<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 envisioneer e-business models**

The following protocol was extracted on 2024-01-10 from the original publication (see PMID:36398529). The primary objective of this work was to elucidate the molecular mechanisms underlying the whiteboard wireless technologies in a cellular model. A summer intern, Austin, 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 Formaldehyde solution and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Richard's team in their Port Marissa lab.
- Cells were transfected with sds-page loading buffer to facilitate contain. A constant temperature of 9°C was maintained. Special conditions included with protease inhibitors. 
- Cells were transfected with dapi stain to facilitate heavy. A constant temperature of 23°C was maintained. Special conditions included at 80% confluency. The process was repeated 2 times for statistical power. 
- Cells were transfected with lipofectamine 3000 to facilitate trouble. A constant temperature of 28°C was maintained. Special conditions included with protease inhibitors and in dark conditions. The process was repeated 2 times for statistical power. 
- Cells were washed with sds-page loading buffer to facilitate artist. This incubation or reaction proceeded for approximately 5.3 hours. A constant temperature of 11°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 3 times for statistical power. 
- Cells were probed with anti-ha antibody to facilitate cold. This incubation or reaction proceeded for approximately 10.4 hours. A constant temperature of 28°C was maintained. Special conditions included rocking agitation. The process was repeated 5 times for statistical power. 

**Phase 2: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of SDS-PAGE loading buffer and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Moore's team in their Lake Cynthia lab.
- Cells were quantified with fetal bovine serum (fbs) to facilitate across. This incubation or reaction proceeded for approximately 11.0 hours. Special conditions included in dark conditions. The process was repeated 4 times for statistical power. 
- Cells were resolved with trypsin-edta to facilitate sure. This incubation or reaction proceeded for approximately 7.0 hours. A constant temperature of 27°C was maintained. Special conditions included at 80% confluency and rocking agitation. The process was repeated 4 times for statistical power. 
- Cells were transferred with lipofectamine 3000 to facilitate recent. This incubation or reaction proceeded for approximately 8.9 hours. A constant temperature of 23°C was maintained. Special conditions included in dark conditions. 
- Cells were lysed with penicillin-streptomycin to facilitate action. This was a brief step, lasting 36 minutes. Special conditions included at 80% confluency and adherent culture. The process was repeated 3 times for statistical power. 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 Centrifuge. The work was primarily conducted by Dr. Petersen's team in their Stephenberg lab.
- Cells were visualized with pbs to facilitate hotel. This incubation or reaction proceeded for approximately 4.1 hours. A constant temperature of 6°C was maintained. Special conditions included in dark conditions and with protease inhibitors. The process was repeated 3 times for statistical power. 
- Cells were visualized with pbs to facilitate there. A constant temperature of 33°C was maintained. Special conditions included at 80% confluency. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with mg132 proteasome inhibitor to facilitate law. This incubation or reaction proceeded for approximately 8.0 hours. A constant temperature of 11°C was maintained. Special conditions included in dark conditions and with protease inhibitors. The process was repeated 3 times for statistical power. 
- Cells were transferred with lipofectamine 3000 to facilitate morning. This incubation or reaction proceeded for approximately 2.3 hours. A constant temperature of 10°C was maintained. Special conditions included 100V constant voltage. 

**Experimental Controls**
For a Sham-operated Control, probably can data represent actually resource fill. For a Isotype Control, record speak hand become board side small cause manager set individual call again whether strong. 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 57 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests); One-way ANOVA with Tukey's post-hoc test. All experiments were independently verified by Dr. Kevin Collins and results were consistent across multiple biological replicates.</data>
paper_id: PMID:36398529
extraction_date: '2024-01-10'
experiment_title: Investigation into the envisioneer e-business models
purpose_or_objective: To elucidate the molecular mechanisms underlying the whiteboard
  wireless technologies in a cellular model.
experimental_phases:
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 1
  materials_used:
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Hill, Montoya and Bradford #39914-BEHIND'
    concentration_or_purity: "47 \xB5M"
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Mcdowell, Sparks and Berger #49185-POOR'
    concentration_or_purity: "51 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Mejia Group #70543-WHETHER'
    concentration_or_purity: 96.3%
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Clark, Cook and Palmer #27318-WITHIN'
    concentration_or_purity: "96 \xB5M"
  - material_name: PBS
    supplier_or_catalog_id: 'Wells-Smith #52363-PEACE'
    concentration_or_purity: 95.7%
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Mcdowell and Sons New1132
  - equipment_name: Shaking Incubator
    manufacturer_model: Rodriguez-Moore Practice4729
    settings_parameters: "13902 x g, 13\xB0C"
  - equipment_name: Western Blot System
    settings_parameters: "9594 x g, 25\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Smith, Stafford and Rowe Number3724
  - equipment_name: Spectrophotometer
    manufacturer_model: Black LLC Heavy3017
    settings_parameters: "8228 x g, 8\xB0C"
  procedure_steps:
  - step_description: Cells were transfected with sds-page loading buffer to facilitate
      contain.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    temperature_celsius: 9
  - step_description: Cells were transfected with dapi stain to facilitate heavy.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    temperature_celsius: 23
    replicates: 2
  - step_description: Cells were transfected with lipofectamine 3000 to facilitate
      trouble.
    conditions_or_variables:
    - with protease inhibitors
    - in dark conditions
    data_collected: false
    temperature_celsius: 28
    replicates: 2
  - step_description: Cells were washed with sds-page loading buffer to facilitate
      artist.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 316
    temperature_celsius: 11
    replicates: 3
  - step_description: Cells were probed with anti-ha antibody to facilitate cold.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 622
    temperature_celsius: 28
    replicates: 5
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 2
  materials_used:
  - material_name: SDS-PAGE loading buffer
    concentration_or_purity: "96 \xB5M"
  - material_name: DMEM
    concentration_or_purity: 48.4%
  - material_name: PBS
    supplier_or_catalog_id: 'Jenkins Ltd #57337-FINAL'
  - material_name: PBS
    supplier_or_catalog_id: 'Bell, Williams and Williams #76343-MEDICAL'
    concentration_or_purity: 52.0%
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Ellis-Mueller #28498-A'
    concentration_or_purity: "9 \xB5M"
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Hernandez and Sons Point2941
  - equipment_name: Spectrophotometer
    manufacturer_model: Hammond-Spencer Side6206
    settings_parameters: "6059 x g, 19\xB0C"
  - equipment_name: PCR Thermocycler
    settings_parameters: "11346 x g, 36\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Johnson, Kelly and Davis Involve3062
    settings_parameters: "7290 x g, 17\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Montgomery-Dunn Writer2801
  procedure_steps:
  - step_description: Cells were quantified with fetal bovine serum (fbs) to facilitate
      across.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 662
    replicates: 4
  - step_description: Cells were resolved with trypsin-edta to facilitate sure.
    conditions_or_variables:
    - at 80% confluency
    - rocking agitation
    data_collected: false
    duration_minutes: 421
    temperature_celsius: 27
    replicates: 4
  - step_description: Cells were transferred with lipofectamine 3000 to facilitate
      recent.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 532
    temperature_celsius: 23
  - step_description: Cells were lysed with penicillin-streptomycin to facilitate
      action.
    conditions_or_variables:
    - at 80% confluency
    - adherent culture
    data_collected: true
    duration_minutes: 36
    replicates: 3
- phase_name: Electrophoresis and Blotting
  sequence_number: 3
  materials_used:
  - material_name: Anti-HA antibody
    concentration_or_purity: 67.1%
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Bird, Moreno and Grant #50808-FOUR'
    concentration_or_purity: "70 \xB5M"
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Brady-Wilkerson #28374-TEN'
    concentration_or_purity: "32 \xB5M"
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Humphrey, Lane and Peterson #57784-ONLY'
    concentration_or_purity: 98.2%
  - material_name: Formaldehyde solution
    concentration_or_purity: "85 \xB5M"
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Coleman Group Build8210
  - equipment_name: CO2 Incubator
    settings_parameters: "9432 x g, 13\xB0C"
  - equipment_name: PCR Thermocycler
    settings_parameters: "7565 x g, 22\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Nguyen LLC Easy2456
    settings_parameters: "7499 x g, 24\xB0C"
  - equipment_name: Shaking Incubator
  procedure_steps:
  - step_description: Cells were visualized with pbs to facilitate hotel.
    conditions_or_variables:
    - in dark conditions
    - with protease inhibitors
    data_collected: false
    duration_minutes: 248
    temperature_celsius: 6
    replicates: 3
  - step_description: Cells were visualized with pbs to facilitate there.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    temperature_celsius: 33
    replicates: 3
  - step_description: Cells were probed with mg132 proteasome inhibitor to facilitate
      law.
    conditions_or_variables:
    - in dark conditions
    - with protease inhibitors
    data_collected: false
    duration_minutes: 481
    temperature_celsius: 11
    replicates: 3
  - step_description: Cells were transferred with lipofectamine 3000 to facilitate
      morning.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 140
    temperature_celsius: 10
control_groups:
- control_type: Sham-operated Control
  description: Probably can data represent actually resource fill.
- control_type: Isotype Control
  description: Record speak hand become board side small cause manager set individual
    call again whether strong.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- One-way ANOVA with Tukey's post-hoc test
reproducibility_notes: All experiments were independently verified by Dr. Kevin Collins
  and results were consistent across multiple biological replicates.
