Overview

  • Product nameAnti-RanGAP1 antibody [EPR3295]
    See all RanGAP1 primary antibodies
  • Description
    Rabbit monoclonal [EPR3295] to RanGAP1
  • Tested applicationsSuitable for: WB, IP, IHC-P, ICC, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

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

  • Positive control
    • HeLa, MCF 7, SH SY5Y and A549 cell lysates. Human breast carcinoma tissue, Human testis tissue and HeLa cells.
  • General notes

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

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.

Application Abreviews Notes
WB 1/1000 - 1/5000. Predicted molecular weight: 64 kDa.
IP 1/10.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
ICC 1/100 - 1/250.
Flow Cyt 1/50. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Target

  • FunctionGTPase activator for the nuclear Ras-related regulatory protein Ran, converting it to the putatively inactive GDP-bound state.
  • Tissue specificityHighly expressed in brain, thymus and testis.
  • Sequence similaritiesBelongs 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 localizationCytoplasm. 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.
  • Information by UniProt
  • Database links
  • 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

Anti-RanGAP1 antibody [EPR3295] images

  • 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.

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

  • 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.

References for Anti-RanGAP1 antibody [EPR3295] (ab92360)

This product has been referenced in:
  • 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). Read more (PubMed: 23851309) »

See 1 Publication for this product

Product Wall

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.
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Submitted Jul 03 2014

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
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Submitted Jan 23 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
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
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Submitted Jul 01 2013

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"