Western Blotting

BlockPRO™ Protein-Free Blocking Buffer
Protein-Free Blocking Buffer for Your Every Need.

BlockPRO™ Protein Free Blocking Buffer is a non-protein formulation which enhances sensitivity and minimizes background noise, presenting better results than traditional protein-based blocking buffer in immunoassays. The synthetic formulation of BlockPRO™ Protein-Free Blocking Buffer makes it suitable for PVDF and nitrocellulose platform, avidin/biotin system, detection of phosphoprotein, and other immunochemical applications.



  • High performance: Provide better specific signal and less background noise than traditional blocking buffer
  • Multiple application: Suitable for Western blot, dot blot, ELISA, and other immunoassays
  • High quality control: Ensure lot-to-lot consistency for your most reproducible results over time



Order Information:


Cat. No. Product Name Description
BF01-1L BlockPRO™ Protein-Free Blocking Buffer

500mL Solution X 2

BF10-100 BlockPRO™ Protein-Free Blocking Buffer (10X)

100mL 10X Solution X 1

BF10-200  BlockPRO™ Protein-Free Blocking Buffer (10X)

100mL 10X Solution X 2



Product Detail:



Figure 1. BlockPRO™ Protein-Free Blocking Buffer is better than protein-based blocking buffers (skim milk, BSA and casein) for detection of target protein in Western blotting.

THP-1 cell lysates were prepared and separated by electrophoresis. The proteins were transferred to PVDF and blocked for 1 hour at room temperature with the indicated blocking buffer, probed with mouse anti-pAMPK followed by anti-mouse HRP and detected by chemiluminescence. All results were exposed to X-ray film for 30 seconds.





Figure 2. BlockPRO™ Protein-Free is suitable for various protein antigen detection, such as high molecular weight protein, tACC; phosphoprotein, pAMPK; abundant protein, GAPDH; low molecular weight protein, histone H3.

THP-1 cell lysates were prepared and separated by electrophoresis. The proteins were transferred to PVDF and blocked for 1 hour at room temperature with BlockPRO™ Protein-Free Blocking Buffer. Antibodies designed to probe the indicated proteins were used. All the signals were detected by chemiluminescence and were exposed to X-ray film.





Figure  3. BlockPRO™ Protein-Free Blocking Buffer can be used in both PVDF and nitrocellulose platform. 

Hela cell lysates were prepared and separated by SDS-PAGE. The proteins were transferred to PVDF or nitrocellulose membranes. The membranes were blocked for overnight at 4 °C with BlockPRO™ Protein-Free Blocking Buffer or 5% skim milk, probed with mouse anti-histone H3 followed by anti-mouse HRP and detected by chemiluminescence. All results were exposed to X-ray film for 30 seconds.



Selection Guide:




1. Scheidler, Christopher M., Milan Vrabel, and Sabine Schneider. "Genetic Code Expansion, Protein Expression, and Protein Functionalization in Bacillus subtilis." ACS Synthetic Biology 9.3 (2020): 486-493. (ELISA)

2. CHEN, Kung‑Yen, et al. Monascin accelerates anoikis in circulating tumor cells and prevents breast cancer metastasis. Oncology Letters, 2020, 20.5: 1-1. (WB)

3. Chang, Shih‐Chieh, et al. "Significant association of serum autoantibodies to TYMS, HAPLN1 and IGFBP5 with early stage canine malignant mammary tumours." Veterinary and Comparative Oncology 19.1 (2021): 172-182. (ELISA)

4. Wang, Ruike, et al. “High-throughput immunosensor chip coupled with a fluorescent DNA dendrimer for ultrasensitive detection of cardiac troponin T.” RSC Advances 11.44 (2021): 27523-27529. (Immunosensor chip)

5. Lin, Chi-Chien, et al. "Crassolide Suppresses Dendritic Cell Maturation and Attenuates Experimental Antiphospholipid Syndrome." Molecules 26.9 (2021): 2492. (WB)

6. Tang, Kuo-Tung, et al. "Kurarinone Attenuates Collagen-Induced Arthritis in Mice by Inhibiting Th1/Th17 Cell Responses and Oxidative Stress." International journal of molecular sciences 22.8 (2021): 4002. (WB)

7. Zhang, Man-Li, Wei-Wei Liu, and Wei-Dong Li. “Imbalance of Molecular Module of TINCR-miR-761 Promotes the Metastatic Potential of Early Triple Negative Breast Cancer and Partially Offsets the Anti-Tumor Activity of Luteolin.” Cancer Management and Research 13 (2021): 1877. (WB)

8. Yuan, Stephen Hsien-Chi, et al. "Serum Level of Tumor-Overexpressed AGR2 is Significantly Associated with Unfavorable Prognosis of Canine Malignant Mammary Tumors." Animals 11.10 (2021): 2923. (ELISA, IF)

9. Chang, Kai-Wei, et al. “Atractylodin Suppresses TGF-β-Mediated Epithelial-Mesenchymal Transition in Alveolar Epithelial Cells and Attenuates Bleomycin-Induced Pulmonary Fibrosis in Mice.” International journal of molecular sciences 22.20 (2021): 11152. (WB)

10. Horikawa, Mei, Hisataka Sabe, and Yasuhito Onodera. “Dual roles of AMAP1 in the transcriptional regulation and intracellular trafficking of carbonic anhydrase IX.” Translational Oncology 15.1 (2022): 101258. (WB)

11. Lu, I-Ta, et al. “(−)-Agelasidine A Induces Endoplasmic Reticulum Stress-Dependent Apoptosis in Human Hepatocellular Carcinoma.” Marine Drugs 20.2 (2022): 109. (WB)

12. HM Yuki et al. "Molecular characterization of Vasa homolog in the pen shell Atrina pectinata: cDNA cloning and expression analysis during gonadal development." Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology (2022): 110798. (WB)

13. Chen, Ya-Hui, et al. "Synergistic combination of luteolin and asiatic acid on cervical cancer in vitro and in vivo." Cancers 15.2 (2023): 548. (WB)

14. Puangpila, Chanida, et al. "Development of lectin‐based lateral flow assay for fucosylated alpha‐fetoprotein." Journal of Cellular Biochemistry 124.10 (2023): 1546-1556. (Lateral flow assay)

15. Lim, Jie Lay, et al. "Curcumin-derived carbon quantum dots: Dual actions in mitigating tau hyperphosphorylation and amyloid beta aggregation." Colloids and Surfaces B: Biointerfaces 234 (2024): 113676. (WB)

16. Peala, Wisuttiya, et al. "Paper-based electrochemical immunosensor for highly sensitive detection of chicken anemia virus." Talanta (2024): 125820. (Immunoscensor)

17. Thanongsaksrikul, Jeeraphong, et al. "Characterization of antibody response to SARS-CoV-2 Orf8 from three waves of COVID-19 outbreak in Thailand." PloS one 19.5 (2024): e0297272.(ELISA)