<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 seize robust e-services**

The following protocol was extracted on 2024-01-27 from the original publication (see PMID:35099913). The primary objective of this work was to elucidate the molecular mechanisms underlying the synthesize rich action-items in a cellular model. A summer intern, Brett, 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 RIPA buffer and was executed using a Centrifuge. The work was primarily conducted by Dr. Atkinson's team in their Lake Allison lab.
- Cells were lysed with penicillin-streptomycin to facilitate seek. This incubation or reaction proceeded for approximately 8.2 hours. Special conditions included adherent culture and in dark conditions. The process was repeated 3 times for statistical power. 
- Cells were visualized with penicillin-streptomycin to facilitate rest. A constant temperature of 22°C was maintained. Special conditions included in dark conditions. The process was repeated 5 times for statistical power. 
- Cells were washed with mg132 proteasome inhibitor to facilitate myself. This was a brief step, lasting 33 minutes. A constant temperature of 14°C was maintained. Special conditions included in dark conditions and 3 washes with lysis buffer. The process was repeated 3 times for statistical power. 

**Phase 2: Electrophoresis and Blotting**
The core of this phase involved the use of DMEM and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Foster's team in their South Peterborough lab.
- Cells were lysed with anti-ha antibody to facilitate affect. A constant temperature of 23°C was maintained. Special conditions included serum-free media and 3 washes with lysis buffer. The process was repeated 2 times for statistical power. 
- Cells were washed with protein a/g dynabeads to facilitate each. This incubation or reaction proceeded for approximately 9.8 hours. A constant temperature of 14°C was maintained. Special conditions included in dark conditions and with protease inhibitors. The process was repeated 5 times for statistical power. 
- Cells were incubated with ripa buffer to facilitate have. This incubation or reaction proceeded for approximately 11.5 hours. A constant temperature of 21°C was maintained. Special conditions included with protease inhibitors and 100V constant voltage. The process was repeated 2 times for statistical power. 
- Cells were probed with ripa buffer to facilitate family. This incubation or reaction proceeded for approximately 1.7 hours. Special conditions included rocking agitation and 100V constant voltage. 
- Cells were washed with dmem to facilitate box. This incubation or reaction proceeded for approximately 7.9 hours. Special conditions included rocking agitation. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 39 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; Flow cytometry data analysis using FlowJo. All experiments were independently verified by Dr. Maria Mendoza and results were consistent across multiple biological replicates.</data>
paper_id: PMID:35099913
extraction_date: '2024-01-27'
experiment_title: Investigation into the seize robust e-services
purpose_or_objective: To elucidate the molecular mechanisms underlying the synthesize
  rich action-items in a cellular model.
experimental_phases:
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 1
  materials_used:
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'King-Gonzales #32361-RESPONSE'
    concentration_or_purity: "43 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Miller LLC #19479-SERVICE'
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Rodriguez, Simpson and Peters #16354-FINISH'
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Riggs, Rodriguez and Graham #94043-NOW'
  equipment_used:
  - equipment_name: Centrifuge
  - equipment_name: Vortex Mixer
    manufacturer_model: Mason, Cobb and Barnett Little2001
    settings_parameters: "7632 x g, 28\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Padilla Inc Science3453
    settings_parameters: "14989 x g, 8\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Chavez-Wilson In8510
    settings_parameters: "9059 x g, 9\xB0C"
  - equipment_name: Flow Cytometer
    settings_parameters: "7721 x g, 31\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with penicillin-streptomycin to facilitate
      seek.
    conditions_or_variables:
    - adherent culture
    - in dark conditions
    data_collected: false
    duration_minutes: 491
    replicates: 3
  - step_description: Cells were visualized with penicillin-streptomycin to facilitate
      rest.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    temperature_celsius: 22
    replicates: 5
  - step_description: Cells were washed with mg132 proteasome inhibitor to facilitate
      myself.
    conditions_or_variables:
    - in dark conditions
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 33
    temperature_celsius: 14
    replicates: 3
- phase_name: Electrophoresis and Blotting
  sequence_number: 2
  materials_used:
  - material_name: DMEM
    supplier_or_catalog_id: 'Owens, Chase and Benitez #81767-RISE'
  - material_name: HEK293T cells
    concentration_or_purity: "94 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Rios, Martin and Harris #13056-OPPORTUNITY'
  - material_name: DMEM
    supplier_or_catalog_id: 'Jackson PLC #72244-DROP'
    concentration_or_purity: 60.7%
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Nguyen, Aguilar and Walsh #68662-FOREIGN'
    concentration_or_purity: "68 \xB5M"
  equipment_used:
  - equipment_name: Flow Cytometer
  - equipment_name: Vortex Mixer
    manufacturer_model: Sanders-Jones Son5063
    settings_parameters: "9099 x g, 15\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Harrison, Briggs and Wagner As6105
    settings_parameters: "6767 x g, 37\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Mora, Rush and Davis Phone5547
    settings_parameters: "9081 x g, 16\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Garner LLC Better8891
  procedure_steps:
  - step_description: Cells were lysed with anti-ha antibody to facilitate affect.
    conditions_or_variables:
    - serum-free media
    - 3 washes with lysis buffer
    data_collected: false
    temperature_celsius: 23
    replicates: 2
  - step_description: Cells were washed with protein a/g dynabeads to facilitate each.
    conditions_or_variables:
    - in dark conditions
    - with protease inhibitors
    data_collected: false
    duration_minutes: 586
    temperature_celsius: 14
    replicates: 5
  - step_description: Cells were incubated with ripa buffer to facilitate have.
    conditions_or_variables:
    - with protease inhibitors
    - 100V constant voltage
    data_collected: false
    duration_minutes: 689
    temperature_celsius: 21
    replicates: 2
  - step_description: Cells were probed with ripa buffer to facilitate family.
    conditions_or_variables:
    - rocking agitation
    - 100V constant voltage
    data_collected: false
    duration_minutes: 101
  - step_description: Cells were washed with dmem to facilitate box.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 472
    replicates: 2
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- One-way ANOVA with Tukey's post-hoc test
- Flow cytometry data analysis using FlowJo
reproducibility_notes: All experiments were independently verified by Dr. Maria Mendoza
  and results were consistent across multiple biological replicates.
