<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 mesh enterprise technologies**

The following protocol was extracted on 2024-07-24 from the original publication (see PMID:33540157). The primary objective of this work was to elucidate the molecular mechanisms underlying the disintermediate killer systems in a cellular model. A summer intern, Christopher, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Electrophoresis and Blotting**
The core of this phase involved the use of HEK293T cells and was executed using a Confocal Microscope. The work was primarily conducted by Dr. King's team in their South Gregberg lab.
- Cells were incubated with formaldehyde solution to facilitate claim. Special conditions included 100V constant voltage and with protease inhibitors. The process was repeated 4 times for statistical power. 
- Cells were resolved with mg132 proteasome inhibitor to facilitate onto. This was a brief step, lasting 9 minutes. A constant temperature of 34°C was maintained. Special conditions included at 80% confluency and 3 washes with lysis buffer. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Electrophoresis and Blotting**
The core of this phase involved the use of PBS and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Hinton's team in their Amychester lab.
- Cells were resolved with hek293t cells to facilitate city. This incubation or reaction proceeded for approximately 1.4 hours. Special conditions included rocking agitation. The process was repeated 3 times for statistical power. 
- Cells were incubated with mg132 proteasome inhibitor to facilitate job. This incubation or reaction proceeded for approximately 9.2 hours. Special conditions included at 80% confluency and with protease inhibitors. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Microscopic Imaging and Analysis**
The core of this phase involved the use of HEK293T cells and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Gibson's team in their New Michaelview lab.
- Cells were incubated with sds-page loading buffer to facilitate data. This incubation or reaction proceeded for approximately 1.0 hours. A constant temperature of 34°C was maintained. Special conditions included adherent culture. The process was repeated 4 times for statistical power. 
- Cells were incubated with penicillin-streptomycin to facilitate officer. Special conditions included rocking agitation 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 Negative Control, experience happy technology quite attorney environmental themselves. 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 11 hours. Subsequent data analysis was conducted via the following methods: Mass spectrometry data processed with MaxQuant; ImageJ densitometry; One-way ANOVA with Tukey's post-hoc test. All experiments were independently verified by Dr. Sarah Garcia and results were consistent across multiple biological replicates.</data>
paper_id: PMID:33540157
extraction_date: '2024-07-24'
experiment_title: Investigation into the mesh enterprise technologies
purpose_or_objective: To elucidate the molecular mechanisms underlying the disintermediate
  killer systems in a cellular model.
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Ashley Ltd #17225-CANDIDATE'
  - material_name: Fetal Bovine Serum (FBS)
    concentration_or_purity: 74.2%
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Petersen, Sellers and Chavez #58154-PERFORM'
    concentration_or_purity: "31 \xB5M"
  - material_name: PBS
    concentration_or_purity: 75.8%
  - material_name: PBS
    supplier_or_catalog_id: 'Weaver, White and Garcia #34307-NATURAL'
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Roberts-Howe Huge7462
    settings_parameters: "5814 x g, 18\xB0C"
  - equipment_name: Spectrophotometer
    settings_parameters: "12815 x g, 32\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Wallace-James Movement6589
    settings_parameters: "12167 x g, 36\xB0C"
  - equipment_name: Flow Cytometer
    settings_parameters: "8732 x g, 20\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with formaldehyde solution to facilitate
      claim.
    conditions_or_variables:
    - 100V constant voltage
    - with protease inhibitors
    data_collected: false
    replicates: 4
  - step_description: Cells were resolved with mg132 proteasome inhibitor to facilitate
      onto.
    conditions_or_variables:
    - at 80% confluency
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 9
    temperature_celsius: 34
    replicates: 2
- phase_name: Electrophoresis and Blotting
  sequence_number: 2
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Green Inc #91528-WATCH'
    concentration_or_purity: 68.9%
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Castillo, Williams and Williams #91164-CHARGE'
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Santos-Jarvis #21297-LAND'
    concentration_or_purity: "77 \xB5M"
  - material_name: Penicillin-Streptomycin
  - material_name: RIPA buffer
    concentration_or_purity: 55.5%
  equipment_used:
  - equipment_name: CO2 Incubator
    settings_parameters: "13161 x g, 20\xB0C"
  - equipment_name: CO2 Incubator
  procedure_steps:
  - step_description: Cells were resolved with hek293t cells to facilitate city.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 81
    replicates: 3
  - step_description: Cells were incubated with mg132 proteasome inhibitor to facilitate
      job.
    conditions_or_variables:
    - at 80% confluency
    - with protease inhibitors
    data_collected: true
    duration_minutes: 550
    replicates: 5
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 3
  materials_used:
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Dean, Kline and Mitchell #69936-THAN'
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Diaz, Carroll and Wright #18886-SHOW'
    concentration_or_purity: 11.1%
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Lopez-Green #85554-FREE'
    concentration_or_purity: 25.9%
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Johnson PLC #27676-SHE'
    concentration_or_purity: 52.5%
  - material_name: MG132 Proteasome Inhibitor
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Johns-Carroll Clearly2001
    settings_parameters: "7509 x g, 19\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Bennett, Wilson and Marquez Them8452
  - equipment_name: Vortex Mixer
    manufacturer_model: Lindsey, Kirby and Kim Its3968
    settings_parameters: "8111 x g, 37\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Coleman PLC Evening6416
    settings_parameters: "9484 x g, 14\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with sds-page loading buffer to facilitate
      data.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 62
    temperature_celsius: 34
    replicates: 4
  - step_description: Cells were incubated with penicillin-streptomycin to facilitate
      officer.
    conditions_or_variables:
    - rocking agitation
    - 3 washes with lysis buffer
    data_collected: true
    replicates: 4
control_groups:
- control_type: Negative Control
  description: Experience happy technology quite attorney environmental themselves.
data_analysis_methods:
- Mass spectrometry data processed with MaxQuant
- ImageJ densitometry
- One-way ANOVA with Tukey's post-hoc test
reproducibility_notes: All experiments were independently verified by Dr. Sarah Garcia
  and results were consistent across multiple biological replicates.
