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Mono- and polyubiquitinylated conjugates monoclonal antibody (FK2) (fluorescein labeled)

 
BML-PW1210-0025 25 µl 170.00 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
For ubiquitin signaling pathway, selective autophagy, proteasomal degradation and protein aggregation studies using fluorescence microscopy.

Product Specification

Clone:FK2
 
Host:Mouse
 
Isotype:IgG1
 
Immunogen:Polyubiquitinylated-lysozyme.
 
UniProt ID:P0CG47 (UBB), P0CG48 (UBC)
 
Species reactivity:Species independent
 
Specificity:Recognizes polyubiquitinylated (all linkage types) and monoubiquitinylated proteins in a wide range of species.
 
Crossreactivity:Does not cross-react with free ubiquitin.
 
Applications:ICC
 
Recommended Dilutions/Conditions:Immunocytochemistry (1:250)
Suggested dilutions/conditions may not be available for all applications.
Optimal conditions must be determined individually for each application.
 
Purity Detail:Purified.
 
Formulation:Liquid. In PBS containing 0.01% sodium azide.
 
Use/Stability:Stable for at least 1 year after receipt when stored at -20°C. Reagent is light sensitive; store in the dark. The use of high quality ‘antiserum-grade’ plastic or glass vials is recommended.
 
Handling:Protect from light. Avoid freeze/thaw cycles. After opening, prepare aliquots and store at -20°C.
 
Shipping:Blue Ice Not Frozen
 
Short Term Storage:+4°C
 
Long Term Storage:+4°C
 
Technical Info/Product Notes:The antibodies BML-PW8805 (clone FK1) and BML-PW8810 (clone FK2) are specific for ubiquitin-protein conjugates and show no reactivity with free ubiquitin. Clone FK1 recognises only polyubiquitinylated proteins and not monoubiquitinylated proteins or free ubiquitin, whilst clone FK2 recognises both mono- and poly-ubiquitinylated species but not free ubiquitin.

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. Over recent years these antibodies have found very wide application in many areas of investigation and there is an associated supporting bibliography.

The hybridoma secreting the antibody FK2 was generated by fusion of splenocytes from Balb/c mice which had received repeated immunisation with a crude preparation of polyubiquitinylated-lysozyme. The antibody (clone FK2) has been extensively characterized by one-dimensional Western blotting and has been shown to recognise polyubiquitinylated (all linkage types) and monoubiquitinylated proteins but not free ubiquitin.

FK2 antibody BML-PW8810 was conjugated to fluorescein by a proprietary method. The fluorescein-labelled FK2 monoclonal antibody (BML-PW1210) facilitates ready detection of mono- and poly-ubiquitinylated protein conjugates in cells by fluorescence microscopy.
 
BML-PW1210_IF
Detection of ubiquitinylated protein conjugates in HeLa cells by fluorescence microscopy using fluorescein labeled FK2 mAb (BML-PW1210), at 1:250 dilution. HeLa cells were treated with proteasome inhibitor MG-132 (5µM, BML-PI102) for 12 hours prior to analysis.
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BML-PW1210_IF

Product Literature References

A Phosphoinositide-binding cluster incavin2 acts as a molecularsensor for cavin1 degradation: V.A. Tillu, et al.; Mol. Biol. Cell 26, 3561 (2015), Application(s): Immunoblotting, Abstract; Full Text
Increased protein aggregation in Zucker Diabetic Fatty rat brain: identification of key mechanistic targets and the therapeutic application of hydrogen sulfide: F. Talaei, et al.; BMC Cell Biol. 15, 1 (2014), 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; Full Text
Serum concentrations of free ubiquitin and multiubiquitin chains: K. Takada et al.; Clin. Chem. 43, 1188 (1997), Abstract; Full Text
Immunoassay for the quantification of intracellular multiubiquitin chains: K. Takada et al.; Eur. J. Biochem. 233, 42 (1995), Abstract; Full Text
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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