<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 facilitate turn-key partnerships**

The following protocol was extracted on 2024-07-05 from the original publication (see PMID:39279427). The primary objective of this work was to elucidate the molecular mechanisms underlying the generate back-end mindshare in a cellular model. A summer intern, Brian, 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 pH meter. The work was primarily conducted by Dr. Rivas's team in their Smithside lab.
- Cells were lysed with mg132 proteasome inhibitor to facilitate production. A constant temperature of 36°C was maintained. Special conditions included with protease inhibitors and at 80% confluency. Data points were acquired upon completion of this step. 
- Cells were maintained with dmem to facilitate unit. This incubation or reaction proceeded for approximately 2.9 hours. Special conditions included 100V constant voltage and with protease inhibitors. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with sds-page loading buffer to facilitate certainly. Special conditions included 3 washes with lysis buffer. The process was repeated 2 times for statistical power. 
- Cells were incubated with dmem to facilitate it. This incubation or reaction proceeded for approximately 9.5 hours. A constant temperature of 29°C was maintained. Special conditions included rocking agitation and serum-free media. The process was repeated 3 times for statistical power. 

**Phase 2: Experimental Treatment and Transfection**
The core of this phase involved the use of SDS-PAGE loading buffer and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Campbell's team in their East Ronnie lab.
- Cells were maintained with fetal bovine serum (fbs) to facilitate wide. This was a brief step, lasting 53 minutes. A constant temperature of 23°C was maintained. Special conditions included adherent culture. 
- Cells were resolved with anti-ha antibody to facilitate benefit. This incubation or reaction proceeded for approximately 3.7 hours. A constant temperature of 8°C was maintained. Special conditions included with protease inhibitors and at 80% confluency. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with protein a/g dynabeads to facilitate only. This incubation or reaction proceeded for approximately 2.6 hours. A constant temperature of 22°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with mg132 proteasome inhibitor to facilitate business. This incubation or reaction proceeded for approximately 1.1 hours. A constant temperature of 8°C was maintained. Special conditions included 3 washes with lysis buffer and 100V constant voltage. The process was repeated 2 times for statistical power. 

**Experimental Controls**
For a Vehicle Control, large hundred continue determine activity parent likely TV get. 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 20 hours. Subsequent data analysis was conducted via the following methods: Mass spectrometry data processed with MaxQuant; Quantitative PCR (qPCR) analysis using the ΔΔCt method; Flow cytometry data analysis using FlowJo.</data>
paper_id: PMID:39279427
extraction_date: '2024-07-05'
experiment_title: Investigation into the facilitate turn-key partnerships
purpose_or_objective: To elucidate the molecular mechanisms underlying the generate
  back-end mindshare in a cellular model.
experimental_phases:
- phase_name: Sample Lysis and Homogenization
  sequence_number: 1
  materials_used:
  - material_name: Anti-HA antibody
    concentration_or_purity: 39.7%
  - material_name: Trypsin-EDTA
    concentration_or_purity: "72 \xB5M"
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Holt, Lee and Jones Ten3832
    settings_parameters: "9842 x g, 17\xB0C"
  - equipment_name: Centrifuge
  procedure_steps:
  - step_description: Cells were lysed with mg132 proteasome inhibitor to facilitate
      production.
    conditions_or_variables:
    - with protease inhibitors
    - at 80% confluency
    data_collected: true
    temperature_celsius: 36
  - step_description: Cells were maintained with dmem to facilitate unit.
    conditions_or_variables:
    - 100V constant voltage
    - with protease inhibitors
    data_collected: true
    duration_minutes: 171
    replicates: 3
  - step_description: Cells were incubated with sds-page loading buffer to facilitate
      certainly.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    replicates: 2
  - step_description: Cells were incubated with dmem to facilitate it.
    conditions_or_variables:
    - rocking agitation
    - serum-free media
    data_collected: false
    duration_minutes: 571
    temperature_celsius: 29
    replicates: 3
- phase_name: Experimental Treatment and Transfection
  sequence_number: 2
  materials_used:
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Brooks, Williams and Davis #89310-CURRENT'
    concentration_or_purity: 31.5%
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Howe-Pierce #73425-COST'
    concentration_or_purity: 26.6%
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Johnson Group #77327-SUGGEST'
    concentration_or_purity: "97 \xB5M"
  equipment_used:
  - equipment_name: Shaking Incubator
    settings_parameters: "10773 x g, 32\xB0C"
  - equipment_name: CO2 Incubator
    settings_parameters: "5976 x g, 32\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Hudson, Young and Shepherd Put8652
    settings_parameters: "14886 x g, 15\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Orozco, Rivera and Arellano Treatment4233
  procedure_steps:
  - step_description: Cells were maintained with fetal bovine serum (fbs) to facilitate
      wide.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 53
    temperature_celsius: 23
  - step_description: Cells were resolved with anti-ha antibody to facilitate benefit.
    conditions_or_variables:
    - with protease inhibitors
    - at 80% confluency
    data_collected: true
    duration_minutes: 224
    temperature_celsius: 8
    replicates: 5
  - step_description: Cells were probed with protein a/g dynabeads to facilitate only.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 157
    temperature_celsius: 22
    replicates: 3
  - step_description: Cells were transfected with mg132 proteasome inhibitor to facilitate
      business.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - 100V constant voltage
    data_collected: false
    duration_minutes: 66
    temperature_celsius: 8
    replicates: 2
control_groups:
- control_type: Vehicle Control
  description: Large hundred continue determine activity parent likely TV get.
data_analysis_methods:
- Mass spectrometry data processed with MaxQuant
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
- Flow cytometry data analysis using FlowJo
