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SUMO I Antibody

(Catalog # AP018:100ul)

 

Specificity: Rabbit polyclonal antibody against Human SUMO I (Ubiquitin-like protein SMT3C).

 

Antigen used for immunizations: Prokaryotic recombinant protein corresponding to the full expressed region of the human Ubiquitin-like protein SMT3C precursor [access no: Q93068, also called Ubiquitin-homology domain protein PIC1, Ubiquitin-like protein UBL1, Ubiquitin-related protein SUMO-1, GAP modifying protein 1, GMP1, and Sentrin].

 

Storage and Reconstitution: The  antiserum was stored in lyophilized form.  Before it is used, it should be reconstitited with 100 ul of H2O.  Working dilution on Western blotting is 1:1000.  

 

Recommendations on use:

Immunofluorescence:

Not applicable.

Quality of anti-SUMO I antibody

The quality of anti-SUMO I antibody at 1,000-fold dilution is good to detect SUMO I at the concentration as low as 5 ng of SUMO I. SUMO I was run on a 15% SDS-PAGE, followed by transferring to PVDF membrane and immunoblotting with 1,000-fold diluted.
Lane 1, protein marker
Lane 2, 5 ng;   Lane 3, 10 ng;   Lane 4, 25 ng;   Lane 5, 50 ng;
Lane 6, 100 ng;   Lane 7, 200 ng of SUMO I.



Western blots (12.5% SDS-PAGE) of sumoylated 50Kd-Topo I substrate with anti-topo I antibody (AP006) and anti-SUMO I antibody (AP018)

Lane 1 & 2 are blotted with 1,000-fold diluted anti-topo I antibody (AP006).
Lane 3 & 4 are blotted with 1,000-fold diluted anti-SUMO I antibody (AP018).
Lane 1 and Lane 3 are 50Kd-Topo I substrate (B007).
Lane 2 and Lane 4 are sumoylated 50Kd-Topo I substrate which are obtained by incubating 2 ml of 50Kd-Topo I substrate with 2 ml E1 (P001), 2 ml of E2 (P002), 2 ml of SUMO I (P003), 2 ml of 10X sumoylation buffer (B006), 2 ml of 20 mM ATP and 8 ml of H2O at 37oC for 45 min.

 

Storage and stability: Store unopened antibody at 4oC or lower. Under these conditions, there is no significant loss in product performance up to one year from the purchase date. The reconstituted antibody is stable for at least two months when stored at 4oC.  For long term storage, it is recommended that aliquots of the antibody are frozen at -20oC (frost-free freezers are not recommended). Repeated freezing and thawing must be avoided. Prepare working dilutions on the day of use.

 

Legal consideration: FOR RESEARCH USE ONLY.