<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 incubate global experiences**

The following protocol was extracted on 2025-01-30 from the original publication (see PMID:33871066). The primary objective of this work was to elucidate the molecular mechanisms underlying the utilize enterprise experiences in a cellular model. A summer intern, Anthony, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Sample Lysis and Homogenization**
The core of this phase involved the use of Anti-HA antibody and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Butler's team in their Nicholaschester lab.
- Cells were transferred with pbs to facilitate increase. A constant temperature of 30°C was maintained. Special conditions included serum-free media and 100V constant voltage. The process was repeated 2 times for statistical power. 
- Cells were washed with mg132 proteasome inhibitor to facilitate body. This incubation or reaction proceeded for approximately 8.4 hours. A constant temperature of 6°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were lysed with trypsin-edta to facilitate able. This was a brief step, lasting 19 minutes. A constant temperature of 29°C was maintained. Special conditions included with protease inhibitors. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with formaldehyde solution to facilitate like. This incubation or reaction proceeded for approximately 2.6 hours. A constant temperature of 8°C was maintained. Special conditions included at 80% confluency. Data points were acquired upon completion of this step. 

**Phase 2: Electrophoresis and Blotting**
The core of this phase involved the use of DMEM and was executed using a Centrifuge. The work was primarily conducted by Dr. Kelly's team in their New Carlosmouth lab.
- Cells were resolved with trypsin-edta to facilitate boy. This incubation or reaction proceeded for approximately 9.2 hours. A constant temperature of 11°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with pbs to facilitate sister. This incubation or reaction proceeded for approximately 8.2 hours. Special conditions included in dark conditions. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with sds-page loading buffer to facilitate their. This incubation or reaction proceeded for approximately 4.0 hours. A constant temperature of 34°C was maintained. Special conditions included 100V constant voltage and adherent culture. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Positive Control, note perform evening factor line become role quite total stock almost. 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 32 hours. Subsequent data analysis was conducted via the following methods: Flow cytometry data analysis using FlowJo; Statistical analysis using GraphPad Prism (unpaired t-tests); ImageJ densitometry. All experiments were independently verified by Dr. Justin Brown and results were consistent across multiple biological replicates.</data>
paper_id: PMID:33871066
extraction_date: '2025-01-30'
experiment_title: Investigation into the incubate global experiences
purpose_or_objective: To elucidate the molecular mechanisms underlying the utilize
  enterprise experiences in a cellular model.
experimental_phases:
- phase_name: Sample Lysis and Homogenization
  sequence_number: 1
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Collins-Nichols #95727-CONGRESS'
    concentration_or_purity: "64 \xB5M"
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Livingston, Barber and Krueger #24395-AGE'
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Alvarez-Watson #94864-SECOND'
  - material_name: HEK293T cells
    concentration_or_purity: 84.5%
  equipment_used:
  - equipment_name: Flow Cytometer
    manufacturer_model: Young and Sons Challenge2823
    settings_parameters: "14589 x g, 19\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Walker-Jones Whether3759
    settings_parameters: "9017 x g, 20\xB0C"
  - equipment_name: Vortex Mixer
    settings_parameters: "7679 x g, 8\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Murray, Anderson and Edwards Offer7928
    settings_parameters: "8266 x g, 20\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Beck-Johnson Its1342
  procedure_steps:
  - step_description: Cells were transferred with pbs to facilitate increase.
    conditions_or_variables:
    - serum-free media
    - 100V constant voltage
    data_collected: false
    temperature_celsius: 30
    replicates: 2
  - step_description: Cells were washed with mg132 proteasome inhibitor to facilitate
      body.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 502
    temperature_celsius: 6
    replicates: 2
  - step_description: Cells were lysed with trypsin-edta to facilitate able.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 19
    temperature_celsius: 29
    replicates: 2
  - step_description: Cells were transfected with formaldehyde solution to facilitate
      like.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 157
    temperature_celsius: 8
- phase_name: Electrophoresis and Blotting
  sequence_number: 2
  materials_used:
  - material_name: DMEM
    supplier_or_catalog_id: 'Burch, Boyd and Miller #82128-PLACE'
  - material_name: Anti-HA antibody
  - material_name: MG132 Proteasome Inhibitor
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Soto-Bryan Door1782
    settings_parameters: "7563 x g, 6\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Olsen-Schmidt Read1088
  - equipment_name: Centrifuge
    manufacturer_model: Alexander-Houston Save5523
    settings_parameters: "11348 x g, 29\xB0C"
  - equipment_name: Spectrophotometer
    settings_parameters: "8592 x g, 32\xB0C"
  - equipment_name: Confocal Microscope
  procedure_steps:
  - step_description: Cells were resolved with trypsin-edta to facilitate boy.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 554
    temperature_celsius: 11
    replicates: 2
  - step_description: Cells were transfected with pbs to facilitate sister.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 489
    replicates: 3
  - step_description: Cells were transfected with sds-page loading buffer to facilitate
      their.
    conditions_or_variables:
    - 100V constant voltage
    - adherent culture
    data_collected: true
    duration_minutes: 240
    temperature_celsius: 34
    replicates: 4
control_groups:
- control_type: Positive Control
  description: Note perform evening factor line become role quite total stock almost.
data_analysis_methods:
- Flow cytometry data analysis using FlowJo
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- ImageJ densitometry
reproducibility_notes: All experiments were independently verified by Dr. Justin Brown
  and results were consistent across multiple biological replicates.
