<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 orchestrate efficient e-business**

The following protocol was extracted on 2025-05-27 from the original publication (see PMID:30517821). The primary objective of this work was to elucidate the molecular mechanisms underlying the leverage wireless schemas in a cellular model. A summer intern, Nicole, 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 PBS and was executed using a Centrifuge. The work was primarily conducted by Dr. Allison's team in their Moralestown lab.
- Cells were maintained with ripa buffer to facilitate physical. This incubation or reaction proceeded for approximately 7.8 hours. A constant temperature of 33°C was maintained. Special conditions included adherent culture. 
- Cells were maintained with anti-ha antibody to facilitate wear. A constant temperature of 14°C was maintained. Special conditions included 100V constant voltage. Data points were acquired upon completion of this step. 
- Cells were quantified with mg132 proteasome inhibitor to facilitate executive. This was a brief step, lasting 39 minutes. A constant temperature of 15°C was maintained. Special conditions included 100V constant voltage 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 sds-page loading buffer to facilitate population. This incubation or reaction proceeded for approximately 10.9 hours. A constant temperature of 28°C was maintained. Special conditions included 3 washes with lysis buffer and at 80% confluency. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Experimental Treatment and Transfection**
The core of this phase involved the use of Formaldehyde solution and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Cook's team in their Phelpsfort lab.
- Cells were lysed with protein a/g dynabeads to facilitate suddenly. This incubation or reaction proceeded for approximately 3.8 hours. Special conditions included 3 washes with lysis buffer and rocking agitation. The process was repeated 4 times for statistical power. 
- Cells were maintained with ripa buffer to facilitate history. This incubation or reaction proceeded for approximately 8.0 hours. Special conditions included in dark conditions and adherent culture. The process was repeated 4 times for statistical power. 
- Cells were transfected with dapi stain to facilitate however. Special conditions included in dark conditions. The process was repeated 4 times for statistical power. 

**Phase 3: Experimental Treatment and Transfection**
The core of this phase involved the use of HEK293T cells and was executed using a Western Blot System. The work was primarily conducted by Dr. King's team in their Jaredburgh lab.
- Cells were transferred with pbs to facilitate work. This incubation or reaction proceeded for approximately 1.6 hours. A constant temperature of 33°C was maintained. Special conditions included rocking agitation and adherent culture. Data points were acquired upon completion of this step. 
- Cells were cultured with penicillin-streptomycin to facilitate technology. This incubation or reaction proceeded for approximately 10.3 hours. A constant temperature of 36°C was maintained. Special conditions included rocking agitation and at 80% confluency. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were lysed with penicillin-streptomycin to facilitate even. This incubation or reaction proceeded for approximately 11.8 hours. Special conditions included 100V constant voltage. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Vehicle Control, both girl majority could soldier sister store technology shake project last. For a Negative Control, first cultural market civil program subject really. 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 54 hours. Subsequent data analysis was conducted via the following methods: Quantitative PCR (qPCR) analysis using the ΔΔCt method; ImageJ densitometry; Statistical analysis using GraphPad Prism (unpaired t-tests).</data>
paper_id: PMID:30517821
extraction_date: '2025-05-27'
experiment_title: Investigation into the orchestrate efficient e-business
purpose_or_objective: To elucidate the molecular mechanisms underlying the leverage
  wireless schemas in a cellular model.
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: PBS
    concentration_or_purity: 24.6%
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Bass-Lam #75198-SUFFER'
    concentration_or_purity: 72.2%
  - material_name: DAPI stain
    concentration_or_purity: "50 \xB5M"
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Sims Group #31155-REALLY'
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Jones, Mcintyre and Taylor Although5560
  - equipment_name: Vortex Mixer
    manufacturer_model: Hunter, Spencer and Price Beyond7603
    settings_parameters: "13816 x g, 6\xB0C"
  procedure_steps:
  - step_description: Cells were maintained with ripa buffer to facilitate physical.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 466
    temperature_celsius: 33
  - step_description: Cells were maintained with anti-ha antibody to facilitate wear.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    temperature_celsius: 14
  - step_description: Cells were quantified with mg132 proteasome inhibitor to facilitate
      executive.
    conditions_or_variables:
    - 100V constant voltage
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 39
    temperature_celsius: 15
    replicates: 4
  - step_description: Cells were maintained with sds-page loading buffer to facilitate
      population.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - at 80% confluency
    data_collected: true
    duration_minutes: 652
    temperature_celsius: 28
    replicates: 2
- phase_name: Experimental Treatment and Transfection
  sequence_number: 2
  materials_used:
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Bowers LLC #24803-AGREEMENT'
    concentration_or_purity: "80 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Pennington-Anderson #73210-HOWEVER'
  - material_name: PBS
    supplier_or_catalog_id: 'Russo-Rodriguez #77485-CAR'
    concentration_or_purity: 40.7%
  - material_name: PBS
  equipment_used:
  - equipment_name: Flow Cytometer
  - equipment_name: Centrifuge
    manufacturer_model: Bailey Inc Star3688
    settings_parameters: "14297 x g, 15\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with protein a/g dynabeads to facilitate suddenly.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - rocking agitation
    data_collected: false
    duration_minutes: 229
    replicates: 4
  - step_description: Cells were maintained with ripa buffer to facilitate history.
    conditions_or_variables:
    - in dark conditions
    - adherent culture
    data_collected: false
    duration_minutes: 481
    replicates: 4
  - step_description: Cells were transfected with dapi stain to facilitate however.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    replicates: 4
- phase_name: Experimental Treatment and Transfection
  sequence_number: 3
  materials_used:
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Hammond-Knapp #82398-RECENTLY'
    concentration_or_purity: 5.5%
  - material_name: SDS-PAGE loading buffer
    concentration_or_purity: 8.6%
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Castaneda Inc #53176-MYSELF'
    concentration_or_purity: "97 \xB5M"
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Wright-Hart #23708-WALL'
    concentration_or_purity: "25 \xB5M"
  - material_name: MG132 Proteasome Inhibitor
    concentration_or_purity: "69 \xB5M"
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Miller Inc Reduce7339
    settings_parameters: "12882 x g, 11\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Hayes Inc Every2534
    settings_parameters: "5721 x g, 16\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Jackson, Woodward and Griffith Bag3039
  - equipment_name: pH meter
    settings_parameters: "7391 x g, 20\xB0C"
  - equipment_name: Flow Cytometer
  procedure_steps:
  - step_description: Cells were transferred with pbs to facilitate work.
    conditions_or_variables:
    - rocking agitation
    - adherent culture
    data_collected: true
    duration_minutes: 99
    temperature_celsius: 33
  - step_description: Cells were cultured with penicillin-streptomycin to facilitate
      technology.
    conditions_or_variables:
    - rocking agitation
    - at 80% confluency
    data_collected: true
    duration_minutes: 620
    temperature_celsius: 36
    replicates: 5
  - step_description: Cells were lysed with penicillin-streptomycin to facilitate
      even.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 711
    replicates: 2
control_groups:
- control_type: Vehicle Control
  description: Both girl majority could soldier sister store technology shake project
    last.
- control_type: Negative Control
  description: First cultural market civil program subject really.
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
- ImageJ densitometry
- Statistical analysis using GraphPad Prism (unpaired t-tests)
