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RecombinantRabMAb

Recombinant Anti-RanGAP1 antibody [EPR3295] (ab92360)

  • Datasheet
Reviews (3) Submit a question References (8)

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Western blot - Anti-RanGAP1 antibody [EPR3295] (ab92360)
  • Western blot - Anti-RanGAP1 antibody [EPR3295] (ab92360)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RanGAP1 antibody [EPR3295] (ab92360)
  • Immunocytochemistry/ Immunofluorescence - Anti-RanGAP1 antibody [EPR3295] (ab92360)
  • Flow Cytometry - Anti-RanGAP1 antibody [EPR3295] (ab92360)
  • Immunoprecipitation - Anti-RanGAP1 antibody [EPR3295] (ab92360)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RanGAP1 antibody [EPR3295] (ab92360)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RanGAP1 antibody [EPR3295] (ab92360)
  • Immunocytochemistry/ Immunofluorescence - Anti-RanGAP1 antibody [EPR3295] (ab92360)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RanGAP1 antibody [EPR3295] (ab92360)
  • Immunocytochemistry/ Immunofluorescence - Anti-RanGAP1 antibody [EPR3295] (ab92360)
  • Flow Cytometry - Anti-RanGAP1 antibody [EPR3295] (ab92360)
  • Anti-RanGAP1 antibody [EPR3295] (ab92360)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3295] to RanGAP1
  • Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IF
  • Reacts with: Mouse, Rat, Human

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Overview

  • Product name

    Anti-RanGAP1 antibody [EPR3295]
    See all RanGAP1 primary antibodies
  • Description

    Rabbit monoclonal [EPR3295] to RanGAP1
  • Host species

    Rabbit
  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Mouse
    Rat
    Human
    IP
    Human
    WB
    Mouse
    Rat
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide. within Human RanGAP1 aa 1-100. The exact sequence is proprietary.
    Database link: P46060

  • Positive control

    • WB: MCF7 (Human breast adenocarcinoma cell line) Cytoplasmic Lysate - tumor cell line (ab29538), HeLa, SH SY5Y and A549 cell lysates, mouse and rat brain tissue lysates. IHC-P: Human breast carcinoma, Human testis, Human bladder carcinoma, Mouse testis and Rat liver tissue. ICC/IF: mouse hepatocyte, and MCF7 cells. Flow Cyt: Jurkat cells IP: HeLa cell lysate
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR3295
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Cell Cycle
    • Cell Division
    • Centromere
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • Nuclear Receptors
    • Nuclear Pore Complex

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-RanGAP1 antibody [EPR3295] (ab204650)
    • Alexa Fluor® 647 Anti-RanGAP1 antibody [EPR3295] (ab204651)
    • HRP Anti-RanGAP1 antibody [EPR3295] (ab205449)
    • Anti-RanGAP1 antibody [EPR3295] - BSA and Azide free (ab239907)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human RanGAP1 protein (ab131750)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab92360 in the following tested applications.

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

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
Flow Cyt
Human
ICC/IF
Human
IHC-P
Mouse
Rat
Human
IP
Human
WB
Mouse
Rat
Human
Application Abreviews Notes
WB (1)
1/1000 - 1/5000. Predicted molecular weight: 64 kDa.
IP
1/20.

For unpurified use at 1/10

IHC-P
1/500. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

For unpurified use at 1/100 - 1/250.

See IHC antigen retrieval protocols.

Flow Cyt (1)
1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

For unpurified use at 1/50

ICC/IF (1)
1/100 - 1/250.
Notes
WB
1/1000 - 1/5000. Predicted molecular weight: 64 kDa.
IP
1/20.

For unpurified use at 1/10

IHC-P
1/500. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

For unpurified use at 1/100 - 1/250.

See IHC antigen retrieval protocols.

Flow Cyt
1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

For unpurified use at 1/50

ICC/IF
1/100 - 1/250.

Target

  • Function

    GTPase activator for the nuclear Ras-related regulatory protein Ran, converting it to the putatively inactive GDP-bound state.
  • Tissue specificity

    Highly expressed in brain, thymus and testis.
  • Sequence similarities

    Belongs to the RNA1 family.
    Contains 6 LRR (leucine-rich) repeats.
  • Post-translational
    modifications

    Phosphorylated occurs before nuclear envelope breakdown and continues throughout mitosis. Phosphorylated by the M-phase kinase cyclin B/Cdk1, in vitro. Differential timimg of dephosphorylation occurs during phases of mitosis. The phosphorylated form remains associated with RANBP2/NUP358 and the SUMO E2-conjugating enzyme, UBC9, on nuclear pore complex (NPC) diassembly and during mitosis.
    Sumoylated with SUMO1. Sumoylation is necessary for targeting to the nuclear envelope (NE), and for association with mitotic spindles and kinetochores during mitosis. Also required for interaction with RANBP2 and is mediated by UBC9.
  • Cellular localization

    Cytoplasm. Nucleus membrane. Chromosome, centromere, kinetochore. Cytoplasm, cytoskeleton, spindle pole. Cytoplasmic during interphase. Targeted to the nuclear rim after sumoylation. During mitosis, associates with mitotic spindles. Association with kinetochores appears soon after nuclear envelope breakdown and persists until late anaphase. Mitotic location also requires sumoylation.
  • Target information above from: UniProt accession P46060 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 5905 Human
    • Entrez Gene: 19387 Mouse
    • Entrez Gene: 362965 Rat
    • Omim: 602362 Human
    • SwissProt: P46060 Human
    • SwissProt: P46061 Mouse
    • Unigene: 183800 Human
    • Unigene: 270975 Mouse
    • Unigene: 475271 Mouse
    • Unigene: 107099 Rat
    see all
  • Alternative names

    • Fug 1 antibody
    • Fug1 antibody
    • GTPase-activating protein, RAN, 1 antibody
    • KIAA1835 antibody
    • MGC20266 antibody
    • OTTHUMP00000028918 antibody
    • OTTHUMP00000198755 antibody
    • OTTHUMP00000198756 antibody
    • OTTHUMP00000198757 antibody
    • OTTHUMP00000198758 antibody
    • RAGP1_HUMAN antibody
    • Ran 1 antibody
    • RAN GTPase activating protein 1 antibody
    • Ran GTPase-activating protein 1 antibody
    • Ran1 antibody
    • RANGAP 1 antibody
    • RANGAP antibody
    • RanGAP1 antibody
    • SD antibody
    • Segregation distorter homolog antibody
    • Segregation distortion antibody
    see all

Images

  • Western blot - Anti-RanGAP1 antibody [EPR3295] (ab92360)
    Western blot - Anti-RanGAP1 antibody [EPR3295] (ab92360)
    All lanes : Anti-RanGAP1 antibody [EPR3295] (ab92360) at 1/1000 dilution (Purified)

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
    Lane 2 : SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysates
    Lane 3 : Mouse brain lysates
    Lane 4 : Rat brain lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 64 kDa
    Observed band size: 70,90 kDa
    why is the actual band size different from the predicted?



    The doublets detected are consistent with what have been described in literature PMID: 24988324 and PMID: 21646468

  • Western blot - Anti-RanGAP1 antibody [EPR3295] (ab92360)
    Western blot - Anti-RanGAP1 antibody [EPR3295] (ab92360)
    All lanes : Anti-RanGAP1 antibody [EPR3295] (ab92360) at 1/1000 dilution

    Lane 1 : HeLa cell lysate
    Lane 2 : MCF-7 cell lysate
    Lane 3 : SH-SY5Y cell lysate
    Lane 4 : A549 cell lysate

    Lysates/proteins at 10 µg per lane.

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

    Predicted band size: 64 kDa
    Observed band size: 64 kDa
    Additional bands at: 90 kDa. We are unsure as to the identity of these extra bands.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RanGAP1 antibody [EPR3295] (ab92360)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RanGAP1 antibody [EPR3295] (ab92360)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human bladder carcinoma tissue sections labeling RanGAP1 with purified ab92360 at 1/500 dilution (0.22 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • Immunocytochemistry/ Immunofluorescence - Anti-RanGAP1 antibody [EPR3295] (ab92360)
    Immunocytochemistry/ Immunofluorescence - Anti-RanGAP1 antibody [EPR3295] (ab92360)

    Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling RanGAP1 with purified ab92360 at 1/100 dilution (1.1 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 (2.5 µg/ml) dilution. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • Flow Cytometry - Anti-RanGAP1 antibody [EPR3295] (ab92360)
    Flow Cytometry - Anti-RanGAP1 antibody [EPR3295] (ab92360)

    Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling RanGAP1 with purified ab92360 at 1/20 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

  • Immunoprecipitation - Anti-RanGAP1 antibody [EPR3295] (ab92360)
    Immunoprecipitation - Anti-RanGAP1 antibody [EPR3295] (ab92360)

    ab92360 (purified) at 1/20 dilution (0.5ug) immunoprecipitating RanGAP1 in HeLa whole cell lysates.
    Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates 10ug
    Lane 2 (+): ab92360 & HeLa whole cell lysates
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab92360 in HeLa whole cell lysates
    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RanGAP1 antibody [EPR3295] (ab92360)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RanGAP1 antibody [EPR3295] (ab92360)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat liver tissue sections labeling RanGAP1 with purified ab92360 at 1/500 dilution (0.22 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RanGAP1 antibody [EPR3295] (ab92360)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RanGAP1 antibody [EPR3295] (ab92360)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse testis tissue sections labeling RanGAP1 with purified ab92360 at 1/500 dilution (0.22 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • Immunocytochemistry/ Immunofluorescence - Anti-RanGAP1 antibody [EPR3295] (ab92360)
    Immunocytochemistry/ Immunofluorescence - Anti-RanGAP1 antibody [EPR3295] (ab92360)

    Immunofluorescence staining of MCF7 cells with purified ab92360 at a working dilution of 1/500, counter-stained with DAPI. The secondary antibody was an Alexa Fluor® 488 conjugated goat anti-rabbit (ab150077), used at a dilution of 1/1000. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative control is shown in bottom right hand panel - for the negative control, PBS was used instead of the primary antibody.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RanGAP1 antibody [EPR3295] (ab92360)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RanGAP1 antibody [EPR3295] (ab92360)

    b92360 at 1/100 dilution staining RanGAP1 in paraffin-embedded (1) Human breast carcinoma tissue and (2) Human testis tissue by immunohistochemistry.

    Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

    Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-RanGAP1 antibody [EPR3295] (ab92360)
    Immunocytochemistry/ Immunofluorescence - Anti-RanGAP1 antibody [EPR3295] (ab92360)This image is courtesy of an anonymous Abreview

    ab92360 staining RanGAP1 in mouse hepatocyte cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized, and blocked with 2% BSA for 2 hours at 22°C. Samples were incubated with primary antibody (1/100 in blocking buffer) for 18 hours at 4°C. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal (1/10000) was used as the secondary antibody.

    See Abreview

  • Flow Cytometry - Anti-RanGAP1 antibody [EPR3295] (ab92360)
    Flow Cytometry - Anti-RanGAP1 antibody [EPR3295] (ab92360)
    Overlay histogram showing Jurkat cells stained with ab92360 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab92360, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line). Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in Jurkat cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • Anti-RanGAP1 antibody [EPR3295] (ab92360)
    Anti-RanGAP1 antibody [EPR3295] (ab92360)

Protocols

  • Flow cytometry protocols
  • Immunoprecipitation protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

    • Datasheet
  • References (8)

    Publishing research using ab92360? Please let us know so that we can cite the reference in this datasheet.

    ab92360 has been referenced in 8 publications.

    • Carmichael RE  et al. Insulin-dependent GLUT4 trafficking is not regulated by protein SUMOylation in L6 myocytes. Sci Rep 9:6477 (2019). PubMed: 31019221
    • Khosravi B  et al. Cytoplasmic poly-GA aggregates impair nuclear import of TDP-43 in C9orf72 ALS/FTLD. Hum Mol Genet 26:790-800 (2017). PubMed: 28040728
    • Schludi MH  et al. Spinal poly-GA inclusions in a C9orf72 mouse model trigger motor deficits and inflammation without neuron loss. Acta Neuropathol 134:241-254 (2017). PubMed: 28409281
    • Ditsworth D  et al. Mutant TDP-43 within motor neurons drives disease onset but not progression in amyotrophic lateral sclerosis. Acta Neuropathol 133:907-922 (2017). PubMed: 28357566
    • Oldrini B  et al. EGFR feedback-inhibition by Ran-binding protein 6 is disrupted in cancer. Nat Commun 8:2035 (2017). PubMed: 29229958
    • Wang F  et al. Hypoxia regulates sumoylation pathways in intervertebral disc cells: implications for hypoxic adaptations. Osteoarthritis Cartilage 24:1113-24 (2016). PubMed: 26826302
    • Dubuissez M  et al. Protein Kinase C-Mediated Phosphorylation of BCL11B at Serine 2 Negatively Regulates Its Interaction with NuRD Complexes during CD4+ T-Cell Activation. Mol Cell Biol 36:1881-98 (2016). PubMed: 27161321
    • Tang Y  et al. Quantitative proteomic analysis of HER2 normal and overexpressing MCF-7 breast cancer cells revealed proteomic changes accompanied with HER2 gene amplification. J Proteomics 91C:200-209 (2013). PubMed: 23851309

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-3 of 3 Abreviews or Q&A

    Flow Cytometry abreview for Anti-RanGAP1 antibody [EPR3295]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Flow Cytometry
    Fixation
    Formaldehyde
    Permeabilization
    Yes - BD Perm/Wash Buffer
    Sample
    Human Cell (PBMC)
    Specification
    PBMC
    Gating Strategy
    FSC-A/SSC-A
    Preparation
    Cell harvesting/tissue preparation method: Ficoll isolation of PBMC from whole blood, spin down, wash in FACS buffer 1x, fix, wash 2x, and stain with primary antibody overnight.
    Sample buffer: Buffer composition at analysis was 0.1% sodium azide with FBS in phosphate buffered solution.
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 03 2014

    Immunocytochemistry/ Immunofluorescence abreview for Anti-RanGAP1 antibody [EPR3295]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (Hepatocyte)
    Specification
    Hepatocyte
    Blocking step
    BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 22°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Jan 23 2014

    Western blot abreview for Anti-RanGAP1 antibody [EPR3295]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    50 µg
    Gel Running Conditions
    Non-reduced Denaturing (8%)
    Sample
    Mouse Cell lysate - whole cell (primary hepatoctyes)
    Specification
    primary hepatoctyes
    Blocking step
    (agent) for 30 minute(s) · Concentration: 2µg/mL · Temperature: 22°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 01 2013

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