Description:

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.

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