Blocking Buffer for Fluorescent Western Blotting

Catalog No. ABIN925618

Product Details

Application: Blocking Reagent (BR), Labeling (Lbl), Western Blotting (WB)

Purpose: Blocking Buffer for Fluorescent Western Blotting

Specificity: Blocking buffer was prepared using ultra pure reagents dissolved in pharmaceutical grade water (WFI) and consists of a proprietary protein formulation in TRIS buffered saline at pH 7.6 with thimerosal added as an antimicrobial agent.

Characteristics: Western Blot Blocking Solution is specifically designed for western blotting using fluorochrome conjugated antibodies. Pure nitrocellulose membrane is recommended for maximum performance. Other membranes, such as PVDF or nitrocellulose embedded in a support can be used, but may generate elevated backgrounds. Protein should be transferred from gel to membrane using standard protocols.
Blocking buffer can be used for membrane blocking and to dilute both primary and secondary antibodies.
Western Blot blocking buffer is suitable for use with fluorescent western blot imaging systems produced by Bio-Rad Laboratories, GE Healthcare, Alpha Innotech, FujiFilm Life Science, Licor Biosciences, UVP and Syngene.

Purification: This product was aseptically filtered through a Millipore 0.22 micron filter into clean, pre-sterilized containers. The product was tested on trypticase soy agar for 24 hours, 48 hours and 72 hours and was found to be negative for bacteria.

Sterility: Sterile filtered

Application Details

Application Notes: Western Blot : User Defined
Other Dilution: Optimum performance is achieved using this product undiluted. However, this blocking solution can often be diluted 1:1 in TBS without a significant loss of performance.

Comment:

Synonyms: Multiplex Blocking Buffer, Fluorescent Blocking Buffer, Blocking Solution, Blocking Buffer Western Blot, IRDye Western Blot Blocking Buffer, Alexa Dye Blocking Buffer, DyLight Blocking Buffer

Protocol: Fluorescence technology is widely used to detect proteins in both the visible and near-infrared ranges. This product allows for superior signal detection and lower background noise when fluorochrome conjugated antibodies are used to visualize proteins in western blotting and other applications. Antibody conjugates prepared with IRDye® 800 and IRDye® 700DX (Licor), Cy2™, Cy3™, Cy3.5™, Cy5™ and Cy5.5™ (GE Healthcare), DyLight™405, DyLight™ 549, DyLight™ 649, DyLight™ 680, and DyLight™ 800 (Thermo Fisher/Pierce) and Alexa Fluor® 488, Alexa Fluor® 532, Alexa Fluor® 546, Alexa Fluor® 647 and Alexa Fluor® 680 (Invitrogen/Molecular Probes) have been validated on various platforms using this product with superior results compared to other commercially available products. In the infrared range, where little to no autofluorescence occurs, specific signal is sharply evident from any background giving the best possible signal-to-noise ratio. This allows for detection levels in the picogram range which rivals the sensitivity of chemiluminescence on film for western blotting. Superior results are also seen when this product is used for simultaneous labeling (multiplex) in western blots or microscopy using various fluorochrome combinations for multicolor imaging. Membranes blocked with this product can be dried and are very stable. Membranes that are stored protected from light can be re-washed and/or rescanned.

Restrictions: For Research Use only

Handling

Format: Liquid

Concentration: 1 X

Preservative: Thimerosal (Merthiolate)

Precaution of Use: This product contains thimerosal (merthiolate): a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: 4 °C

Storage Comment: Store Blocking Buffer at 4° C prior to opening. DO NOT FREEZE.

References

Spence, Dube, Uezu, Locke, Soderblom, Soderling: "In vivo proximity proteomics of nascent synapses reveals a novel regulator of cytoskeleton-mediated synaptic maturation." in: Nature communications, Vol. 10, Issue 1, pp. 386, (2019) (PubMed).

Fühner, Heine, Helmsing, Goy, Heidepriem, Loeffler, Dübel, Gerhard, Hust: "Development of Neutralizing and Non-neutralizing Antibodies Targeting Known and Novel Epitopes of TcdB of Clostridioides difficile." in: Frontiers in microbiology, Vol. 9, pp. 2908, (2018) (PubMed).

Smith, Saul, Barton, Luna: "Generation and characterization of monoclonal antibodies that recognize human and murine supervillin protein isoforms." in: PLoS ONE, Vol. 13, Issue 10, pp. e0205910, (2018) (PubMed).

Pereira, Bacman, Arguello, Zekonyte, Williams, Edgell, Moraes: "mitoTev-TALE: a monomeric DNA editing enzyme to reduce mutant mitochondrial DNA levels." in: EMBO molecular medicine, Vol. 10, Issue 9, (2018) (PubMed).

Yang, Mott, Sutiwisesak, Lu, Raso, Stowell, Babunovic, Lee, Carpenter, Way, Fortune, Behar: "Mycobacterium tuberculosis-specific CD4+ and CD8+ T cells differ in their capacity to recognize infected macrophages." in: PLoS pathogens, Vol. 14, Issue 5, pp. e1007060, (2018) (PubMed).

Jayaram, Runa, Kemp, Payne: "Nanoparticle-induced oxidation of corona proteins initiates an oxidative stress response in cells." in: Nanoscale, Vol. 9, Issue 22, pp. 7595-7601, (2018) (PubMed).

Loeffler, Foertsch, Popov, Mattes, Schlageter, Sedlmayr, Ridder, Dang, von Bojničić-Kninski, Weber, Fischer, Greifenstein, Bykovskaya, Buliev, Bischoff, Hahn, Meier, Bräse, Powell, Balaban, Breitling et al.: "High-flexibility combinatorial peptide synthesis with laser-based transfer of monomers in solid matrix material. ..." in: Nature communications, Vol. 7, pp. 11844, (2018) (PubMed).

Freire, Pol-Fachin, Coêlho, Viana, Magalhães, Cordeiro, Fischer, Loeffler, Jaenisch, Franca, Marques, Lins: "Mapping Putative B-Cell Zika Virus NS1 Epitopes Provides Molecular Basis for Anti-NS1 Antibody Discrimination between Zika and Dengue Viruses." in: ACS omega, Vol. 2, Issue 7, pp. 3913-3920, (2017) (PubMed).

Klein, Takahashi, Ma, Stagi, Zhou, Lam, Strittmatter: "Loss of TMEM106B Ameliorates Lysosomal and Frontotemporal Dementia-Related Phenotypes in Progranulin-Deficient Mice." in: Neuron, Vol. 95, Issue 2, pp. 281-296.e6, (2017) (PubMed).

Davey, Riley, Andrews, Tyminski, Limberis, Pogoriler, Partridge, Olive, Hedrick, Flake, Peranteau: "Induction of Immune Tolerance to Foreign Protein via Adeno-Associated Viral Vector Gene Transfer in Mid-Gestation Fetal Sheep." in: PLoS ONE, Vol. 12, Issue 1, pp. e0171132, (2017) (PubMed).

Takahashi, Klein, Bhagat, Kaufman, Kostylev, Ikezu, Strittmatter: "Opposing effects of progranulin deficiency on amyloid and tau pathologies via microglial TYROBP network." in: Acta neuropathologica, Vol. 133, Issue 5, pp. 785-807, (2017) (PubMed).

Morales Betanzos, Federspiel, Palubinsky, McLaughlin, Liebler: "Dynamic Phosphorylation of Apoptosis Signal Regulating Kinase 1 (ASK1) in Response to Oxidative and Electrophilic Stress." in: Chemical research in toxicology, Vol. 29, Issue 12, pp. 2175-2183, (2017) (PubMed).

Wang, Wang, Xu, Biao, Yang: "A single amino acid substitution alter antigenicity of Glycosylated protein 4 of HP-PRRSV." in: Virology journal, Vol. 13, Issue 1, pp. 129, (2017) (PubMed).

Maccari, Genoni, Sansonno, Toniolo: "Properties of Two Enterovirus Antibodies that are Utilized in Diabetes Research." in: Scientific reports, Vol. 6, pp. 24757, (2017) (PubMed).

Hu, Azeloglu, Ron, Tran-Ba, Calizo, Tavassoly, Bhattacharya, Jayaraman, Chen, Rabinovich, Iyengar, Hone, He, Kaufman: "A biomimetic gelatin-based platform elicits a pro-differentiation effect on podocytes through mechanotransduction." in: Scientific reports, Vol. 7, pp. 43934, (2017) (PubMed).

Kessler, Laggai, Van Wonterg, Gemperlein, Müller, Haybaeck, Vandenbroucke, Ogris, Libert, Kiemer: "Transient Hepatic Overexpression of Insulin-Like Growth Factor 2 Induces Free Cholesterol and Lipid Droplet Formation." in: Frontiers in physiology, Vol. 7, pp. 147, (2016) (PubMed).

Sun, Wang, Wen, Bao, Shi, Tao, Jiang, Zeng, Xu, Tian, Zheng, Chen: "Identification of a Highly Conserved Epitope on Avian Influenza Virus Non-Structural Protein 1 Using a Peptide Microarray." in: PLoS ONE, Vol. 11, Issue 3, pp. e0149868, (2016) (PubMed).

Metz, Beißbarth, Ellenberger, Pache, Stork, Ringelstein, Aktas, Jarius, Wildemann, Dihazi, Friede, Brück, Ruprecht, Paul: "Serum peptide reactivities may distinguish neuromyelitis optica subgroups and multiple sclerosis." in: Neurology(R) neuroimmunology & neuroinflammation, Vol. 3, Issue 2, pp. e204, (2016) (PubMed).

Simpson-Lavy, Bronstein, Kupiec, Johnston: "Cross-Talk between Carbon Metabolism and the DNA Damage Response in S. cerevisiae." in: Cell reports, Vol. 12, Issue 11, pp. 1865-75, (2016) (PubMed).

Mock, Warta, Geisenberger, Bischoff, Schulte, Lamszus, Stadler, Felgenhauer, Schichor, Schwartz, Matschke, Jungk, Ahmadi, Sahm, Capper, Glass, Tonn, Westphal, von Deimling, Unterberg, Bermejo et al.: "Printed peptide arrays identify prognostic TNC serumantibodies in glioblastoma patients. ..." in: Oncotarget, Vol. 6, Issue 15, pp. 13579-90, (2016) (PubMed).