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PROTEIN ANALYSIS

PROTEIN ANALYSIS SERVICE

As Nepenthe Research Technologies, the following services are provided within the scope of protein analysis. We can plan your project by selecting any of these services according to your needs.

  1. Protein quantification via Nanodrop
  2. BCA Assay
  3. SDS-PAGE
  4. Coomassie Staining
  5. Western Blotting

Subject

Details

Required Information
  • Functional role of the protein (if known)
  • Estimated concentration range (if known)
  • Sample type (liquid, lyophilized, cell pellet, tissue, etc.)
  • Sample buffer composition (Presence of detergents, reducing agents, etc. is critical for BCA and Nanodrop)
  • Molecular weight of the target protein (kDa) and NCBI/Uniprot ID (if known)
  • Other requirements: Special staining requests

For western blot:

  • Name of the antibody target
  • Type of antibody (e.g., polyclonal, monoclonal)
  • Source of the antibody (e.g., rabbit, goat, mouse)
  • Estimated purity and concentration of the antibody
  • Other requirements: labeling, conjugation, etc.

Protocol

1. Concentration and Purity Analysis:

  • Nanodrop: Rapid quantification via A280 absorbance and nucleic acid contamination check (A260/A280 ratio).
  • BCA Assay: Precise total protein quantification using a colorimetric method (562 nm) with high detergent tolerance, based on BSA standard curve.

2. Visualization and Separation (SDS-PAGE & Coomassie):

  • Selection of appropriate gel percentage (cast or pre-cast) for the protein's molecular weight.
  • Electrophoresis under reducing or non-reducing conditions.
  • Coomassie Blue Staining: Visualization of total protein profile and visual analysis of purity.

3. Specific Identification (Western Blot):

  • Transfer of proteins from gel to membrane (PVDF or Nitrocellulose).
  • Specific band detection via target-specific antibody incubation and chemiluminescence imaging.

Quality Control
  • Standard curve linearity check for BCA assays (R² > 0.98).
  • Verification of molecular weight markers (Ladder) for SDS-PAGE and Western Blot.
  • Normalization with loading control (e.g., Beta-actin, GAPDH) in Western Blot analyses (if requested).

Time
  • Basic Analyses (Nanodrop/BCA): 1 week
  • Gel and Blot Analyses: 2 weeks (May vary depending on the number of samples and antibody procurement process)

Provided Products
  • Detailed Analysis Report (including all experimental parameters)
  • High-resolution gel and blot images (TIF/JPEG format)

Price
  • Prices may vary depending on the number of samples, requested analysis combination (e.g., BCA only or Full Western Blot), and antibody availability. Please contact us for pricing information.

Others
  • We offer buffer exchange or concentration (ultrafiltration) services if required before analysis. We also provide semi-quantitative analysis support using ImageJ/ImageLab software on imaged gels. For more information, please contact us.

 

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