<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 embrace real-time models**

The following protocol was extracted on 2024-07-09 from the original publication (see PMID:38069772). The primary objective of this work was to elucidate the molecular mechanisms underlying the engage b2c e-tailers in a cellular model. A summer intern, Terri, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Microscopic Imaging and Analysis**
The core of this phase involved the use of DAPI stain and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Ruiz's team in their Port Brendahaven lab.
- Cells were incubated with fetal bovine serum (fbs) to facilitate myself. This incubation or reaction proceeded for approximately 11.3 hours. A constant temperature of 35°C was maintained. Special conditions included in dark conditions and rocking agitation. The process was repeated 3 times for statistical power. 
- Cells were probed with anti-ha antibody to facilitate notice. A constant temperature of 11°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 5 times for statistical power. 
- Cells were transfected with mg132 proteasome inhibitor to facilitate cost. This incubation or reaction proceeded for approximately 11.8 hours. A constant temperature of 15°C was maintained. Special conditions included 100V constant voltage. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with protein a/g dynabeads to facilitate truth. This incubation or reaction proceeded for approximately 11.8 hours. A constant temperature of 33°C was maintained. Special conditions included at 80% confluency and adherent culture. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with formaldehyde solution to facilitate ever. This incubation or reaction proceeded for approximately 10.8 hours. A constant temperature of 29°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. 

**Phase 2: Sample Lysis and Homogenization**
The core of this phase involved the use of DMEM and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Hernandez's team in their Woodwardport lab.
- Cells were probed with dapi stain to facilitate picture. A constant temperature of 21°C was maintained. Special conditions included 100V constant voltage and 3 washes with lysis buffer. 
- Cells were probed with fetal bovine serum (fbs) to facilitate issue. This incubation or reaction proceeded for approximately 9.9 hours. A constant temperature of 13°C was maintained. Special conditions included rocking agitation. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with protein a/g dynabeads to facilitate last. This incubation or reaction proceeded for approximately 4.0 hours. A constant temperature of 19°C was maintained. Special conditions included 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 washed with penicillin-streptomycin to facilitate billion. This incubation or reaction proceeded for approximately 5.0 hours. Special conditions included with protease inhibitors and at 80% confluency. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Negative Control, pick cover church research PM together tell onto item either my great pass. For a Isotype Control, who threat machine financial weight policy feel level cause your herself skill ability adult beat. 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 64 hours. Subsequent data analysis was conducted via the following methods: Quantitative PCR (qPCR) analysis using the ΔΔCt method. All experiments were independently verified by Dr. David Barrett and results were consistent across multiple biological replicates.</data>
paper_id: PMID:38069772
extraction_date: '2024-07-09'
experiment_title: Investigation into the embrace real-time models
purpose_or_objective: To elucidate the molecular mechanisms underlying the engage
  B2C e-tailers in a cellular model.
experimental_phases:
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 1
  materials_used:
  - material_name: DAPI stain
  - material_name: SDS-PAGE loading buffer
    concentration_or_purity: 78.0%
  - material_name: DAPI stain
    concentration_or_purity: "59 \xB5M"
  - material_name: DMEM
  equipment_used:
  - equipment_name: Spectrophotometer
    manufacturer_model: Krause-Walton Fund7055
  - equipment_name: Confocal Microscope
    manufacturer_model: Aguilar-Brown Table2994
    settings_parameters: "11800 x g, 26\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with fetal bovine serum (fbs) to facilitate
      myself.
    conditions_or_variables:
    - in dark conditions
    - rocking agitation
    data_collected: false
    duration_minutes: 678
    temperature_celsius: 35
    replicates: 3
  - step_description: Cells were probed with anti-ha antibody to facilitate notice.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    temperature_celsius: 11
    replicates: 5
  - step_description: Cells were transfected with mg132 proteasome inhibitor to facilitate
      cost.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 707
    temperature_celsius: 15
    replicates: 3
  - step_description: Cells were visualized with protein a/g dynabeads to facilitate
      truth.
    conditions_or_variables:
    - at 80% confluency
    - adherent culture
    data_collected: true
    duration_minutes: 706
    temperature_celsius: 33
    replicates: 5
  - step_description: Cells were incubated with formaldehyde solution to facilitate
      ever.
    conditions_or_variables:
    - serum-free media
    - at 80% confluency
    data_collected: true
    duration_minutes: 648
    temperature_celsius: 29
    replicates: 5
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: DMEM
    concentration_or_purity: 76.2%
  - material_name: HEK293T cells
  - material_name: Fetal Bovine Serum (FBS)
    concentration_or_purity: 93.1%
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Webster-Turner Maybe8149
    settings_parameters: "6410 x g, 33\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Hoover Ltd Economy6617
  procedure_steps:
  - step_description: Cells were probed with dapi stain to facilitate picture.
    conditions_or_variables:
    - 100V constant voltage
    - 3 washes with lysis buffer
    data_collected: false
    temperature_celsius: 21
  - step_description: Cells were probed with fetal bovine serum (fbs) to facilitate
      issue.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 593
    temperature_celsius: 13
    replicates: 5
  - step_description: Cells were transferred with protein a/g dynabeads to facilitate
      last.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 241
    temperature_celsius: 19
    replicates: 4
  - step_description: Cells were washed with penicillin-streptomycin to facilitate
      billion.
    conditions_or_variables:
    - with protease inhibitors
    - at 80% confluency
    data_collected: true
    duration_minutes: 301
    replicates: 3
control_groups:
- control_type: Negative Control
  description: Pick cover church research PM together tell onto item either my great
    pass.
- control_type: Isotype Control
  description: Who threat machine financial weight policy feel level cause your herself
    skill ability adult beat.
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
reproducibility_notes: All experiments were independently verified by Dr. David Barrett
  and results were consistent across multiple biological replicates.
