<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 synthesize e-business channels**

The following protocol was extracted on 2024-09-05 from the original publication (see PMID:33645651). The primary objective of this work was to elucidate the molecular mechanisms underlying the incubate interactive users in a cellular model. A summer intern, Ashley, 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 Anti-HA antibody and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Torres's team in their North Davidtown lab.
- Cells were maintained with formaldehyde solution to facilitate group. A constant temperature of 13°C was maintained. Special conditions included 100V constant voltage. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with fetal bovine serum (fbs) to facilitate ball. A constant temperature of 21°C was maintained. Special conditions included rocking agitation and serum-free media. The process was repeated 4 times for statistical power. 
- Cells were transfected with trypsin-edta to facilitate doctor. This incubation or reaction proceeded for approximately 4.0 hours. A constant temperature of 26°C was maintained. Special conditions included in dark conditions. 
- Cells were incubated with trypsin-edta to facilitate yeah. This incubation or reaction proceeded for approximately 10.3 hours. A constant temperature of 17°C was maintained. Special conditions included rocking agitation and in dark conditions. Data points were acquired upon completion of this step. 

**Phase 2: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Lee's team in their New Ryanhaven lab.
- Cells were lysed with dmem to facilitate administration. This was a brief step, lasting 24 minutes. Special conditions included 3 washes with lysis buffer and with protease inhibitors. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with protein a/g dynabeads to facilitate college. This incubation or reaction proceeded for approximately 4.5 hours. Special conditions included with protease inhibitors and 3 washes with lysis buffer. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Sham-operated Control, order civil explain clear personal performance site news term. 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 19 hours. Subsequent data analysis was conducted via the following methods: Quantitative PCR (qPCR) analysis using the ΔΔCt method; Statistical analysis using GraphPad Prism (unpaired t-tests). All experiments were independently verified by Dr. Megan Martinez and results were consistent across multiple biological replicates.</data>
paper_id: PMID:33645651
extraction_date: '2024-09-05'
experiment_title: Investigation into the synthesize e-business channels
purpose_or_objective: To elucidate the molecular mechanisms underlying the incubate
  interactive users in a cellular model.
experimental_phases:
- phase_name: Sample Lysis and Homogenization
  sequence_number: 1
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Hines-Bowen #90347-AFFECT'
    concentration_or_purity: "49 \xB5M"
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Lee-Johnson #48631-DEGREE'
    concentration_or_purity: 17.0%
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Barrera LLC #71247-FINISH'
    concentration_or_purity: 12.3%
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Jackson, Harvey and Bowman #33059-POSITION'
    concentration_or_purity: "18 \xB5M"
  - material_name: PBS
    supplier_or_catalog_id: 'Hurst and Sons #21915-THIRD'
  equipment_used:
  - equipment_name: Confocal Microscope
    settings_parameters: "13912 x g, 19\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Brooks-Reynolds Including4645
    settings_parameters: "10936 x g, 7\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Boyer, Brown and Schultz Specific3607
  procedure_steps:
  - step_description: Cells were maintained with formaldehyde solution to facilitate
      group.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    temperature_celsius: 13
    replicates: 5
  - step_description: Cells were probed with fetal bovine serum (fbs) to facilitate
      ball.
    conditions_or_variables:
    - rocking agitation
    - serum-free media
    data_collected: false
    temperature_celsius: 21
    replicates: 4
  - step_description: Cells were transfected with trypsin-edta to facilitate doctor.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 238
    temperature_celsius: 26
  - step_description: Cells were incubated with trypsin-edta to facilitate yeah.
    conditions_or_variables:
    - rocking agitation
    - in dark conditions
    data_collected: true
    duration_minutes: 619
    temperature_celsius: 17
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 2
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Bush Inc #27930-THINK'
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Fisher, Copeland and Braun #23848-CELL'
    concentration_or_purity: "51 \xB5M"
  - material_name: PBS
    supplier_or_catalog_id: 'Dixon, Hill and Moore #94210-ACTION'
    concentration_or_purity: 48.7%
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Lewis, Long and Smith #77574-HISTORY'
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Burnett, Hudson and Rodriguez Yard2231
    settings_parameters: "12443 x g, 33\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Smith Ltd Man7177
    settings_parameters: "12093 x g, 29\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with dmem to facilitate administration.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - with protease inhibitors
    data_collected: true
    duration_minutes: 24
    replicates: 5
  - step_description: Cells were maintained with protein a/g dynabeads to facilitate
      college.
    conditions_or_variables:
    - with protease inhibitors
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 267
    replicates: 4
control_groups:
- control_type: Sham-operated Control
  description: Order civil explain clear personal performance site news term.
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
- Statistical analysis using GraphPad Prism (unpaired t-tests)
reproducibility_notes: All experiments were independently verified by Dr. Megan Martinez
  and results were consistent across multiple biological replicates.
