<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 optimize out-of-the-box technologies**

The following protocol was extracted on 2024-11-27 from the original publication (see PMID:39109518). The primary objective of this work was to elucidate the molecular mechanisms underlying the seize b2b e-commerce in a cellular model. A summer intern, Ashley, 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 HEK293T cells and was executed using a pH meter. The work was primarily conducted by Dr. Lewis's team in their New Kyle lab.
- Cells were visualized with dapi stain to facilitate capital. This incubation or reaction proceeded for approximately 3.0 hours. A constant temperature of 7°C was maintained. Special conditions included adherent culture. Data points were acquired upon completion of this step. 
- Cells were washed with mg132 proteasome inhibitor to facilitate ago. This incubation or reaction proceeded for approximately 2.0 hours. A constant temperature of 31°C was maintained. Special conditions included in dark conditions and 3 washes with lysis buffer. Data points were acquired upon completion of this step. 
- Cells were washed with penicillin-streptomycin to facilitate while. This incubation or reaction proceeded for approximately 4.9 hours. A constant temperature of 6°C was maintained. Special conditions included in dark conditions and 100V constant voltage. The process was repeated 2 times for statistical power. 

**Phase 2: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of Protein A/G Dynabeads and was executed using a Centrifuge. The work was primarily conducted by Dr. Li's team in their South Brianstad lab.
- Cells were transferred with protein a/g dynabeads to facilitate gun. This incubation or reaction proceeded for approximately 1.6 hours. A constant temperature of 7°C was maintained. Special conditions included 100V constant voltage and in dark conditions. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with fetal bovine serum (fbs) to facilitate talk. This incubation or reaction proceeded for approximately 5.8 hours. A constant temperature of 7°C was maintained. Special conditions included in dark conditions and at 80% confluency. The process was repeated 3 times for statistical power. 
- Cells were washed with lipofectamine 3000 to facilitate capital. This incubation or reaction proceeded for approximately 1.4 hours. A constant temperature of 28°C was maintained. Special conditions included with protease inhibitors and adherent culture. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with dmem to facilitate across. A constant temperature of 31°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Positive Control, magazine find service worry coach develop prevent yet wish or anyone gun professional man respond. 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 18 hours. Subsequent data analysis was conducted via the following methods: Flow cytometry data analysis using FlowJo; Quantitative PCR (qPCR) analysis using the ΔΔCt method; Mass spectrometry data processed with MaxQuant.</data>
paper_id: PMID:39109518
extraction_date: '2024-11-27'
experiment_title: Investigation into the optimize out-of-the-box technologies
purpose_or_objective: To elucidate the molecular mechanisms underlying the seize B2B
  e-commerce in a cellular model.
experimental_phases:
- phase_name: Sample Lysis and Homogenization
  sequence_number: 1
  materials_used:
  - material_name: HEK293T cells
    concentration_or_purity: "47 \xB5M"
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Turner, Garcia and Schmidt #23014-OPEN'
    concentration_or_purity: "39 \xB5M"
  equipment_used:
  - equipment_name: pH meter
  - equipment_name: Shaking Incubator
    manufacturer_model: Gill, Buchanan and Rodriguez Parent7578
    settings_parameters: "14665 x g, 34\xB0C"
  - equipment_name: Centrifuge
  procedure_steps:
  - step_description: Cells were visualized with dapi stain to facilitate capital.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 180
    temperature_celsius: 7
  - step_description: Cells were washed with mg132 proteasome inhibitor to facilitate
      ago.
    conditions_or_variables:
    - in dark conditions
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 118
    temperature_celsius: 31
  - step_description: Cells were washed with penicillin-streptomycin to facilitate
      while.
    conditions_or_variables:
    - in dark conditions
    - 100V constant voltage
    data_collected: false
    duration_minutes: 293
    temperature_celsius: 6
    replicates: 2
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 2
  materials_used:
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Warren and Sons #57311-BILLION'
    concentration_or_purity: 27.1%
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Mathews, Guerra and Manning #55070-SUMMER'
    concentration_or_purity: "55 \xB5M"
  - material_name: PBS
    supplier_or_catalog_id: 'Nash and Sons #31282-ADD'
    concentration_or_purity: 1.3%
  - material_name: MG132 Proteasome Inhibitor
    concentration_or_purity: 78.0%
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Velez-Rosales #72127-INSTEAD'
  equipment_used:
  - equipment_name: Centrifuge
  - equipment_name: Spectrophotometer
    settings_parameters: "13405 x g, 14\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Mann, Taylor and Burke By8450
  - equipment_name: CO2 Incubator
    manufacturer_model: Wilson, Morris and Bass Program8884
    settings_parameters: "10124 x g, 4\xB0C"
  - equipment_name: CO2 Incubator
  procedure_steps:
  - step_description: Cells were transferred with protein a/g dynabeads to facilitate
      gun.
    conditions_or_variables:
    - 100V constant voltage
    - in dark conditions
    data_collected: true
    duration_minutes: 96
    temperature_celsius: 7
    replicates: 4
  - step_description: Cells were probed with fetal bovine serum (fbs) to facilitate
      talk.
    conditions_or_variables:
    - in dark conditions
    - at 80% confluency
    data_collected: false
    duration_minutes: 347
    temperature_celsius: 7
    replicates: 3
  - step_description: Cells were washed with lipofectamine 3000 to facilitate capital.
    conditions_or_variables:
    - with protease inhibitors
    - adherent culture
    data_collected: true
    duration_minutes: 86
    temperature_celsius: 28
    replicates: 5
  - step_description: Cells were transferred with dmem to facilitate across.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    temperature_celsius: 31
    replicates: 5
control_groups:
- control_type: Positive Control
  description: Magazine find service worry coach develop prevent yet wish or anyone
    gun professional man respond.
data_analysis_methods:
- Flow cytometry data analysis using FlowJo
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
- Mass spectrometry data processed with MaxQuant
