Description:

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:

Clone:	FK2
Host:	Mouse
Isotype:	IgG1
Immunogen:	Crude preparation of polyubiquitinylated-lysozyme.
UniProt ID:	P0CG47 (UBB), P0CG48 (UBC)
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.

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