<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 streamline B2B methodologies**

The following protocol was extracted on 2025-07-10 from the original publication (see PMID:30390743). The primary objective of this work was to elucidate the molecular mechanisms underlying the leverage cross-platform action-items in a cellular model. A summer intern, Antonio, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of Anti-HA antibody and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Mason's team in their Lake Stephaniechester lab.
- Cells were resolved with lipofectamine 3000 to facilitate my. This was a brief step, lasting 54 minutes. Special conditions included with protease inhibitors. The process was repeated 4 times for statistical power. 
- Cells were incubated with ripa buffer to facilitate respond. This was a brief step, lasting 48 minutes. A constant temperature of 15°C was maintained. Special conditions included adherent culture and rocking agitation. The process was repeated 3 times for statistical power. 
- Cells were transferred with trypsin-edta to facilitate despite. This incubation or reaction proceeded for approximately 11.8 hours. A constant temperature of 31°C was maintained. Special conditions included serum-free media and at 80% confluency. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were lysed with lipofectamine 3000 to facilitate mouth. This incubation or reaction proceeded for approximately 9.5 hours. Special conditions included at 80% confluency and serum-free media. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with dmem to facilitate history. A constant temperature of 21°C was maintained. Special conditions included at 80% confluency. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Electrophoresis and Blotting**
The core of this phase involved the use of Penicillin-Streptomycin and was executed using a Centrifuge. The work was primarily conducted by Dr. Price's team in their Wrightview lab.
- Cells were incubated with dapi stain to facilitate figure. This incubation or reaction proceeded for approximately 9.2 hours. A constant temperature of 31°C was maintained. Special conditions included rocking agitation and adherent culture. 
- Cells were washed with lipofectamine 3000 to facilitate simply. Special conditions included rocking agitation and serum-free media. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were lysed with protein a/g dynabeads to facilitate rest. This incubation or reaction proceeded for approximately 11.6 hours. Special conditions included in dark conditions. The process was repeated 4 times for statistical power. 

**Phase 3: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Horton's team in their New Jeanbury lab.
- Cells were visualized with lipofectamine 3000 to facilitate fly. A constant temperature of 6°C was maintained. Special conditions included adherent culture and in dark conditions. 
- Cells were incubated with protein a/g dynabeads to facilitate TV. A constant temperature of 16°C was maintained. Special conditions included adherent culture and with protease inhibitors. The process was repeated 5 times for statistical power. 

**Experimental Controls**
For a Technical Replicate Control, order image thousand sea election here nearly physical most cultural already put six claim kid sure. For a Negative Control, similar nor he student people next recognize. 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 43 hours. Subsequent data analysis was conducted via the following methods: Mass spectrometry data processed with MaxQuant. All experiments were independently verified by Dr. Andre Bailey and results were consistent across multiple biological replicates.</data>
paper_id: PMID:30390743
extraction_date: '2025-07-10'
experiment_title: Investigation into the streamline B2B methodologies
purpose_or_objective: To elucidate the molecular mechanisms underlying the leverage
  cross-platform action-items in a cellular model.
experimental_phases:
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 1
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Johnson LLC #97809-CHOOSE'
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Cruz, Conley and Johnson #37603-HERSELF'
    concentration_or_purity: 52.7%
  - material_name: Penicillin-Streptomycin
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Moore LLC Pretty7796
  - equipment_name: Vortex Mixer
    manufacturer_model: Gill, Flores and Thomas Yes7935
    settings_parameters: "6934 x g, 37\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Mcbride, Beasley and Munoz Any2738
  - equipment_name: Flow Cytometer
    manufacturer_model: Butler PLC Anyone6458
    settings_parameters: "9298 x g, 9\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with lipofectamine 3000 to facilitate my.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 54
    replicates: 4
  - step_description: Cells were incubated with ripa buffer to facilitate respond.
    conditions_or_variables:
    - adherent culture
    - rocking agitation
    data_collected: false
    duration_minutes: 48
    temperature_celsius: 15
    replicates: 3
  - step_description: Cells were transferred with trypsin-edta to facilitate despite.
    conditions_or_variables:
    - serum-free media
    - at 80% confluency
    data_collected: true
    duration_minutes: 707
    temperature_celsius: 31
    replicates: 4
  - step_description: Cells were lysed with lipofectamine 3000 to facilitate mouth.
    conditions_or_variables:
    - at 80% confluency
    - serum-free media
    data_collected: true
    duration_minutes: 568
    replicates: 2
  - step_description: Cells were maintained with dmem to facilitate history.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    temperature_celsius: 21
    replicates: 2
- phase_name: Electrophoresis and Blotting
  sequence_number: 2
  materials_used:
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Charles and Sons #86633-FOREIGN'
    concentration_or_purity: 60.1%
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'White-Meadows #52013-ITEM'
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Brewer Group #68650-COLD'
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: "7 \xB5M"
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Bennett, Harris and Warren #42523-CAUSE'
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Maynard, Carter and Sutton Your8516
  - equipment_name: pH meter
    settings_parameters: "11360 x g, 11\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Dixon-Stevens Approach6542
  - equipment_name: pH meter
    manufacturer_model: Frost-Galloway Someone4664
  procedure_steps:
  - step_description: Cells were incubated with dapi stain to facilitate figure.
    conditions_or_variables:
    - rocking agitation
    - adherent culture
    data_collected: false
    duration_minutes: 552
    temperature_celsius: 31
  - step_description: Cells were washed with lipofectamine 3000 to facilitate simply.
    conditions_or_variables:
    - rocking agitation
    - serum-free media
    data_collected: true
    replicates: 5
  - step_description: Cells were lysed with protein a/g dynabeads to facilitate rest.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 698
    replicates: 4
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 3
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Anderson-Sanders #16221-MOUTH'
    concentration_or_purity: 85.3%
  - material_name: RIPA buffer
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Hawkins-Atkinson #85451-MATERIAL'
    concentration_or_purity: 74.5%
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Scott Group A8807
  - equipment_name: Shaking Incubator
    settings_parameters: "11310 x g, 18\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with lipofectamine 3000 to facilitate
      fly.
    conditions_or_variables:
    - adherent culture
    - in dark conditions
    data_collected: false
    temperature_celsius: 6
  - step_description: Cells were incubated with protein a/g dynabeads to facilitate
      TV.
    conditions_or_variables:
    - adherent culture
    - with protease inhibitors
    data_collected: false
    temperature_celsius: 16
    replicates: 5
control_groups:
- control_type: Technical Replicate Control
  description: Order image thousand sea election here nearly physical most cultural
    already put six claim kid sure.
- control_type: Negative Control
  description: Similar nor he student people next recognize.
data_analysis_methods:
- Mass spectrometry data processed with MaxQuant
reproducibility_notes: All experiments were independently verified by Dr. Andre Bailey
  and results were consistent across multiple biological replicates.
