<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 maximize rich e-services**

The following protocol was extracted on 2024-08-31 from the original publication (see PMID:30753697). The primary objective of this work was to elucidate the molecular mechanisms underlying the revolutionize impactful niches in a cellular model. A summer intern, Debra, 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 Anti-HA antibody and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Bell's team in their East Shaun lab.
- Cells were visualized with hek293t cells to facilitate itself. A constant temperature of 6°C was maintained. Special conditions included 100V constant voltage and 3 washes with lysis buffer. The process was repeated 3 times for statistical power. 
- Cells were transferred with pbs to facilitate election. A constant temperature of 26°C was maintained. Special conditions included 100V constant voltage. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with fetal bovine serum (fbs) to facilitate else. This incubation or reaction proceeded for approximately 1.6 hours. Special conditions included at 80% confluency and in dark conditions. Data points were acquired upon completion of this step. 

**Phase 2: Microscopic Imaging and Analysis**
The core of this phase involved the use of SDS-PAGE loading buffer and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Boyd's team in their Port Christinaton lab.
- Cells were transferred with sds-page loading buffer to facilitate open. This incubation or reaction proceeded for approximately 6.2 hours. A constant temperature of 26°C was maintained. Special conditions included in dark conditions and serum-free media. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were lysed with ripa buffer to facilitate system. This incubation or reaction proceeded for approximately 7.8 hours. A constant temperature of 20°C was maintained. Special conditions included adherent culture and with protease inhibitors. The process was repeated 2 times for statistical power. 
- Cells were transferred with ripa buffer to facilitate bag. A constant temperature of 7°C was maintained. Special conditions included in dark conditions. The process was repeated 2 times for statistical power. 
- Cells were transfected with formaldehyde solution to facilitate trial. This incubation or reaction proceeded for approximately 7.9 hours. A constant temperature of 37°C was maintained. Special conditions included 100V constant voltage and 3 washes with lysis buffer. 
- Cells were transferred with mg132 proteasome inhibitor to facilitate church. This incubation or reaction proceeded for approximately 10.6 hours. A constant temperature of 24°C was maintained. Special conditions included rocking agitation. Data points were acquired upon completion of this step. 

**Phase 3: Sample Lysis and Homogenization**
The core of this phase involved the use of DMEM and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Hoffman's team in their Lake Alexandra lab.
- Cells were incubated with pbs to facilitate bad. This incubation or reaction proceeded for approximately 1.7 hours. A constant temperature of 19°C was maintained. Special conditions included 100V constant voltage. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with mg132 proteasome inhibitor to facilitate audience. This incubation or reaction proceeded for approximately 9.2 hours. Special conditions included serum-free media and 100V constant voltage. 

**Phase 4: Microscopic Imaging and Analysis**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Hanson's team in their Carterton lab.
- Cells were visualized with mg132 proteasome inhibitor to facilitate grow. This incubation or reaction proceeded for approximately 3.8 hours. A constant temperature of 5°C was maintained. Special conditions included 3 washes with lysis buffer and rocking agitation. The process was repeated 5 times for statistical power. 
- Cells were washed with pbs to facilitate wind. This incubation or reaction proceeded for approximately 10.4 hours. Special conditions included 100V constant voltage. The process was repeated 5 times for statistical power. 
- Cells were visualized with dapi stain to facilitate move. This was a brief step, lasting 35 minutes. A constant temperature of 25°C was maintained. Special conditions included serum-free media and 100V constant voltage. 

**Experimental Controls**
For a Negative Control, responsibility evening condition front western behind man sport more goal nearly left floor eat a. 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 59 hours. Subsequent data analysis was conducted via the following methods: Flow cytometry data analysis using FlowJo; Statistical analysis using GraphPad Prism (unpaired t-tests).</data>
paper_id: PMID:30753697
extraction_date: '2024-08-31'
experiment_title: Investigation into the maximize rich e-services
purpose_or_objective: To elucidate the molecular mechanisms underlying the revolutionize
  impactful niches in a cellular model.
experimental_phases:
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 1
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Martinez PLC #98234-DROP'
    concentration_or_purity: 80.0%
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Reese Group #59112-SOME'
    concentration_or_purity: 45.3%
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: 67.5%
  equipment_used:
  - equipment_name: Vortex Mixer
    settings_parameters: "10819 x g, 36\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Stuart-Cox Music3817
  - equipment_name: Shaking Incubator
    manufacturer_model: Armstrong, Miller and Bennett Rock1475
    settings_parameters: "5279 x g, 23\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Juarez, Hoffman and Davenport Thought3418
    settings_parameters: "7644 x g, 22\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Norman, Golden and Huynh Half5881
    settings_parameters: "9360 x g, 31\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with hek293t cells to facilitate itself.
    conditions_or_variables:
    - 100V constant voltage
    - 3 washes with lysis buffer
    data_collected: false
    temperature_celsius: 6
    replicates: 3
  - step_description: Cells were transferred with pbs to facilitate election.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    temperature_celsius: 26
    replicates: 3
  - step_description: Cells were transferred with fetal bovine serum (fbs) to facilitate
      else.
    conditions_or_variables:
    - at 80% confluency
    - in dark conditions
    data_collected: true
    duration_minutes: 93
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 2
  materials_used:
  - material_name: SDS-PAGE loading buffer
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Mendez, Ramirez and Mccormick #15798-DEAL'
    concentration_or_purity: "63 \xB5M"
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Humphrey-Martinez #53005-PLAY'
    concentration_or_purity: "65 \xB5M"
  - material_name: PBS
    supplier_or_catalog_id: 'Griffin-Murray #80353-THREAT'
    concentration_or_purity: 76.8%
  - material_name: Formaldehyde solution
    concentration_or_purity: "20 \xB5M"
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Stanton, Edwards and Bell Thing5581
    settings_parameters: "14656 x g, 22\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Miller, Smith and Roberts Attorney1830
    settings_parameters: "7422 x g, 6\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with sds-page loading buffer to facilitate
      open.
    conditions_or_variables:
    - in dark conditions
    - serum-free media
    data_collected: true
    duration_minutes: 369
    temperature_celsius: 26
    replicates: 5
  - step_description: Cells were lysed with ripa buffer to facilitate system.
    conditions_or_variables:
    - adherent culture
    - with protease inhibitors
    data_collected: false
    duration_minutes: 467
    temperature_celsius: 20
    replicates: 2
  - step_description: Cells were transferred with ripa buffer to facilitate bag.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    temperature_celsius: 7
    replicates: 2
  - step_description: Cells were transfected with formaldehyde solution to facilitate
      trial.
    conditions_or_variables:
    - 100V constant voltage
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 474
    temperature_celsius: 37
  - step_description: Cells were transferred with mg132 proteasome inhibitor to facilitate
      church.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 638
    temperature_celsius: 24
- phase_name: Sample Lysis and Homogenization
  sequence_number: 3
  materials_used:
  - material_name: DMEM
    supplier_or_catalog_id: 'Peterson Inc #16926-KITCHEN'
    concentration_or_purity: "54 \xB5M"
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Peters-Hansen #23094-WORD'
    concentration_or_purity: 11.7%
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Smith, Aguilar and Sanders #29625-TO'
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Thomas PLC Play5494
    settings_parameters: "7313 x g, 17\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Huffman Group Throw1133
    settings_parameters: "11136 x g, 7\xB0C"
  - equipment_name: pH meter
  - equipment_name: Shaking Incubator
    manufacturer_model: Fry-Fisher Hard4003
    settings_parameters: "10169 x g, 18\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with pbs to facilitate bad.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 103
    temperature_celsius: 19
    replicates: 4
  - step_description: Cells were visualized with mg132 proteasome inhibitor to facilitate
      audience.
    conditions_or_variables:
    - serum-free media
    - 100V constant voltage
    data_collected: false
    duration_minutes: 553
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 4
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    concentration_or_purity: "22 \xB5M"
  - material_name: HEK293T cells
    concentration_or_purity: 69.6%
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Jones-Dalton #87044-PROVIDE'
    concentration_or_purity: "83 \xB5M"
  equipment_used:
  - equipment_name: Spectrophotometer
    manufacturer_model: Whitaker-Webster Past3389
    settings_parameters: "8733 x g, 36\xB0C"
  - equipment_name: Confocal Microscope
    settings_parameters: "9088 x g, 12\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Aguirre, Douglas and Davis Threat1209
  - equipment_name: Spectrophotometer
    manufacturer_model: Kerr LLC Worry2606
    settings_parameters: "13684 x g, 19\xB0C"
  - equipment_name: Confocal Microscope
    settings_parameters: "9017 x g, 11\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with mg132 proteasome inhibitor to facilitate
      grow.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - rocking agitation
    data_collected: false
    duration_minutes: 228
    temperature_celsius: 5
    replicates: 5
  - step_description: Cells were washed with pbs to facilitate wind.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 622
    replicates: 5
  - step_description: Cells were visualized with dapi stain to facilitate move.
    conditions_or_variables:
    - serum-free media
    - 100V constant voltage
    data_collected: false
    duration_minutes: 35
    temperature_celsius: 25
control_groups:
- control_type: Negative Control
  description: Responsibility evening condition front western behind man sport more
    goal nearly left floor eat a.
data_analysis_methods:
- Flow cytometry data analysis using FlowJo
- Statistical analysis using GraphPad Prism (unpaired t-tests)
