NEW SUMO1 Antibody Mouse Monoclonal (Clone 5D8B16)

SUMO1 Antibody Mouse Monoclonal (Clone 5D8B16)
$0.00

Host/Isotype
Mouse / IgG2b

Clone
5D8B16

Species Reactivity
Broad Reactivity

Validation Data
see datasheet

Anti-SUMO1 antibody, ASM11, is a SUMO1 mouse monoclonal antibody that is part of the Signal-Seeker™ product line. Anti-SUMO1 mouse monoclonal antibody was raised against full-length recombinant SUMO1 protein (Uniprot: P63165). The antibody has been shown to recognize a broad profile of SUMO1 proteins in HAP1, HeLa, and 293 cell lysates (Fig. 1A) and to detect sub-nanogram amounts of recombinant SUMO1 (Fig. 1B). The antibody shows high specificity (Fig. 1A  WT vs KO lane).with no cross reactivity to SUMO2 (Fig. 1B). Furthermore, the sensitivity of  ASM01 was compared to commercially available SUMO1 antibodies, and the data shows that ASM01 detects a more robust and specific profile of SUMO1 modified proteins  (Fig. 1C). ASM01 is purified by Protein G affinity chromatography and is supplied as a lyophilized white powder. 

Each Lot of antibody is quality controlled to provide a high batch to batch consistency. The Lot specific µg per tube can be found in the Lot specific COA documents.


Figure 1: Western Blot using SUMO-2/3 Antibody

ASM01 was used at a 1:5000 dilution (0.2 µg/mL) following the recommended western blot protocol (see above). Figure 1A: HAP1 wildtype (WT) or SUMO1 knockout (KO) lysate, HeLa cell +/- NEM lysate and 293 cell +/- NEM lysate was obtained using BlastR lysis and filter system.  10 µg of each lysate were separated by SDS-PAGE and transferred to PVDF. Robust SUMO1 profiles were detected for every cell type. Specificity is shown by the lack of SUMO1 detection in SUMO1 KO cells and significantly diminished profiles in lysates prepared in the absence of the SENP inhibitor (NEM).

Figure 1B: Titration of recombinant SUMO1 lanes 1-6 contain 5.0, 2.5, 1.25, 0.6, 0.3, and 0.15 ng SUMO1, while lane 7 contains 1000 ng of recombinant SUMO2. 

Figure 1C: HAP1 WT and KO lysate as prepared in 1A was used. 10 µg of each lysate were separated by SDS-PAGE and transferred to PVDF. SUMO1 proteins were detected using the recommended concentrations of ASM01 (Cytoskeleton), 21C7 (Invitrogen—purified), 21C7 (DSHB—supernatant), and D11 (Santa Cruz) as shown in the figure. All samples were developed and imaged simultaneously to ensure identical experimental conditions. To see the full Western blot protocol, see the product datasheet.

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Immunofluorescence applications for ASM01 have not been tested

    For more information contact:  signalseeker@cytoskeleton.com

    Associated Products:

    Signal-Seeker™ SUMOylation 1 Detection Kit (Cat. # BK165)

    Signal-Seeker™ SUMOylation 1 Affinity Beads (Cat.# ASM11-beads)

    Signal-Seeker™: BlastR™ Rapid Lysate Prep Kit (Cat. # BLR01)

    For product Datasheets and MSDSs please click on the PDF links below.

    Sample Size Datasheet (Cat. ASM01-S):  

    Certificate of Analysis:  available upon request

     

    New product released in 2018!   For the most recent publications citing this and other Signal-Seeker™ products, see our Signal-Seeker™ Validation Data Page click here

    Visit our Signal-Seeker™ Tech Tips and FAQs page for technical tips and frequently asked questions regarding this and other Signal-Seeker™ products click here

     

    If you have any questions concerning this product, please contact our Technical Service department at tservice@cytoskeleton.com