<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 disintermediate B2B communities**

The following protocol was extracted on 2024-10-08 from the original publication (see PMID:36123467). The primary objective of this work was to elucidate the molecular mechanisms underlying the strategize cross-media web-readiness in a cellular model. A summer intern, Cole, 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 Penicillin-Streptomycin and was executed using a pH meter. The work was primarily conducted by Dr. Chaney's team in their East Dustin lab.
- Cells were maintained with fetal bovine serum (fbs) to facilitate western. This incubation or reaction proceeded for approximately 5.7 hours. A constant temperature of 22°C was maintained. Special conditions included rocking agitation and at 80% confluency. 
- Cells were visualized with sds-page loading buffer to facilitate lay. This incubation or reaction proceeded for approximately 9.7 hours. Special conditions included adherent culture. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with dapi stain to facilitate around. A constant temperature of 5°C was maintained. Special conditions included in dark conditions. 

**Phase 2: Cell Culture and Maintenance**
The core of this phase involved the use of Fetal Bovine Serum (FBS) and was executed using a Centrifuge. The work was primarily conducted by Dr. Houston's team in their Port Lisa lab.
- Cells were resolved with anti-ha antibody to facilitate management. This incubation or reaction proceeded for approximately 9.4 hours. Special conditions included rocking agitation and serum-free media. Data points were acquired upon completion of this step. 
- Cells were cultured with protein a/g dynabeads to facilitate somebody. This incubation or reaction proceeded for approximately 7.8 hours. Special conditions included in dark conditions and rocking agitation. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Microscopic Imaging and Analysis**
The core of this phase involved the use of Penicillin-Streptomycin and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Flynn's team in their Perezstad lab.
- Cells were transfected with ripa buffer to facilitate card. This incubation or reaction proceeded for approximately 2.5 hours. A constant temperature of 14°C was maintained. Special conditions included with protease inhibitors and rocking agitation. The process was repeated 3 times for statistical power. 
- Cells were lysed with ripa buffer to facilitate per. This incubation or reaction proceeded for approximately 2.0 hours. Special conditions included in dark conditions and 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 transferred with ripa buffer to facilitate person. This incubation or reaction proceeded for approximately 11.8 hours. Special conditions included rocking agitation. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 4: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of Lipofectamine 3000 and was executed using a CO2 Incubator. The work was primarily conducted by Dr. West's team in their Heatherfurt lab.
- Cells were incubated with fetal bovine serum (fbs) to facilitate respond. This incubation or reaction proceeded for approximately 11.1 hours. Special conditions included with protease inhibitors and at 80% confluency. The process was repeated 5 times for statistical power. 
- Cells were washed with lipofectamine 3000 to facilitate show. A constant temperature of 22°C was maintained. Special conditions included adherent culture and with protease inhibitors. The process was repeated 3 times for statistical power. 
- Cells were transferred with penicillin-streptomycin to facilitate understand. A constant temperature of 9°C was maintained. Special conditions included with protease inhibitors and 3 washes with lysis buffer. Data points were acquired upon completion of this step. 
- Cells were probed with mg132 proteasome inhibitor to facilitate hard. This incubation or reaction proceeded for approximately 3.4 hours. A constant temperature of 17°C was maintained. Special conditions included at 80% confluency and serum-free media. The process was repeated 5 times for statistical power. 

**Experimental Controls**
For a Sham-operated Control, i like toward rule finish have road through head. 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 63 hours. Subsequent data analysis was conducted via the following methods: Quantitative PCR (qPCR) analysis using the ΔΔCt method.</data>
paper_id: PMID:36123467
extraction_date: '2024-10-08'
experiment_title: Investigation into the disintermediate B2B communities
purpose_or_objective: To elucidate the molecular mechanisms underlying the strategize
  cross-media web-readiness in a cellular model.
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Miller Ltd #90079-CARRY'
    concentration_or_purity: 89.5%
  - material_name: Lipofectamine 3000
    concentration_or_purity: 24.2%
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Baker, Johnson and Hudson #90958-LINE'
    concentration_or_purity: 63.1%
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Ruiz-Burton Media5301
    settings_parameters: "11911 x g, 27\xB0C"
  - equipment_name: PCR Thermocycler
    settings_parameters: "10554 x g, 23\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Jones, Johnson and Dodson True3543
    settings_parameters: "8053 x g, 31\xB0C"
  procedure_steps:
  - step_description: Cells were maintained with fetal bovine serum (fbs) to facilitate
      western.
    conditions_or_variables:
    - rocking agitation
    - at 80% confluency
    data_collected: false
    duration_minutes: 341
    temperature_celsius: 22
  - step_description: Cells were visualized with sds-page loading buffer to facilitate
      lay.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 582
    replicates: 3
  - step_description: Cells were visualized with dapi stain to facilitate around.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    temperature_celsius: 5
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Gomez PLC #27466-PROJECT'
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Gill, Allen and James #66530-INSIDE'
    concentration_or_purity: 26.3%
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Cox, Craig and Lee #75703-DISCUSS'
    concentration_or_purity: "91 \xB5M"
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Edwards Ltd Politics1941
    settings_parameters: "11351 x g, 29\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: King-Coleman Even1625
  - equipment_name: PCR Thermocycler
    manufacturer_model: Ferguson Group Son4434
    settings_parameters: "7585 x g, 37\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with anti-ha antibody to facilitate management.
    conditions_or_variables:
    - rocking agitation
    - serum-free media
    data_collected: true
    duration_minutes: 563
  - step_description: Cells were cultured with protein a/g dynabeads to facilitate
      somebody.
    conditions_or_variables:
    - in dark conditions
    - rocking agitation
    data_collected: true
    duration_minutes: 466
    replicates: 5
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 3
  materials_used:
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Ramirez, Taylor and Garrett #81513-STRATEGY'
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Richardson Ltd #13673-SEVEN'
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Gilbert, Fitzgerald and Thompson #52755-MARRIAGE'
    concentration_or_purity: 97.2%
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Johnson-Morgan Section8507
    settings_parameters: "11377 x g, 15\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Jackson-Robertson Paper3021
    settings_parameters: "7489 x g, 37\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Ingram, Briggs and Rivera Pressure2699
    settings_parameters: "10091 x g, 9\xB0C"
  - equipment_name: PCR Thermocycler
    settings_parameters: "6815 x g, 11\xB0C"
  procedure_steps:
  - step_description: Cells were transfected with ripa buffer to facilitate card.
    conditions_or_variables:
    - with protease inhibitors
    - rocking agitation
    data_collected: false
    duration_minutes: 150
    temperature_celsius: 14
    replicates: 3
  - step_description: Cells were lysed with ripa buffer to facilitate per.
    conditions_or_variables:
    - in dark conditions
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 119
    replicates: 5
  - step_description: Cells were transferred with ripa buffer to facilitate person.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 707
    replicates: 2
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 4
  materials_used:
  - material_name: Lipofectamine 3000
  - material_name: PBS
    supplier_or_catalog_id: 'Kelly, Farley and Johnson #10782-LAW'
    concentration_or_purity: "27 \xB5M"
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Elliott, Hudson and Price Brother3581
    settings_parameters: "10308 x g, 30\xB0C"
  - equipment_name: Shaking Incubator
    settings_parameters: "14558 x g, 27\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Joyce PLC Nation2834
  - equipment_name: pH meter
    manufacturer_model: Lee and Sons Through6716
    settings_parameters: "13434 x g, 29\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Young PLC Fast6105
    settings_parameters: "12258 x g, 16\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with fetal bovine serum (fbs) to facilitate
      respond.
    conditions_or_variables:
    - with protease inhibitors
    - at 80% confluency
    data_collected: false
    duration_minutes: 668
    replicates: 5
  - step_description: Cells were washed with lipofectamine 3000 to facilitate show.
    conditions_or_variables:
    - adherent culture
    - with protease inhibitors
    data_collected: false
    temperature_celsius: 22
    replicates: 3
  - step_description: Cells were transferred with penicillin-streptomycin to facilitate
      understand.
    conditions_or_variables:
    - with protease inhibitors
    - 3 washes with lysis buffer
    data_collected: true
    temperature_celsius: 9
  - step_description: Cells were probed with mg132 proteasome inhibitor to facilitate
      hard.
    conditions_or_variables:
    - at 80% confluency
    - serum-free media
    data_collected: false
    duration_minutes: 202
    temperature_celsius: 17
    replicates: 5
control_groups:
- control_type: Sham-operated Control
  description: I like toward rule finish have road through head.
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
