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Mono- and polyubiquitinylated conjugates monoclonal antibody (FK2) (HRP conjugate)

BML-PW0150-0025 25 µg 125.00 USD
BML-PW0150-0100 100 µg 420.00 USD
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  • Detects K29-, K48-, and K63-linked mono- and polyubiquitinylated proteins
  • Most cited multi-ubiquitin antibody with over 500 citations
  • Validated for WB, IP, IHC, and ELISA applications
FK2 antibody was conjugated to horseradish peroxidase (HRP) by a proprietary method. The HRP-linked FK2 monoclonal antibody (FK2H™) gives enhanced Western blotting detection of mono- and poly-ubiquitinylated protein conjugates and multi-ubiquitin chains compared to the unconjugated FK2 antibody (Prod. No. BML-PW8810). Shows no reactivity with free ubiquitin. Recognizes K29-, K48-, and K63-linked polyubiquitinylated and monoubiquitinylated proteins but not free ubiquitin.

By using the antibodies in concert the degree of protein ubiquitinylation may be determined. These antibodies have been used to not only investigate the dynamics of ubiquitin conjugation but also for the development of immunoassays permitting quantification of serum and intracellular multiubiquitin chains. The hybridoma secreting the antibody FK2 was generated by fusion of splenocytes from Balb/c mice which had received repeated immunization with a crude preparation of polyubiquitinylated-lysozyme. The antibody (clone FK2) has been extensively characterized by one-dimensional Western blotting. The immunoglobulin subclass is IgG1.

Product Specification

Immunogen:Crude preparation of polyubiquitinylated-lysozyme.
UniProt ID:P0CG47 (UBB), P0CG48 (UBC)
Source:Purified from ascites.
Species reactivity:Species independent
Specificity:Recognizes mono- and polyubiquitinylated protein conjugates in a wide range of species.
Crossreactivity:Does not cross-react with free ubiquitin.
Applications:ELISA, IHC, WB
Recommended Dilutions/Conditions:Western Blot (1:1,000)
Suggested dilutions/conditions may not be available for all applications.
Optimal conditions must be determined individually for each application.
Formulation:Liquid. In PBS.
Use/Stability:Antibody is supplied at a concentration of 1mg/mL. Store at 4ºC. Can be stored at -20ºC if required, but avoid multiple freeze/thawing. Vial contains purified HRP-linked immunoglobulin suspended in phosphate-buffered saline.
Handling:Avoid freeze/thaw cycles.
Shipping:Blue Ice Not Frozen
Long Term Storage:+4°C
Technical Info/Product Notes:FK2 antibody was conjugated to horseradish peroxidase (HRP) by a proprietary method. The HRP-linked FK2 monoclonal antibody (FK2H) gives enhanced western blotting detection of mono- and poly-ubiquitinylated protein conjugates and multi-ubiquitin chains compared to the unconjugated FK2 antibody (PW8810). For western blotting an initial antibody dilution of 1:1000 is recommended, followed by washing and signal detection using appropriate development reagents.
Note: Milk should NOT be used in blocking/antibody binding solutions. 1% BSA in PBS or TBS Tween should be used instead.
Regulatory Status:RUO - Research Use Only
Mono- and polyubiquitinylated conjugates monoclonal antibody (FK2) (HRP conjugate) WB
Figure: Lanes a-d, HeLa S3 S100 cytosol preparation (SW8750; 10µg per lane); lanes e-h, K48-linked multiubiquitin chains (Prod. No: BML-PW8860; 0.5µg per lane). Lanes a & e probed with Prod. No BML-PW8805 (clone FK1; 1µg/mL); lanes b & f probed with Prod. No. BML-PW0145 (FK1H™; 1µg/mL); lanes c & g probed with Prod. No. BML-PW8810 (clone FK2; 1µg/mL); lanes d & h probed with Prod. No. BML-PW0150 (FK2H™; 1µg/mL).
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Mono- and polyubiquitinylated conjugates monoclonal antibody (FK2) (HRP conjugate) WB

Product Literature References

Age-dependent degeneration of an identified adult leg motor neuron in a Drosophila SOD1 model of ALS: A. Agudelo, et al.; Biol. Open (2020), Abstract;
Knockdown of MSI2 inhibits metastasis by interacting with caveolin-1 and inhibiting its ubiquitylation in human NF1-MPNST cells: K. Yang, et al.; Cell Death Dis. 11, 489 (2020), Abstract; Full Text
Absence of Cytosolic 2-Cys Prx Subtypes I and II Exacerbates TNF-α-Induced Apoptosis via Different Routes: S. Lee, et al.; Cell Rep. 26, 2194 (2019), Abstract;
Honokiol Enhances TRAIL-Mediated Apoptosis through STAMBPL1-Induced Survivin and c-FLIP Degradation: S.M. Woo, et al.; Biomolecules 6, 9 (2019), Abstract; Full Text
The Deubiquitinating Enzyme Inhibitor PR-619 Enhances the Cytotoxicity of Cisplatin via the Suppression of Anti-Apoptotic Bcl-2 Protein: In Vitro and In Vivo Study: KL. Kuo, et al.; Cells 8, 1268 (2019), Abstract; Full Text
ABIN1 inhibits HDAC1 ubiquitination and protects it from both proteasome- and lysozyme-dependent degradation: Y. Ma, et al.; J. Cell. Biochem. 119, 3030 (2018), Abstract;
Pancreatic β cells overexpressing hIAPP impaired mitophagy and unbalanced mitochondrial dynamics: M.G. Hernandez, et al.; Cell Death Dis. 9, 481 (2018), Abstract; Full Text
USP7 and USP47 deubiquitinases regulate NLRP3 inflammasome activation: P. Palazon-Riquelme, et al.; EMBO Rep. 19, e44766 (2018), Abstract; Full Text
High Glucose Stimulates Mineralocorticoid Receptor Transcriptional Activity Through the Protein Kinase C β Signaling: T. Hayashi, et al.; Int. Heart J. 58, 794 (2017), Abstract;
Selective degradation of PU.1 during autophagy represses the differentiation and antitumour activity of TH9 cells: T. Rivera-Vargas, et al.; Nat. Commun. 8, 559 (2017), Abstract; Full Text
TSC2 N-terminal lysine acetylation status affects to its stability modulating mTORC1 signaling and autophagy: A. García-Aguilar , et al.; Biochem. Biophys. Acta 1863, 2658 (2016), Application(s): Western blot, Abstract;
Efficient APC/C substrate degradation in cells undergoing mitotic exit depends on K11 ubiquitin linkages: M. Min, et al.; Mol. Biol. Cell 26, 4325 (2015), Application(s): Western Blot, Abstract;
PI3K/AKT activation induces PTEN ubiquitination and destabilization accelerating tumourigenesis: M.S. Lee, et al.; Nat. Commun. 6, 7769 (2015), Application(s): Western Blot, Abstract; Full Text
Severe acute respiratory syndrome coronavirus protein 6 mediates ubiquitin-dependent proteosomal degradation of N-Myc (and STAT) interactor: W. Cheng, et al.; Virol Sin. 30, 153 (2015), Abstract;
Hectd1 regulates intracellular localization and secretion of Hsp90 to control cellular behavior of the cranial mesenchyme: A. Sarkar & I. Zohn; J. Cell Biol. 196, 789 (2012), Application(s): IF, WB of mouse embryonic tissue, Abstract; Full Text
Selective degradation of aggregate-prone CryAB mutants by HSPB1 is mediated by ubiquitin-proteasome pathways: H. Zhang, et al.; J. Mol. Cell. Cardiol. 49, 918 (2010), Application(s): WB of embryonic rat cardiac cells, Abstract; Full Text

General Literature References

Dynamics of ubiquitin conjugation during heat-shock response revealed by using a monoclonal antibody specific to multi-ubiquitin chains: M. Fujimuro et al.; Eur. J. Biochem. 249, 427 (1997), Abstract;
Serum concentrations of free ubiquitin and multiubiquitin chains: K. Takada et al.; Clin. Chem. 43, 1188 (1997), Abstract;
Immunoassay for the quantification of intracellular multi-ubiquitin chains: K. Takada et al.; Eur. J. Biochem. 233, 42 (1995), Abstract;
Production and characterization of monoclonal antibodies specific to multi-ubiquitin chains of polyubiquitinated proteins: M. Fujimuro et al.; FEBS Lett. 349, 173 (1994), Abstract;

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