Overview

  • Product name

    Anti-RBPJK antibody [EPR13479]
    See all RBPJK primary antibodies
  • Description

    Rabbit monoclonal [EPR13479] to RBPJK
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human RBPJK aa 100-200. The exact sequence is proprietary.
    Database link: Q06330

  • Positive control

    • WB: 293 cells, MCF7, Raji, F9, C9, and HeLa cell line lysates; ICC/IF: 293T cells; FC: HEK-293;
  • General notes

     

     

    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 ab180588 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 - 1/10000. Detects a band of approximately 60 kDa (predicted molecular weight: 56 kDa).
ICC/IF 1/100.
Flow Cyt 1/100.

Target

  • Function

    Transcriptional regulator that plays a central role in Notch signaling, a signaling pathway involved in cell-cell communication that regulates a broad spectrum of cell-fate determinations. Acts as a transcriptional repressor when it is not associated with Notch proteins. When associated with some NICD product of Notch proteins (Notch intracellular domain), it acts as a transcriptional activator that activates transcription of Notch target genes. Probably represses or activates transcription via the recruitment of chromatin remodeling complexes containing histone deacetylase or histone acetylase proteins, respectively. Specifically binds to the immunoglobulin kappa-type J segment recombination signal sequence.
  • Sequence similarities

    Belongs to the Su(H) family.
    Contains 1 IPT/TIG domain.
  • Cellular localization

    Nucleus. Cytoplasm. Mainly nuclear, upon interaction with RITA/C12orf52, translocates to the cytoplasm, down-regulating the Notch signaling pathway.
  • Information by UniProt
  • Database links

  • Alternative names

    • AI843960 antibody
    • AOS3 antibody
    • CBF 1 antibody
    • CBF-1 antibody
    • CBF1 antibody
    • csl antibody
    • IGKJRB antibody
    • IGKJRB1 antibody
    • J kappa recombination signal binding protein antibody
    • J kappa-recombination signal-binding protein antibody
    • KBF2 antibody
    • NY REN 30 antigen antibody
    • RBP J antibody
    • RBP J kappa antibody
    • RBP JK antibody
    • RBP-J antibody
    • RBP-J kappa antibody
    • RBP-JK antibody
    • Rbpj antibody
    • RBPJK antibody
    • RBPSUH antibody
    • recombination signal binding protein for immunoglobulin kappa J region antibody
    • Recombining binding protein suppressor of hairless antibody
    • Renal carcinoma antigen NY-REN-30 antibody
    • SUH antibody
    • SUH_HUMAN antibody
    see all

Images

  • All lanes : Anti-RBPJK antibody [EPR13479] (ab180588) at 1/1000 dilution (Purified)

    Lane 1 : Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysates at 15 µg
    Lane 2 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates at 15 µg
    Lane 3 : F9 (Mouse embryonal carcinoma epithelial cell) whole cell lysates at 15 µg
    Lane 4 : C6 (Rat glial tumor glial cell) whole cell lysates at 20 µg

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

    Predicted band size: 56 kDa
    Observed band size: 56-60 kDa
    why is the actual band size different from the predicted?



    The double bands are consistent with what has been described in the literature PMID: 9102301 and 24603501

  • Immunocytochemistry/ Immunofluorescence analysis of 293T (Human embryonic kidney epithelial cell) cells labeling RBPJK with Purified ab180588 at 1:100 dilution (10 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) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
  • Flow Cytometry analysis of HEK-293 (Human embryonic kidney epithelial cell) cells labeling RBPJK with Purified ab180588 at 1:100 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. 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).
  • All lanes : Anti-RBPJK antibody [EPR13479] (ab180588) at 0.4 µg/ml (Unpurified)

    Lane 1 : C6 (Rat glial tumor glial cell) whole cell lysate
    Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
    Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
    Lane 4 : Raw264.7 (Mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 56 kDa



    Blocking and diluting buffer: 5% NFDM/TBST. 

    Exposure time: Lane 1-3: 3 minutes
                             Lane 4: 45 seconds

  • Immunofluorescent analysis of 4% paraformaldehyde fixed 293 cells labeling RBPJK with unpurified ab180588 at 1/100 (green). Dapi shown in blue.

  • All lanes : Anti-RBPJK antibody [EPR13479] (ab180588) at 1/1000 dilution (Unpurified)

    Lane 1 : MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysate 20ug prepared using RIPA lysis method
    Lane 2 : MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysate 20ug prepared using 1% SDS hot lysis method

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/20000 dilution

    Predicted band size: 56 kDa
    Observed band size: 60 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Diluting buffer and concentration 5% NFDM/TBST.

    Extra band at: 90kDa
    We are unsure as to the identity of this extra band.

    For Lysate preparation protocol, please refer to the protocol book in the protocol section and/or here (downloadable copy).

  • All lanes : Anti-RBPJK antibody [EPR13479] (ab180588) at 1/2000 dilution (Unpurified)

    Lane 1 : 293 cell line lysate
    Lane 2 : MCF7 cell line lysate
    Lane 3 : HeLa cell line lysate

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 56 kDa
    Observed band size: 60 kDa why is the actual band size different from the predicted?

References

This product has been referenced in:

  • Liang Y  et al. Low expression of NCOA5 predicts poor prognosis in human cervical cancer and promotes proliferation, migration, and invasion of cervical cancer cell lines by regulating notch3 signaling pathway. J Cell Biochem 120:6237-6249 (2019). Read more (PubMed: 30335900) »
  • Zeng WX  et al. Hypertonic saline attenuates expression of Notch signaling and proinflammatory mediators in activated microglia in experimentally induced cerebral ischemia and hypoxic BV-2 microglia. BMC Neurosci 18:32 (2017). Read more (PubMed: 28288585) »
See all 3 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Permeabilization
Yes - Triton X
Specification
HeLa
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jan 19 2017

Application
Western blot
Sample
Human Cell lysate - other (HEK 293 cells)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
30 µg
Specification
HEK 293 cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Jan 18 2017

Application
Western blot
Loading amount
40 µg
Gel Running Conditions
Reduced Denaturing (4%-12%)
Sample
Human Cell lysate - whole cell (Human Umbilical Artery Smooth Muscle Cells)
Specification
Human Umbilical Artery Smooth Muscle Cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Feb 24 2014

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