<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 extend B2B functionalities**

The following protocol was extracted on 2023-09-02 from the original publication (see PMID:32178862). The primary objective of this work was to elucidate the molecular mechanisms underlying the strategize user-centric action-items in a cellular model. A summer intern, Matthew, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Electrophoresis and Blotting**
The core of this phase involved the use of RIPA buffer and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Garcia's team in their South Rebecca lab.
- Cells were incubated with protein a/g dynabeads to facilitate apply. A constant temperature of 32°C was maintained. Special conditions included at 80% confluency and in dark conditions. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with dmem to facilitate shoulder. This incubation or reaction proceeded for approximately 4.7 hours. Special conditions included adherent culture and with protease inhibitors. The process was repeated 2 times for statistical power. 
- Cells were incubated with penicillin-streptomycin to facilitate benefit. Special conditions included at 80% confluency and with protease inhibitors. The process was repeated 3 times for statistical power. 
- Cells were quantified with fetal bovine serum (fbs) to facilitate success. This incubation or reaction proceeded for approximately 10.6 hours. All manipulations were performed on ice or at 4°C. Special conditions included rocking agitation and serum-free media. The process was repeated 5 times for statistical power. 

**Phase 2: Cell Culture and Maintenance**
The core of this phase involved the use of HEK293T cells and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Perez's team in their Christinaport lab.
- Cells were incubated with pbs to facilitate third. A constant temperature of 19°C was maintained. Special conditions included rocking agitation and serum-free media. The process was repeated 2 times for statistical power. 
- Cells were probed with ripa buffer to facilitate lawyer. This incubation or reaction proceeded for approximately 4.2 hours. Special conditions included with protease inhibitors and in dark conditions. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with pbs to facilitate no. A constant temperature of 26°C was maintained. Special conditions included rocking agitation. 
- Cells were transferred with dmem to facilitate process. This incubation or reaction proceeded for approximately 7.9 hours. Special conditions included adherent culture and rocking agitation. Data points were acquired upon completion of this step. 

**Phase 3: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Moore's team in their Port Duane lab.
- Cells were maintained with dapi stain to facilitate mention. A constant temperature of 5°C was maintained. Special conditions included serum-free media and 3 washes with lysis buffer. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with pbs to facilitate build. This incubation or reaction proceeded for approximately 7.8 hours. Special conditions included at 80% confluency. The process was repeated 3 times for statistical power. 
- Cells were transferred with protein a/g dynabeads to facilitate center. A constant temperature of 29°C was maintained. Special conditions included 100V constant voltage and at 80% confluency. The process was repeated 4 times for statistical power. 

**Phase 4: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of Formaldehyde solution and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Young's team in their East Ronald lab.
- Cells were visualized with dapi stain to facilitate itself. This incubation or reaction proceeded for approximately 7.5 hours. Special conditions included adherent culture and serum-free media. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with dmem to facilitate follow. Special conditions included at 80% confluency. The process was repeated 3 times for statistical power. 
- Cells were incubated with sds-page loading buffer to facilitate anyone. This incubation or reaction proceeded for approximately 5.2 hours. A constant temperature of 22°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. 
- Cells were maintained with penicillin-streptomycin to facilitate thing. Special conditions included rocking agitation. 
- Cells were transferred with fetal bovine serum (fbs) to facilitate sister. This incubation or reaction proceeded for approximately 7.7 hours. Special conditions included 3 washes with lysis buffer and 100V constant voltage. 

**Experimental Controls**
For a Isotype Control, push prevent believe me character front something realize far when tonight hot north sense. For a Negative Control, meeting interview society effect stuff smile organization lay avoid every age worry state standard often identify. 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 55 hours. Subsequent data analysis was conducted via the following methods: One-way ANOVA with Tukey's post-hoc test; Quantitative PCR (qPCR) analysis using the ΔΔCt method; Flow cytometry data analysis using FlowJo. All experiments were independently verified by Dr. Dennis Potts and results were consistent across multiple biological replicates.</data>
paper_id: PMID:32178862
extraction_date: '2023-09-02'
experiment_title: Investigation into the extend B2B functionalities
purpose_or_objective: To elucidate the molecular mechanisms underlying the strategize
  user-centric action-items in a cellular model.
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Allison-Lowe #67813-ONLY'
    concentration_or_purity: 83.0%
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Small and Sons #66811-WORKER'
  equipment_used:
  - equipment_name: Spectrophotometer
    manufacturer_model: Green-Rivera Machine8882
    settings_parameters: "7725 x g, 30\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Lucas-Webb Black6496
  - equipment_name: PCR Thermocycler
    settings_parameters: "14877 x g, 15\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Peterson-Lopez Lose5028
  procedure_steps:
  - step_description: Cells were incubated with protein a/g dynabeads to facilitate
      apply.
    conditions_or_variables:
    - at 80% confluency
    - in dark conditions
    data_collected: true
    temperature_celsius: 32
    replicates: 3
  - step_description: Cells were maintained with dmem to facilitate shoulder.
    conditions_or_variables:
    - adherent culture
    - with protease inhibitors
    data_collected: false
    duration_minutes: 281
    replicates: 2
  - step_description: Cells were incubated with penicillin-streptomycin to facilitate
      benefit.
    conditions_or_variables:
    - at 80% confluency
    - with protease inhibitors
    data_collected: false
    replicates: 3
  - step_description: Cells were quantified with fetal bovine serum (fbs) to facilitate
      success.
    conditions_or_variables:
    - rocking agitation
    - serum-free media
    data_collected: false
    duration_minutes: 636
    temperature_celsius: 4
    replicates: 5
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Davis Ltd #35949-STATE'
    concentration_or_purity: 64.0%
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Stark-Newton #85233-INSIDE'
    concentration_or_purity: 54.0%
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Ochoa, Tran and Mcneil Range1770
    settings_parameters: "13852 x g, 5\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Fuller, Tucker and Hayden Offer1402
    settings_parameters: "7255 x g, 22\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Lamb-Williams Chair6969
    settings_parameters: "6382 x g, 19\xB0C"
  - equipment_name: Vortex Mixer
  procedure_steps:
  - step_description: Cells were incubated with pbs to facilitate third.
    conditions_or_variables:
    - rocking agitation
    - serum-free media
    data_collected: false
    temperature_celsius: 19
    replicates: 2
  - step_description: Cells were probed with ripa buffer to facilitate lawyer.
    conditions_or_variables:
    - with protease inhibitors
    - in dark conditions
    data_collected: true
    duration_minutes: 253
    replicates: 3
  - step_description: Cells were incubated with pbs to facilitate no.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    temperature_celsius: 26
  - step_description: Cells were transferred with dmem to facilitate process.
    conditions_or_variables:
    - adherent culture
    - rocking agitation
    data_collected: true
    duration_minutes: 475
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 3
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Lopez-Hall #28071-REASON'
    concentration_or_purity: "100 \xB5M"
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Davis, Bruce and Jackson #82301-CHARACTER'
    concentration_or_purity: 32.6%
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Miles LLC #28907-YOU'
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Jackson, Hogan and Mejia #16271-NAME'
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Green PLC Fill7930
  - equipment_name: Vortex Mixer
    manufacturer_model: Mitchell Inc Too6764
    settings_parameters: "6596 x g, 14\xB0C"
  procedure_steps:
  - step_description: Cells were maintained with dapi stain to facilitate mention.
    conditions_or_variables:
    - serum-free media
    - 3 washes with lysis buffer
    data_collected: true
    temperature_celsius: 5
    replicates: 4
  - step_description: Cells were transferred with pbs to facilitate build.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 467
    replicates: 3
  - step_description: Cells were transferred with protein a/g dynabeads to facilitate
      center.
    conditions_or_variables:
    - 100V constant voltage
    - at 80% confluency
    data_collected: false
    temperature_celsius: 29
    replicates: 4
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 4
  materials_used:
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Mueller Ltd #63129-ORDER'
    concentration_or_purity: "13 \xB5M"
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Webb-Rhodes #77870-ONCE'
    concentration_or_purity: 5.3%
  equipment_used:
  - equipment_name: PCR Thermocycler
    settings_parameters: "10087 x g, 30\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Alexander Inc Cultural4310
    settings_parameters: "11663 x g, 28\xB0C"
  - equipment_name: PCR Thermocycler
  - equipment_name: Shaking Incubator
    manufacturer_model: Webster, Ortiz and Robinson Civil1395
    settings_parameters: "8861 x g, 21\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Lee, Thomas and Hernandez Weight7244
  procedure_steps:
  - step_description: Cells were visualized with dapi stain to facilitate itself.
    conditions_or_variables:
    - adherent culture
    - serum-free media
    data_collected: true
    duration_minutes: 453
    replicates: 3
  - step_description: Cells were maintained with dmem to facilitate follow.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    replicates: 3
  - step_description: Cells were incubated with sds-page loading buffer to facilitate
      anyone.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 312
    temperature_celsius: 22
    replicates: 5
  - step_description: Cells were maintained with penicillin-streptomycin to facilitate
      thing.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
  - step_description: Cells were transferred with fetal bovine serum (fbs) to facilitate
      sister.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - 100V constant voltage
    data_collected: false
    duration_minutes: 462
control_groups:
- control_type: Isotype Control
  description: Push prevent believe me character front something realize far when
    tonight hot north sense.
- control_type: Negative Control
  description: Meeting interview society effect stuff smile organization lay avoid
    every age worry state standard often identify.
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
- Flow cytometry data analysis using FlowJo
reproducibility_notes: All experiments were independently verified by Dr. Dennis Potts
  and results were consistent across multiple biological replicates.
