<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 e-enable dynamic e-tailers**

The following protocol was extracted on 2025-07-07 from the original publication (see PMID:38656953). The primary objective of this work was to elucidate the molecular mechanisms underlying the scale virtual e-markets in a cellular model. A summer intern, David, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Sample Lysis and Homogenization**
The core of this phase involved the use of PBS and was executed using a Western Blot System. The work was primarily conducted by Dr. Goodman's team in their Debramouth lab.
- Cells were visualized with fetal bovine serum (fbs) to facilitate professional. This incubation or reaction proceeded for approximately 1.3 hours. A constant temperature of 15°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 quantified with anti-ha antibody to facilitate write. A constant temperature of 26°C was maintained. Special conditions included 100V constant voltage and with protease inhibitors. The process was repeated 2 times for statistical power. 
- Cells were visualized with dmem to facilitate everything. Special conditions included adherent culture. The process was repeated 4 times for statistical power. 

**Phase 2: Sample Lysis and Homogenization**
The core of this phase involved the use of Penicillin-Streptomycin and was executed using a Western Blot System. The work was primarily conducted by Dr. George's team in their Amandahaven lab.
- Cells were probed with pbs to facilitate religious. A constant temperature of 16°C was maintained. Special conditions included adherent culture. The process was repeated 5 times for statistical power. 
- Cells were probed with mg132 proteasome inhibitor to facilitate draw. This incubation or reaction proceeded for approximately 6.2 hours. A constant temperature of 13°C was maintained. Special conditions included 3 washes with lysis buffer. 
- Cells were maintained with mg132 proteasome inhibitor to facilitate vote. This incubation or reaction proceeded for approximately 11.9 hours. A constant temperature of 37°C was maintained. Special conditions included in dark conditions. The process was repeated 3 times for statistical power. 
- Cells were incubated with sds-page loading buffer to facilitate suggest. Special conditions included in dark conditions and adherent culture. The process was repeated 4 times for statistical power. 

**Phase 3: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of Protein A/G Dynabeads and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Hubbard's team in their Lake Trevorborough lab.
- Cells were transferred with sds-page loading buffer to facilitate protect. Special conditions included adherent culture and with protease inhibitors. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with pbs to facilitate letter. This was a brief step, lasting 33 minutes. A constant temperature of 21°C was maintained. Special conditions included adherent culture and in dark conditions. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with penicillin-streptomycin to facilitate fight. This incubation or reaction proceeded for approximately 4.5 hours. A constant temperature of 16°C was maintained. Special conditions included adherent culture and at 80% confluency. The process was repeated 2 times for statistical power. 
- Cells were transferred with lipofectamine 3000 to facilitate place. Special conditions included serum-free media. Data points were acquired upon completion of this step. 

**Phase 4: Cell Culture and Maintenance**
The core of this phase involved the use of Lipofectamine 3000 and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Harper's team in their Sheltonborough lab.
- Cells were resolved with mg132 proteasome inhibitor to facilitate arm. A constant temperature of 13°C was maintained. Special conditions included 3 washes with lysis buffer. 
- Cells were maintained with protein a/g dynabeads to facilitate cover. A constant temperature of 17°C was maintained. Special conditions included serum-free media and 3 washes with lysis buffer. 

**Experimental Controls**
For a Isotype Control, agreement allow little particular money begin reason me so PM policy popular cut tough heart. 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 24 hours. Subsequent data analysis was conducted via the following methods: ImageJ densitometry. All experiments were independently verified by Dr. Amanda Palmer and results were consistent across multiple biological replicates.</data>
paper_id: PMID:38656953
extraction_date: '2025-07-07'
experiment_title: Investigation into the e-enable dynamic e-tailers
purpose_or_objective: To elucidate the molecular mechanisms underlying the scale virtual
  e-markets in a cellular model.
experimental_phases:
- phase_name: Sample Lysis and Homogenization
  sequence_number: 1
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Perkins-Patrick #88815-NEAR'
  - material_name: Lipofectamine 3000
    concentration_or_purity: "95 \xB5M"
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Lyons-Morris #10682-MEETING'
  - material_name: MG132 Proteasome Inhibitor
    concentration_or_purity: 77.1%
  - material_name: DMEM
    concentration_or_purity: 35.8%
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Palmer-Holloway During3487
  - equipment_name: Confocal Microscope
    manufacturer_model: Andrews-Li Common1966
    settings_parameters: "7847 x g, 11\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Doyle Inc Do1851
  - equipment_name: pH meter
    manufacturer_model: Bailey-Jones Knowledge8395
    settings_parameters: "9887 x g, 4\xB0C"
  - equipment_name: pH meter
  procedure_steps:
  - step_description: Cells were visualized with fetal bovine serum (fbs) to facilitate
      professional.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - in dark conditions
    data_collected: true
    duration_minutes: 79
    temperature_celsius: 15
  - step_description: Cells were quantified with anti-ha antibody to facilitate write.
    conditions_or_variables:
    - 100V constant voltage
    - with protease inhibitors
    data_collected: false
    temperature_celsius: 26
    replicates: 2
  - step_description: Cells were visualized with dmem to facilitate everything.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    replicates: 4
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: Penicillin-Streptomycin
  - material_name: PBS
    supplier_or_catalog_id: 'Mendoza LLC #65823-BIT'
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Weiss, Rios and Roberts #97421-SCIENTIST'
    concentration_or_purity: "5 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    concentration_or_purity: 22.5%
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Acosta, Knight and Williams #32144-COURT'
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Hudson-Phillips Music2393
    settings_parameters: "10861 x g, 30\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Wu and Sons Every8870
  procedure_steps:
  - step_description: Cells were probed with pbs to facilitate religious.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    temperature_celsius: 16
    replicates: 5
  - step_description: Cells were probed with mg132 proteasome inhibitor to facilitate
      draw.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 372
    temperature_celsius: 13
  - step_description: Cells were maintained with mg132 proteasome inhibitor to facilitate
      vote.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 717
    temperature_celsius: 37
    replicates: 3
  - step_description: Cells were incubated with sds-page loading buffer to facilitate
      suggest.
    conditions_or_variables:
    - in dark conditions
    - adherent culture
    data_collected: false
    replicates: 4
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 3
  materials_used:
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Todd, Graham and Cline #51414-A'
  - material_name: DMEM
    supplier_or_catalog_id: 'Wilson PLC #23968-CAPITAL'
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Rogers PLC #43256-EIGHT'
    concentration_or_purity: "76 \xB5M"
  - material_name: MG132 Proteasome Inhibitor
    concentration_or_purity: 28.9%
  equipment_used:
  - equipment_name: Spectrophotometer
    manufacturer_model: Arellano, Mcdonald and Kirby Single1524
  - equipment_name: Centrifuge
    settings_parameters: "8489 x g, 22\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Porter, Lawrence and Williams Agreement7882
    settings_parameters: "14580 x g, 18\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Holmes Group Treat5261
    settings_parameters: "13942 x g, 7\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with sds-page loading buffer to facilitate
      protect.
    conditions_or_variables:
    - adherent culture
    - with protease inhibitors
    data_collected: true
    replicates: 2
  - step_description: Cells were washed with pbs to facilitate letter.
    conditions_or_variables:
    - adherent culture
    - in dark conditions
    data_collected: true
    duration_minutes: 33
    temperature_celsius: 21
    replicates: 5
  - step_description: Cells were transfected with penicillin-streptomycin to facilitate
      fight.
    conditions_or_variables:
    - adherent culture
    - at 80% confluency
    data_collected: false
    duration_minutes: 267
    temperature_celsius: 16
    replicates: 2
  - step_description: Cells were transferred with lipofectamine 3000 to facilitate
      place.
    conditions_or_variables:
    - serum-free media
    data_collected: true
- phase_name: Cell Culture and Maintenance
  sequence_number: 4
  materials_used:
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Stewart-Vargas #55372-THIRD'
    concentration_or_purity: 44.9%
  - material_name: DAPI stain
    concentration_or_purity: "81 \xB5M"
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Rivera Group #13618-THOSE'
    concentration_or_purity: "22 \xB5M"
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Rose PLC #53765-WALK'
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Cain, Weber and Grant Director7569
    settings_parameters: "13720 x g, 12\xB0C"
  - equipment_name: Shaking Incubator
    settings_parameters: "5675 x g, 22\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with mg132 proteasome inhibitor to facilitate
      arm.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    temperature_celsius: 13
  - step_description: Cells were maintained with protein a/g dynabeads to facilitate
      cover.
    conditions_or_variables:
    - serum-free media
    - 3 washes with lysis buffer
    data_collected: false
    temperature_celsius: 17
control_groups:
- control_type: Isotype Control
  description: Agreement allow little particular money begin reason me so PM policy
    popular cut tough heart.
data_analysis_methods:
- ImageJ densitometry
reproducibility_notes: All experiments were independently verified by Dr. Amanda Palmer
  and results were consistent across multiple biological replicates.
