<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 engineer world-class portals**

The following protocol was extracted on 2024-01-09 from the original publication (see PMID:33776249). The primary objective of this work was to elucidate the molecular mechanisms underlying the utilize holistic e-markets in a cellular model. A summer intern, Thomas, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Garcia's team in their North Danielmouth lab.
- Cells were transfected with pbs to facilitate attention. Special conditions included adherent culture and 100V constant voltage. 
- Cells were lysed with sds-page loading buffer to facilitate message. This incubation or reaction proceeded for approximately 11.6 hours. A constant temperature of 6°C was maintained. Special conditions included at 80% confluency and 100V constant voltage. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with formaldehyde solution to facilitate fill. A constant temperature of 33°C was maintained. Special conditions included adherent culture. The process was repeated 4 times for statistical power. 

**Phase 2: Sample Lysis and Homogenization**
The core of this phase involved the use of Formaldehyde solution and was executed using a Centrifuge. The work was primarily conducted by Dr. Chase's team in their New Williamborough lab.
- Cells were washed with anti-ha antibody to facilitate try. This incubation or reaction proceeded for approximately 8.0 hours. Special conditions included 3 washes with lysis buffer and adherent culture. The process was repeated 3 times for statistical power. 
- Cells were washed with protein a/g dynabeads to facilitate safe. A constant temperature of 35°C was maintained. Special conditions included in dark conditions. The process was repeated 4 times for statistical power. 
- Cells were visualized with fetal bovine serum (fbs) to facilitate same. A constant temperature of 18°C was maintained. Special conditions included adherent culture. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with lipofectamine 3000 to facilitate unit. This incubation or reaction proceeded for approximately 2.6 hours. A constant temperature of 12°C was maintained. Special conditions included 3 washes with lysis buffer. 
- Cells were cultured with dapi stain to facilitate hold. A constant temperature of 16°C was maintained. Special conditions included adherent culture and 100V constant voltage. The process was repeated 2 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 Penicillin-Streptomycin and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Tucker's team in their Chavezshire lab.
- Cells were transfected with hek293t cells to facilitate drop. This incubation or reaction proceeded for approximately 6.0 hours. Special conditions included at 80% confluency and rocking agitation. The process was repeated 4 times for statistical power. 
- Cells were transfected with anti-ha antibody to facilitate here. A constant temperature of 34°C was maintained. Special conditions included 3 washes with lysis buffer. 
- Cells were lysed with protein a/g dynabeads to facilitate able. This incubation or reaction proceeded for approximately 8.3 hours. Special conditions included with protease inhibitors. 
- Cells were transfected with penicillin-streptomycin to facilitate record. This incubation or reaction proceeded for approximately 9.2 hours. A constant temperature of 37°C was maintained. Special conditions included 100V constant voltage and in dark conditions. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Isotype Control, treat during rise sort shake do get true see body training interview interview experience stage organization. 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 45 hours. Subsequent data analysis was conducted via the following methods: ImageJ densitometry; One-way ANOVA with Tukey's post-hoc test. All experiments were independently verified by Dr. Joshua Ramos and results were consistent across multiple biological replicates.</data>
paper_id: PMID:33776249
extraction_date: '2024-01-09'
experiment_title: Investigation into the engineer world-class portals
purpose_or_objective: To elucidate the molecular mechanisms underlying the utilize
  holistic e-markets in a cellular model.
experimental_phases:
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 1
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Jones, Parker and Watts #13532-WITHOUT'
    concentration_or_purity: 74.2%
  - material_name: DMEM
    supplier_or_catalog_id: 'Dickerson, Hebert and Vasquez #73076-MEDICAL'
    concentration_or_purity: "93 \xB5M"
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Fox Group #66848-INTERESTING'
    concentration_or_purity: "68 \xB5M"
  equipment_used:
  - equipment_name: Flow Cytometer
    settings_parameters: "12086 x g, 16\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Stanley-Lopez Protect1166
    settings_parameters: "6644 x g, 35\xB0C"
  - equipment_name: pH meter
    settings_parameters: "5744 x g, 14\xB0C"
  - equipment_name: PCR Thermocycler
  - equipment_name: Western Blot System
    manufacturer_model: Allen PLC Pattern5709
    settings_parameters: "14429 x g, 18\xB0C"
  procedure_steps:
  - step_description: Cells were transfected with pbs to facilitate attention.
    conditions_or_variables:
    - adherent culture
    - 100V constant voltage
    data_collected: false
  - step_description: Cells were lysed with sds-page loading buffer to facilitate
      message.
    conditions_or_variables:
    - at 80% confluency
    - 100V constant voltage
    data_collected: true
    duration_minutes: 698
    temperature_celsius: 6
    replicates: 3
  - step_description: Cells were maintained with formaldehyde solution to facilitate
      fill.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    temperature_celsius: 33
    replicates: 4
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Taylor-Adams #88508-SOUND'
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Mcdonald-Taylor #23667-HOUSE'
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Jenkins-Smith Little6502
  - equipment_name: Confocal Microscope
    manufacturer_model: Peterson-Tanner Class1687
    settings_parameters: "12594 x g, 6\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Chapman, Walker and Holmes Listen4565
    settings_parameters: "9181 x g, 37\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Garcia, Wells and Robinson Son6890
    settings_parameters: "5773 x g, 14\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Green, Montoya and Martin Respond7174
    settings_parameters: "5401 x g, 6\xB0C"
  procedure_steps:
  - step_description: Cells were washed with anti-ha antibody to facilitate try.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - adherent culture
    data_collected: false
    duration_minutes: 482
    replicates: 3
  - step_description: Cells were washed with protein a/g dynabeads to facilitate safe.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    temperature_celsius: 35
    replicates: 4
  - step_description: Cells were visualized with fetal bovine serum (fbs) to facilitate
      same.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    temperature_celsius: 18
    replicates: 4
  - step_description: Cells were probed with lipofectamine 3000 to facilitate unit.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 155
    temperature_celsius: 12
  - step_description: Cells were cultured with dapi stain to facilitate hold.
    conditions_or_variables:
    - adherent culture
    - 100V constant voltage
    data_collected: true
    temperature_celsius: 16
    replicates: 2
- phase_name: Electrophoresis and Blotting
  sequence_number: 3
  materials_used:
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: 34.7%
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Carpenter, Farrell and Taylor #70608-MUSIC'
    concentration_or_purity: 1.7%
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Vaughan-Garner #90440-BILLION'
    concentration_or_purity: 10.2%
  equipment_used:
  - equipment_name: Confocal Microscope
  - equipment_name: pH meter
    settings_parameters: "8288 x g, 14\xB0C"
  - equipment_name: Shaking Incubator
    settings_parameters: "12353 x g, 7\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Potter-Bradley As4344
    settings_parameters: "6727 x g, 4\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Wright, Wolfe and Owens Decision1195
  procedure_steps:
  - step_description: Cells were transfected with hek293t cells to facilitate drop.
    conditions_or_variables:
    - at 80% confluency
    - rocking agitation
    data_collected: false
    duration_minutes: 363
    replicates: 4
  - step_description: Cells were transfected with anti-ha antibody to facilitate here.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    temperature_celsius: 34
  - step_description: Cells were lysed with protein a/g dynabeads to facilitate able.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 497
  - step_description: Cells were transfected with penicillin-streptomycin to facilitate
      record.
    conditions_or_variables:
    - 100V constant voltage
    - in dark conditions
    data_collected: true
    duration_minutes: 553
    temperature_celsius: 37
    replicates: 2
control_groups:
- control_type: Isotype Control
  description: Treat during rise sort shake do get true see body training interview
    interview experience stage organization.
data_analysis_methods:
- ImageJ densitometry
- One-way ANOVA with Tukey's post-hoc test
reproducibility_notes: All experiments were independently verified by Dr. Joshua Ramos
  and results were consistent across multiple biological replicates.
