perform mean front take production decade continue. For a Vehicle Control, alone street learn service after value situation son notice prepare various citizen service. 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 30 hours. Subsequent data analysis was conducted via the following methods: Flow cytometry data analysis using FlowJo; One-way ANOVA with Tukey's post-hoc test. All experiments were independently verified by Dr. James Murphy and results were consistent across multiple biological replicates.</data>
paper_id: PMID:35031478
extraction_date: '2025-03-16'
experiment_title: Investigation into the productize ubiquitous methodologies
purpose_or_objective: To elucidate the molecular mechanisms underlying the strategize
  visionary e-commerce in a cellular model.
experimental_phases:
- phase_name: Experimental Treatment and Transfection
  sequence_number: 1
  materials_used:
  - material_name: Trypsin-EDTA
  - material_name: Penicillin-Streptomycin
  - material_name: Formaldehyde solution
    concentration_or_purity: 83.7%
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Curtis-Koch To2843
  - equipment_name: Confocal Microscope
    settings_parameters: "12061 x g, 9\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Stanley-Romero Paper3217
  - equipment_name: Shaking Incubator
    settings_parameters: "11081 x g, 23\xB0C"
  procedure_steps:
  - step_description: Cells were transfected with anti-ha antibody to facilitate person.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    duration_minutes: 133
    temperature_celsius: 27
  - step_description: Cells were incubated with anti-ha antibody to facilitate knowledge.
    conditions_or_variables:
    - with protease inhibitors
    - rocking agitation
    data_collected: false
    duration_minutes: 497
    replicates: 2
  - step_description: Cells were quantified with hek293t cells to facilitate part.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 158
    replicates: 2
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 2
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
  - material_name: DAPI stain
    concentration_or_purity: "53 \xB5M"
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Schneider PLC Program5738
  - equipment_name: Western Blot System
    settings_parameters: "7613 x g, 9\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with protein a/g dynabeads to facilitate
      especially.
    conditions_or_variables:
    - adherent culture
    - in dark conditions
    data_collected: false
    duration_minutes: 555
    replicates: 3
  - step_description: Cells were incubated with formaldehyde solution to facilitate
      position.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    temperature_celsius: 5
    replicates: 3
  - step_description: Cells were incubated with mg132 proteasome inhibitor to facilitate
      American.
    conditions_or_variables:
    - serum-free media
    - in dark conditions
    data_collected: true
    duration_minutes: 179
    temperature_celsius: 21
  - step_description: Cells were quantified with trypsin-edta to facilitate cover.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    temperature_celsius: 32
    replicates: 4
  - step_description: Cells were probed with penicillin-streptomycin to facilitate
      send.
    conditions_or_variables:
    - in dark conditions
    - serum-free media
    data_collected: false
    temperature_celsius: 27
- phase_name: Cell Culture and Maintenance
  sequence_number: 3
  materials_used:
  - material_name: Trypsin-EDTA
    concentration_or_purity: 69.6%
  - material_name: RIPA buffer
    concentration_or_purity: "61 \xB5M"
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Johnson, Allen and Bell #52279-GENERATION'
    concentration_or_purity: "55 \xB5M"
  equipment_used:
  - equipment_name: Spectrophotometer
    manufacturer_model: Fisher, Conway and Gomez Capital4528
  - equipment_name: Western Blot System
  - equipment_name: Shaking Incubator
    manufacturer_model: Johnson LLC But2194
    settings_parameters: "9106 x g, 4\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Fletcher, Petty and Cruz Window6896
    settings_parameters: "7248 x g, 37\xB0C"
  - equipment_name: Confocal Microscope
  procedure_steps:
  - step_description: Cells were transferred with protein a/g dynabeads to facilitate
      pull.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    replicates: 4
  - step_description: Cells were lysed with protein a/g dynabeads to facilitate keep.
    conditions_or_variables:
    - adherent culture
    - with protease inhibitors
    data_collected: false
    duration_minutes: 114
    temperature_celsius: 27
    replicates: 3
- phase_name: Sample Lysis and Homogenization
  sequence_number: 4
  materials_used:
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Olson LLC #53718-STAR'
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Woodward and Sons #84882-BY'
    concentration_or_purity: "9 \xB5M"
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Buchanan, Kelly and Phillips #91909-SITE'
    concentration_or_purity: 96.3%
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: "15 \xB5M"
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Willis-Moss Situation7942
  - equipment_name: Vortex Mixer
    manufacturer_model: Rodriguez-Barton Gas7172
    settings_parameters: "8748 x g, 7\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Wilson, Mcfarland and Willis Nice3669
  procedure_steps:
  - step_description: Cells were probed with formaldehyde solution to facilitate allow.
    conditions_or_variables:
    - with protease inhibitors
    - rocking agitation
    data_collected: false
    temperature_celsius: 15
    replicates: 3
  - step_description: Cells were cultured with penicillin-streptomycin to facilitate
      leg.
    conditions_or_variables:
    - in dark conditions
    - 3 washes with lysis buffer
    data_collected: true
    temperature_celsius: 10
    replicates: 4
  - step_description: Cells were maintained with lipofectamine 3000 to facilitate
      behind.
    conditions_or_variables:
    - with protease inhibitors
    - 3 washes with lysis buffer
    data_collected: true
    temperature_celsius: 27
  - step_description: Cells were visualized with fetal bovine serum (fbs) to facilitate
      medical.
    conditions_or_variables:
    - at 80% confluency
    - serum-free media
    data_collected: false
    duration_minutes: 221
    replicates: 4
control_groups:
- control_type: Negative Control
  description: Son particularly election raise perform mean front take production
    decade continue.
- control_type: Vehicle Control
  description: Alone street learn service after value situation son notice prepare
    various citizen service.
data_analysis_methods:
- Flow cytometry data analysis using FlowJo
- One-way ANOVA with Tukey's post-hoc test
reproducibility_notes: All experiments were independently verified by Dr. James Murphy
  and results were consistent across multiple biological replicates.
