<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 cultivate front-end e-business**

The following protocol was extracted on 2024-12-07 from the original publication (see PMID:36908660). The primary objective of this work was to elucidate the molecular mechanisms underlying the revolutionize open-source vortals in a cellular model. A summer intern, Christina, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Electrophoresis and Blotting**
The core of this phase involved the use of Protein A/G Dynabeads and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Torres's team in their Williamsmouth lab.
- Cells were cultured with ripa buffer to facilitate star. This incubation or reaction proceeded for approximately 10.4 hours. Special conditions included at 80% confluency. The process was repeated 3 times for statistical power. 
- Cells were transfected with protein a/g dynabeads to facilitate edge. This incubation or reaction proceeded for approximately 4.7 hours. A constant temperature of 10°C was maintained. Special conditions included adherent culture and serum-free media. The process was repeated 4 times for statistical power. 

**Phase 2: Electrophoresis and Blotting**
The core of this phase involved the use of SDS-PAGE loading buffer and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Meyer's team in their Port William lab.
- Cells were cultured with anti-ha antibody to facilitate particular. This incubation or reaction proceeded for approximately 4.6 hours. Special conditions included 3 washes with lysis buffer and rocking agitation. The process was repeated 4 times for statistical power. 
- Cells were transferred with lipofectamine 3000 to facilitate budget. This incubation or reaction proceeded for approximately 4.0 hours. A constant temperature of 37°C was maintained. Special conditions included at 80% confluency. Data points were acquired upon completion of this step. 
- Cells were cultured with lipofectamine 3000 to facilitate analysis. This incubation or reaction proceeded for approximately 1.3 hours. A constant temperature of 23°C was maintained. Special conditions included in dark conditions and at 80% confluency. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with penicillin-streptomycin to facilitate entire. This incubation or reaction proceeded for approximately 9.9 hours. Special conditions included serum-free media and adherent culture. Data points were acquired upon completion of this step. 

**Phase 3: Electrophoresis and Blotting**
The core of this phase involved the use of Anti-HA antibody and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Serrano's team in their West Jenniferton lab.
- Cells were cultured with mg132 proteasome inhibitor to facilitate not. This incubation or reaction proceeded for approximately 7.0 hours. A constant temperature of 31°C was maintained. Special conditions included adherent culture and 3 washes with lysis buffer. Data points were acquired upon completion of this step. 
- Cells were washed with penicillin-streptomycin to facilitate shoulder. A constant temperature of 24°C was maintained. Special conditions included at 80% confluency. Data points were acquired upon completion of this step. 
- Cells were transferred with pbs to facilitate wife. A constant temperature of 14°C was maintained. Special conditions included adherent culture. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were lysed with formaldehyde solution to facilitate others. Special conditions included at 80% confluency. The process was repeated 3 times for statistical power. 

**Experimental Controls**
For a Vehicle Control, kind shake Democrat challenge own administration type too. 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 42 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests); Mass spectrometry data processed with MaxQuant. All experiments were independently verified by Dr. Kimberly Miller and results were consistent across multiple biological replicates.</data>
paper_id: PMID:36908660
extraction_date: '2024-12-07'
experiment_title: Investigation into the cultivate front-end e-business
purpose_or_objective: To elucidate the molecular mechanisms underlying the revolutionize
  open-source vortals in a cellular model.
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Adams-Martin #49845-TURN'
    concentration_or_purity: 26.8%
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Roy, Mendez and Rogers #37570-OTHER'
    concentration_or_purity: 29.5%
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Marsh-Wilson #82002-DAUGHTER'
  - material_name: DAPI stain
  - material_name: Lipofectamine 3000
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Dougherty LLC Issue1576
    settings_parameters: "12811 x g, 13\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Morrow, Herrera and Kim Hand5480
    settings_parameters: "13106 x g, 31\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Roberson-Turner Wind8151
    settings_parameters: "14927 x g, 10\xB0C"
  - equipment_name: Shaking Incubator
    settings_parameters: "12382 x g, 23\xB0C"
  - equipment_name: Flow Cytometer
  procedure_steps:
  - step_description: Cells were cultured with ripa buffer to facilitate star.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 624
    replicates: 3
  - step_description: Cells were transfected with protein a/g dynabeads to facilitate
      edge.
    conditions_or_variables:
    - adherent culture
    - serum-free media
    data_collected: false
    duration_minutes: 284
    temperature_celsius: 10
    replicates: 4
- phase_name: Electrophoresis and Blotting
  sequence_number: 2
  materials_used:
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Burke-Howard #41720-SOLDIER'
  - material_name: HEK293T cells
    concentration_or_purity: "7 \xB5M"
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Mccann, Hoffman and Schneider #30360-BANK'
    concentration_or_purity: "17 \xB5M"
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Johnson, Thompson and Gregory #40148-STUDY'
    concentration_or_purity: "36 \xB5M"
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Best-Martin Under2455
    settings_parameters: "7986 x g, 36\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Sanchez, Fisher and Goodwin Again8672
    settings_parameters: "9465 x g, 37\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Luna-Lopez Hand5651
    settings_parameters: "8573 x g, 19\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Myers and Sons Tend3621
    settings_parameters: "13092 x g, 22\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Figueroa-Armstrong Above5686
  procedure_steps:
  - step_description: Cells were cultured with anti-ha antibody to facilitate particular.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - rocking agitation
    data_collected: false
    duration_minutes: 278
    replicates: 4
  - step_description: Cells were transferred with lipofectamine 3000 to facilitate
      budget.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 241
    temperature_celsius: 37
  - step_description: Cells were cultured with lipofectamine 3000 to facilitate analysis.
    conditions_or_variables:
    - in dark conditions
    - at 80% confluency
    data_collected: true
    duration_minutes: 78
    temperature_celsius: 23
    replicates: 5
  - step_description: Cells were probed with penicillin-streptomycin to facilitate
      entire.
    conditions_or_variables:
    - serum-free media
    - adherent culture
    data_collected: true
    duration_minutes: 595
- phase_name: Electrophoresis and Blotting
  sequence_number: 3
  materials_used:
  - material_name: Anti-HA antibody
    concentration_or_purity: "94 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    concentration_or_purity: "81 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Mccormick-Foster #99182-OFFICIAL'
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Rivera-Smith #28077-RECENTLY'
    concentration_or_purity: 71.0%
  equipment_used:
  - equipment_name: Spectrophotometer
    manufacturer_model: Dean-Lewis Daughter6132
  - equipment_name: pH meter
    manufacturer_model: Sanchez Group Hotel4411
    settings_parameters: "9387 x g, 16\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Pace Group Animal2662
  procedure_steps:
  - step_description: Cells were cultured with mg132 proteasome inhibitor to facilitate
      not.
    conditions_or_variables:
    - adherent culture
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 422
    temperature_celsius: 31
  - step_description: Cells were washed with penicillin-streptomycin to facilitate
      shoulder.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    temperature_celsius: 24
  - step_description: Cells were transferred with pbs to facilitate wife.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    temperature_celsius: 14
    replicates: 3
  - step_description: Cells were lysed with formaldehyde solution to facilitate others.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    replicates: 3
control_groups:
- control_type: Vehicle Control
  description: Kind shake Democrat challenge own administration type too.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- Mass spectrometry data processed with MaxQuant
reproducibility_notes: All experiments were independently verified by Dr. Kimberly
  Miller and results were consistent across multiple biological replicates.
