Overview

  • Product name

    Anti-SENP1 antibody [EPR3844]
    See all SENP1 primary antibodies
  • Description

    Rabbit monoclonal [EPR3844] to SENP1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt, ICC/IF, WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human SENP1 aa 400-500. The exact sequence is proprietary.
    Database link: Q9P0U3

  • Positive control

    • WB: HeLa, HUVEC, Jurkat, Daudi and U87-MG cell lysates. IHC-P: Human testis tissue. ICC/IF: Jurkat cells. Flow Cyt: HeLa cells.
  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab108981 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt 1/20.
ICC/IF 1/100 - 1/500.
WB 1/1000 - 1/20000. Predicted molecular weight: 73 kDa.
IHC-P 1/100 - 1/300. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Target

  • Function

    Protease that catalyzes two essential functions in the SUMO pathway: processing of full-length SUMO1, SUMO2 and SUMO3 to their mature forms and deconjugation of SUMO1, SUMO2 and SUMO3 from targeted proteins. Deconjugates SUMO1 from HIPK2. Deconjugates SUMO1 from HDAC1, which decreases its transcriptional repression activity.
  • Tissue specificity

    Highly expressed in testis. Expressed at lower levels in thymus, pancreas, spleen, liver, ovary and small intestine.
  • Sequence similarities

    Belongs to the peptidase C48 family.
  • Cellular localization

    Nucleus. Cytoplasm. Shuttles between cytoplasm and nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • SENP 1 antibody
    • SENP1 antibody
    • SENP1_HUMAN antibody
    • Sentrin specific protease 1 antibody
    • Sentrin-specific protease 1 antibody
    • Sentrin/SUMO specific protease 1 antibody
    • Sentrin/SUMO specific protease antibody
    • Sentrin/SUMO specific protease SENP 1 antibody
    • Sentrin/SUMO specific protease SENP1 antibody
    • Sentrin/SUMO-specific protease SENP1 antibody
    • SUMO1/sentrin specific peptidase 1 antibody
    • SUMO1/sentrin specific protease 1 antibody
    • SuPr 2 antibody
    • SuPr2 antibody
    see all

Images

  • Anti-SENP1 antibody [EPR3844] (ab108981) at 1/20000 dilution (purified) + Daudi cell lysate at 10 µg

    Secondary
    HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution

    Predicted band size: 73 kDa
    Observed band size: 73 kDa



    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • Immunofluorescence staining of Jurkat cells with purified ab108981 at a working dilution of 1/100, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab108981 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

  • Immunohistochemical staining of paraffin embedded human testis with purified ab108981 at a working dilution of 1/300. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
  • Flow Cytometry analysis of HeLa cells labelling SENP1 with purified ab108981 at a dilution of 1/20 (red). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. An Alexa Flour® 488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • Anti-SENP1 antibody [EPR3844] (ab108981) at 1/20000 dilution (purified) + U87-MG cell lysate at 10 µg

    Secondary
    HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution

    Predicted band size: 73 kDa
    Observed band size: 73 kDa



    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • Unpurified ab108981 (1/500) staining SENP1 in HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.5% Triton X-100/PBS and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details please refer to Abreview.

    See Abreview

  • All lanes : Anti-SENP1 antibody [EPR3844] (ab108981) at 1/1000 dilution (unpurified)

    Lane 1 : HeLa cell lysates
    Lane 2 : HUVEC cell lysates
    Lane 3 : Jurkat cell lysates
    Lane 4 : Daudi cell lysates
    Lane 5 : U87-MG cell lysates

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Standard HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 73 kDa

  • All lanes : Anti-SENP1 antibody [EPR3844] (ab108981) at 1/1000 dilution (unpurified)

    Lane 1 : COS-1 cell lysate
    Lane 2 : COS-1 cell lysate transfected with SENP1
    Lane 3 : COS-1 cell lysate transfected with SENP1 mutant

    Lysates/proteins at 20000 cells per lane.

    Secondary
    All lanes : HRP-conjugated donkey anti-rabbit IgG polyclonal at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 73 kDa
    Observed band size: 75 kDa
    why is the actual band size different from the predicted?


    Exposure time: 6 minutes


    Blocked with 3% milk for 1 hour at 25°C.

    Incubated with the primary antibody for 18 hours at 4°C.

    See Abreview

References

This product has been referenced in:

  • Hsu HL  et al. Chloramphenicol Induces Autophagy and Inhibits the Hypoxia Inducible Factor-1 Alpha Pathway in Non-Small Cell Lung Cancer Cells. Int J Mol Sci 20:N/A (2019). Read more (PubMed: 30609861) »
  • Liebelt F  et al. SUMOylation and the HSF1-Regulated Chaperone Network Converge to Promote Proteostasis in Response to Heat Shock. Cell Rep 26:236-249.e4 (2019). Read more (PubMed: 30605679) »
See all 18 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Application
Western blot
Sample
Mouse Cell lysate - nuclear (MEF)
Gel Running Conditions
Reduced Denaturing (8)
Loading amount
20 µg
Specification
MEF
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Mar 07 2019

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Permeabilization
Yes - 0.5% Triton 5 minutes
Specification
HeLa
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Oct 20 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa cells)
Gel Running Conditions
Reduced Denaturing (handcast 10% tris-glycine, 15 well)
Loading amount
40000 cells
Treatment
SENP1 siRNA treated to show specificity
Specification
HeLa cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Jun 14 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa)
Gel Running Conditions
Reduced Denaturing
Loading amount
10 µg
Specification
HeLa
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Feb 24 2017

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa cells)
Permeabilization
Yes - 0.5% TritonX100
Specification
HeLa cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Nov 30 2016

Application
Western blot
Sample
Human Recombinant protein (Human SENP1 transfected into monkey COS-1 cells)
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris (BioRad))
Loading amount
20000 cells
Specification
Human SENP1 transfected into monkey COS-1 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Dr. Ragnhild Eskeland

Verified customer

Submitted Apr 22 2015

Answer

We do not recommend ab108981 for mouse based on the fact that this antibody did not produce any signal on our mouse samples in western blot. We tested mouse heart, brain, kidney and spleen.

Read More
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% Triton-X100 in PBS

Dr. Kirk Mcmanus

Verified customer

Submitted Apr 20 2012

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