Overview

  • Product name

    Anti-NUMB antibody [EPR21988]
    See all NUMB primary antibodies
  • Description

    Rabbit monoclonal [EPR21988] to NUMB
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, IP, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human NUMB aa 400 to the C-terminus. The exact sequence is proprietary.
    Database link: P49757

  • Positive control

    • WB: HeLa, Ramos, SK-OV-3, PC-12 and NIH/3T3 cell lysate; Mouse brain and lung lysate; Rat brain lysate. IHC-P: Human breast and kidney tissue; Mouse and rat kidney tissue. Flow cytometry: Ramos cells. IP: HeLa whole cell lysate.
  • 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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab220362 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. Predicted molecular weight: 71 kDa.
IHC-P 1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IP 1/30.
Flow Cyt 1/600.

Target

  • Function

    Plays a role in the process of neurogenesis. Required throughout embryonic neurogenesis to maintain neural progenitor cells, also called radial glial cells (RGCs), by allowing their daughter cells to choose progenitor over neuronal cell fate. Not required for the proliferation of neural progenitor cells before the onset of neurogenesis. Also involved postnatally in the subventricular zone (SVZ) neurogenesis by regulating SVZ neuroblasts survival and ependymal wall integrity. May also mediate local repair of brain ventricular wall damage.
  • Sequence similarities

    Contains 1 PID domain.
  • Post-translational
    modifications

    Isoform 1 and isoform 2 are ubiquitinated by LNX leading to their subsequent proteasomal degradation (By similarity). Ubiquitinated; mediated by SIAH1 and leading to its subsequent proteasomal degradation.
  • Cellular localization

    Membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • c14_5527 antibody
    • C14orf41 antibody
    • FLJ31314 antibody
    • h Numb antibody
    • h-Numb antibody
    • Numb antibody
    • Numb homolog (Drosophila) antibody
    • Numb homolog antibody
    • Numb protein homolog antibody
    • NUMB_HUMAN antibody
    • Protein numb homolog antibody
    • Protein S171 antibody
    • S171 antibody
    see all

Images

  • All lanes : Anti-NUMB antibody [EPR21988] (ab220362) at 1/1000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Rat brain lysate
    Lane 3 : Mouse lung lysate
    Lane 4 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate
    Lane 5 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 71 kDa



    Blocking/diluting buffer and concentration: 5% NFDM/TBST 

    Exposure time: Line 1-2: 3 minutes; Line 3: 93 seconds; Line 4-5: 3 minutes.

     

  • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized Ramos (Human Burkitt's lymphoma B lymphocyte) cell line labeling NUMB with ab220362 at 1/600 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells incubated with secondary antibody only) (blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

  • NUMB was immunoprecipitated from 0.35 mg HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate with ab220362 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab220362 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/5000 dilution.
    Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10 μg(Input).
    Lane 2: ab220362 IP in HeLa whole cell lysate (+).
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab220362 in HeLa whole cell lysate (-).


    Blocking/Dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 3 minutes.

  • Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling NUMB with ab220362 at 1/1000 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rar kidney tubules (PMID:27016419) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). 

    Perform heat-mediated antigen retrieval using Universal HIER antigen retrieval reagents (10X)(ab208572).

  • Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling NUMB with ab220362 at 1/1000 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse kidney tubules (PMID:27016419) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). 

    Perform heat-mediated antigen retrieval using Universal HIER antigen retrieval reagents (10X)(ab208572).

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling NUMB with ab220362 at 1/1000 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human kidney tubules (PMID:27016419) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). 

    Perform heat-mediated antigen retrieval using Universal HIER antigen retrieval reagents (10X)(ab208572).

  • Immunohistochemical analysis of paraffin-embedded human breast tissue labeling NUMB with ab220362 at 1/1000 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human breast tissue (PMID: 15492044) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). 

    Perform heat-mediated antigen retrieval using Universal HIER antigen retrieval reagents (10X)(ab208572).

  • All lanes : Anti-NUMB antibody [EPR21988] (ab220362) at 1/1000 dilution

    Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
    Lane 2 : SK-BR-3 (human breast adenocarcinoma epithelial cell) whole cell lysate
    Lane 3 : T-47D (human ductal breast epithelial tumor epithelial cell) whole cell lysate
    Lane 4 : Ramos (human Burkitt's lymphoma B lymphocyte) whole cell lysate
    Lane 5 : SK-OV-3 (human ovarian cancer epithelial cell) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 71 kDa



    Blocking/diluting buffer and concentration: 5% NFDM/TBST.

    Exposure time: Line 1-3: 3 minutes; Line 4: 15 seconds; Line 5: 37 seconds.

    Negative controls: SK-BR-3, T-47D (PMID: 20443831).

References

ab220362 has not yet been referenced specifically in any publications.

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