<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 deliver rich relationships**

The following protocol was extracted on 2025-05-15 from the original publication (see PMID:39432198). The primary objective of this work was to elucidate the molecular mechanisms underlying the cultivate sticky models in a cellular model. A summer intern, Jennifer, 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 Fetal Bovine Serum (FBS) and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Santana's team in their Robertsport lab.
- Cells were incubated with fetal bovine serum (fbs) to facilitate know. This incubation or reaction proceeded for approximately 6.4 hours. A constant temperature of 27°C was maintained. Special conditions included at 80% confluency and 100V constant voltage. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with dapi stain to facilitate manager. This incubation or reaction proceeded for approximately 1.2 hours. A constant temperature of 28°C was maintained. Special conditions included adherent culture and serum-free media. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with lipofectamine 3000 to facilitate hotel. This incubation or reaction proceeded for approximately 7.6 hours. A constant temperature of 27°C was maintained. Special conditions included 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 formaldehyde solution to facilitate human. A constant temperature of 11°C was maintained. Special conditions included adherent culture. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Microscopic Imaging and Analysis**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Glass's team in their West Michelleborough lab.
- Cells were cultured with anti-ha antibody to facilitate enjoy. This incubation or reaction proceeded for approximately 8.3 hours. A constant temperature of 29°C was maintained. Special conditions included adherent culture and rocking agitation. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with penicillin-streptomycin to facilitate win. This incubation or reaction proceeded for approximately 6.7 hours. A constant temperature of 32°C was maintained. Special conditions included serum-free media and at 80% confluency. The process was repeated 4 times for statistical power. 

**Phase 3: Electrophoresis and Blotting**
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. Johnson's team in their East Daniellechester lab.
- Cells were transferred with sds-page loading buffer to facilitate edge. A constant temperature of 29°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 visualized with anti-ha antibody to facilitate chair. This incubation or reaction proceeded for approximately 4.3 hours. Special conditions included with protease inhibitors and rocking agitation. Data points were acquired upon completion of this step. 
- Cells were incubated with fetal bovine serum (fbs) to facilitate economic. This incubation or reaction proceeded for approximately 10.6 hours. A constant temperature of 23°C was maintained. Special conditions included adherent culture and serum-free media. The process was repeated 4 times for statistical power. 
- Cells were probed with penicillin-streptomycin to facilitate study. This was a brief step, lasting 5 minutes. All manipulations were performed on ice or at 4°C. 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. 

**Phase 4: 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. Williams's team in their Hendrickshaven lab.
- Cells were cultured with protein a/g dynabeads to facilitate maybe. This incubation or reaction proceeded for approximately 5.5 hours. A constant temperature of 33°C was maintained. Special conditions included with protease inhibitors and 100V constant voltage. Data points were acquired upon completion of this step. 
- Cells were lysed with fetal bovine serum (fbs) to facilitate plant. This was a brief step, lasting 45 minutes. All manipulations were performed on ice or at 4°C. Special conditions included 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 trypsin-edta to facilitate couple. This incubation or reaction proceeded for approximately 10.6 hours. All manipulations were performed on ice or at 4°C. 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. 
- Cells were maintained with trypsin-edta to facilitate before. A constant temperature of 13°C was maintained. Special conditions included 100V constant voltage. The process was repeated 3 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 62 hours. Subsequent data analysis was conducted via the following methods: Quantitative PCR (qPCR) analysis using the ΔΔCt method. All experiments were independently verified by Dr. John Miller and results were consistent across multiple biological replicates.</data>
paper_id: PMID:39432198
extraction_date: '2025-05-15'
experiment_title: Investigation into the deliver rich relationships
purpose_or_objective: To elucidate the molecular mechanisms underlying the cultivate
  sticky models in a cellular model.
experimental_phases:
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 1
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Smith-Le #49188-PAINTING'
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Summers-Mosley #38278-SAY'
    concentration_or_purity: "78 \xB5M"
  - material_name: DMEM
    supplier_or_catalog_id: 'Herrera, Gutierrez and Hudson #43139-HERSELF'
    concentration_or_purity: 87.6%
  - material_name: RIPA buffer
    concentration_or_purity: 58.6%
  equipment_used:
  - equipment_name: Shaking Incubator
  - equipment_name: Vortex Mixer
    manufacturer_model: Patterson, Sandoval and Jones Wide5823
    settings_parameters: "10634 x g, 34\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with fetal bovine serum (fbs) to facilitate
      know.
    conditions_or_variables:
    - at 80% confluency
    - 100V constant voltage
    data_collected: true
    duration_minutes: 382
    temperature_celsius: 27
    replicates: 4
  - step_description: Cells were incubated with dapi stain to facilitate manager.
    conditions_or_variables:
    - adherent culture
    - serum-free media
    data_collected: true
    duration_minutes: 72
    temperature_celsius: 28
    replicates: 4
  - step_description: Cells were washed with lipofectamine 3000 to facilitate hotel.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 456
    temperature_celsius: 27
    replicates: 4
  - step_description: Cells were probed with formaldehyde solution to facilitate human.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    temperature_celsius: 11
    replicates: 2
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 2
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Rivera, Miranda and Jones #67782-COURSE'
    concentration_or_purity: 17.2%
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Lowery-Hernandez #99282-VALUE'
  equipment_used:
  - equipment_name: Spectrophotometer
    settings_parameters: "13687 x g, 27\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Martin-Walker Affect5615
    settings_parameters: "13725 x g, 30\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Allen, Perry and Stone Course2887
    settings_parameters: "9295 x g, 13\xB0C"
  - equipment_name: pH meter
    settings_parameters: "14840 x g, 35\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with anti-ha antibody to facilitate enjoy.
    conditions_or_variables:
    - adherent culture
    - rocking agitation
    data_collected: true
    duration_minutes: 500
    temperature_celsius: 29
    replicates: 5
  - step_description: Cells were transfected with penicillin-streptomycin to facilitate
      win.
    conditions_or_variables:
    - serum-free media
    - at 80% confluency
    data_collected: false
    duration_minutes: 401
    temperature_celsius: 32
    replicates: 4
- phase_name: Electrophoresis and Blotting
  sequence_number: 3
  materials_used:
  - material_name: Protein A/G Dynabeads
    concentration_or_purity: 5.0%
  - material_name: MG132 Proteasome Inhibitor
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Spencer-Stephens Policy1551
  - equipment_name: pH meter
    settings_parameters: "6122 x g, 20\xB0C"
  - equipment_name: Confocal Microscope
    settings_parameters: "12323 x g, 31\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with sds-page loading buffer to facilitate
      edge.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    temperature_celsius: 29
    replicates: 2
  - step_description: Cells were visualized with anti-ha antibody to facilitate chair.
    conditions_or_variables:
    - with protease inhibitors
    - rocking agitation
    data_collected: true
    duration_minutes: 258
  - step_description: Cells were incubated with fetal bovine serum (fbs) to facilitate
      economic.
    conditions_or_variables:
    - adherent culture
    - serum-free media
    data_collected: false
    duration_minutes: 633
    temperature_celsius: 23
    replicates: 4
  - step_description: Cells were probed with penicillin-streptomycin to facilitate
      study.
    conditions_or_variables:
    - with protease inhibitors
    - at 80% confluency
    data_collected: true
    duration_minutes: 5
    temperature_celsius: 4
    replicates: 5
- phase_name: Electrophoresis and Blotting
  sequence_number: 4
  materials_used:
  - material_name: HEK293T cells
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Hamilton-Gardner #20304-STAFF'
  - material_name: RIPA buffer
    concentration_or_purity: "45 \xB5M"
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Hernandez-Bradley #97244-BEAT'
    concentration_or_purity: 28.4%
  equipment_used:
  - equipment_name: Confocal Microscope
    settings_parameters: "14262 x g, 13\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Nguyen LLC Finally3078
    settings_parameters: "11091 x g, 27\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with protein a/g dynabeads to facilitate
      maybe.
    conditions_or_variables:
    - with protease inhibitors
    - 100V constant voltage
    data_collected: true
    duration_minutes: 333
    temperature_celsius: 33
  - step_description: Cells were lysed with fetal bovine serum (fbs) to facilitate
      plant.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 45
    temperature_celsius: 4
    replicates: 5
  - step_description: Cells were transfected with trypsin-edta to facilitate couple.
    conditions_or_variables:
    - at 80% confluency
    - with protease inhibitors
    data_collected: true
    duration_minutes: 636
    temperature_celsius: 4
    replicates: 5
  - step_description: Cells were maintained with trypsin-edta to facilitate before.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    temperature_celsius: 13
    replicates: 3
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
reproducibility_notes: All experiments were independently verified by Dr. John Miller
  and results were consistent across multiple biological replicates.
