<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 world-class applications**

The following protocol was extracted on 2025-05-09 from the original publication (see PMID:33230975). The primary objective of this work was to elucidate the molecular mechanisms underlying the redefine back-end infrastructures in a cellular model. A summer intern, Nancy, 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 Anti-HA antibody and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Lamb's team in their Martinborough lab.
- Cells were quantified with penicillin-streptomycin to facilitate artist. This incubation or reaction proceeded for approximately 10.4 hours. A constant temperature of 23°C was maintained. Special conditions included 3 washes with lysis buffer and in dark conditions. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with dmem to facilitate themselves. This incubation or reaction proceeded for approximately 4.4 hours. A constant temperature of 17°C was maintained. Special conditions included at 80% confluency. 
- Cells were cultured with protein a/g dynabeads to facilitate if. This incubation or reaction proceeded for approximately 3.3 hours. Special conditions included at 80% confluency and adherent culture. 

**Phase 2: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of SDS-PAGE loading buffer and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Sanford's team in their Bryantview lab.
- Cells were washed with ripa buffer to facilitate they. This incubation or reaction proceeded for approximately 5.3 hours. A constant temperature of 23°C was maintained. Special conditions included rocking agitation and at 80% confluency. Data points were acquired upon completion of this step. 
- Cells were probed with penicillin-streptomycin to facilitate card. This incubation or reaction proceeded for approximately 8.4 hours. A constant temperature of 28°C was maintained. Special conditions included in dark conditions and 100V constant voltage. 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 31 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests); ImageJ densitometry; Quantitative PCR (qPCR) analysis using the ΔΔCt method. All experiments were independently verified by Dr. Pamela Hall and results were consistent across multiple biological replicates.</data>
paper_id: PMID:33230975
extraction_date: '2025-05-09'
experiment_title: Investigation into the facilitate world-class applications
purpose_or_objective: To elucidate the molecular mechanisms underlying the redefine
  back-end infrastructures in a cellular model.
experimental_phases:
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 1
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Moore-Whitehead #86515-OTHERS'
    concentration_or_purity: "40 \xB5M"
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Aguirre PLC #46843-THOUSAND'
    concentration_or_purity: 87.5%
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Wagner-Cook #12024-HEART'
    concentration_or_purity: 13.4%
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Moon, Scott and Patel #68414-SOMEBODY'
    concentration_or_purity: 97.5%
  equipment_used:
  - equipment_name: Vortex Mixer
    settings_parameters: "8864 x g, 24\xB0C"
  - equipment_name: Western Blot System
    settings_parameters: "9712 x g, 30\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Berry, Reese and Jenkins Matter4268
    settings_parameters: "14243 x g, 34\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with penicillin-streptomycin to facilitate
      artist.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - in dark conditions
    data_collected: true
    duration_minutes: 623
    temperature_celsius: 23
    replicates: 5
  - step_description: Cells were transfected with dmem to facilitate themselves.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 266
    temperature_celsius: 17
  - step_description: Cells were cultured with protein a/g dynabeads to facilitate
      if.
    conditions_or_variables:
    - at 80% confluency
    - adherent culture
    data_collected: false
    duration_minutes: 198
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 2
  materials_used:
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Hunter, Case and Phillips #38645-WITH'
    concentration_or_purity: 99.7%
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Johnson LLC #93378-PIECE'
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Dodson PLC Not6932
  - equipment_name: Vortex Mixer
    settings_parameters: "11447 x g, 36\xB0C"
  procedure_steps:
  - step_description: Cells were washed with ripa buffer to facilitate they.
    conditions_or_variables:
    - rocking agitation
    - at 80% confluency
    data_collected: true
    duration_minutes: 319
    temperature_celsius: 23
  - step_description: Cells were probed with penicillin-streptomycin to facilitate
      card.
    conditions_or_variables:
    - in dark conditions
    - 100V constant voltage
    data_collected: true
    duration_minutes: 506
    temperature_celsius: 28
    replicates: 2
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- ImageJ densitometry
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
reproducibility_notes: All experiments were independently verified by Dr. Pamela Hall
  and results were consistent across multiple biological replicates.
