Overview

  • Product nameAnti-NUMB antibody
    See all NUMB primary antibodies
  • Description
    Rabbit polyclonal to NUMB
  • SpecificityThe antibody may cross react with NUMBL protein (Swissprot ID Q9Y6R0) because the immunogen used to raise this antibody has a 92% homology with NUMBL protein from Human and Mouse. Denaturing of the protein sample is imperative to the success of ab14140 detecting a single specific band in WB (denature at 95°C for 15min). An additional, higher band (around 80kDa) is seen if not denatured.
  • Tested applicationsSuitable for: IHC-P, ICC/IF, IHC-Fr, WB, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Zebrafish
    Predicted to work with: Chicken, Monkey, Non Human Primates
  • Immunogen

    Synthetic peptide derived from residues 600 to the C-terminus of Human NUMB.

    .

  • Positive control
    • This antibody gave a positive signal in Human, Mouse and Rat Brain Tissue Lysates.

Properties

Applications

Our Abpromise guarantee covers the use of ab14140 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
IHC-P Use at an assay dependent concentration.
ICC/IF Use a concentration of 8 µg/ml.
IHC-Fr Use at an assay dependent concentration.
WB Use a concentration of 0.5 - 1 µg/ml. Detects a band of approximately 75 kDa (predicted molecular weight: 70 kDa).Can be blocked with Human NUMB peptide (ab14141).
IP Use at an assay dependent concentration.

Target

  • FunctionPlays 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 similaritiesContains 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 localizationMembrane.
  • 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

Anti-NUMB antibody images

  • Image courtesy of Human Protein Atlas

    ab14140 staining NUMB in Human adrenal gland. The paraffin embedded human tissue was incubated with ab14140 (1/250 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6. Ab14140 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines.

    Further results for this antibody can be found at www.proteinatlas.org

  • NUMB was immunoprecipitated using 0.5mg Mouse Brain whole tissue lysate, 5µg of Rabbit polyclonal to NUMB and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Mouse Brain whole tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab14140.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 75kDa: NUMB.
  • ab14140 detecting NUMB protein in dissociated neural precursor cells cultured from embryonic day 13 mouse neocortex in the presence of bFGF, EGF and LIF. NUMB immunoreactivity (green) was found in the cytosol of neural precursor cells. This localisation is in general agreement with published studies (e.g. Chen et al., EJN, 2005), where similar labelling is observed in the cytosol of cells of the grey matter of adult mouse spinal cord. Immunocytochemistry: All steps were performed in PBS. Cells were fixed in 4% PFA for 15min, permeabilised with 0.1% TX100 for 10min and blocked with 5% BSA, 0.1% TX100 for 45min. ab14140 was incubated at 8µg/ml for 12h at 4°C in 5% BSA, 0.1% TX100. Cultures were washed (3x) of primary antibody solution. Goat anti-rabbit AlexaFluor 488 was used as secondary antibody (1/400) in 5% BSA, 0.1% TX100 for 1h at RT. To-pro-3 was used as a nuclear counterstain (blue). Treated cultures were mounted on glass coverslips with Mowiol.

  • ab14140 at 1/100 staining rat brain tissue sections by IHC-Fr. The tissue was paraformaldehyde fixed and blocked with 5% serum prior to incubation with the antibody for 24 hours. An Alexa-Fluor ® 488 conjugated goat anti-rabbit antibody was used as the secondary. NUMB staining is shown in green.

    See Abreview

  • All lanes : Anti-NUMB antibody (ab14140) at 1 µg/ml

    Lane 1 : Brain (Rat) Tissue Lysate
    Lane 2 : Brain (Mouse) Tissue Lysate
    Lane 3 : Human brain tissue lysate - total protein (ab29466)

    Lysates/proteins at 10 µg per lane.

    Secondary
    Anti-Rabbit IgG VHH Single Domain Antibody (HRP) (ab191866) at 1/50000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 70 kDa
    Observed band size : 75 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 74 kDa (possible isoform),77 kDa (possible post-translational modification).

    Exposure time : 4 minutes

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab14140 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

  • Anti-NUMB antibody (ab14140) at 0.5 µg/ml + Adult Rat whole brain lysate at 40 µg

    Secondary
    Goat anti-rabbit IgG HRP at 1/20000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 70 kDa
    Observed band size : 75 kDa (why is the actual band size different from the predicted?)


    Exposure time : 30 seconds

    This image is courtesy of Randal Moldrich, CNRS UMR7637, ESPCI, France

     Denaturing of the protein sample was imperative to the success of this WB (denatured at 95°C for 15min). An additional, higher band (around 80kDa) is seen if not denatured. Original concentration tried was 1ug/ml but we found this high given the circumstances and have since used 0.5ug/ml. Theoretically, even less could be used.

  • ab14140 staining NUMB in Zebrafish embryo nervous system by Immunohistochemistry (Whole mount). Samples were incubated with primary antibody (1/500 in BSA blocking buffer) for 48 hours at 4°C. An Alexa Fluor®488-conjugated Goat anti-rabbit  polyclonal (1/500) was used as the secondary antibody.

    See Abreview

  • Anti-NUMB antibody (ab14140) at 0.5 µg/ml + Mouse whole brain lysate

    Secondary
    Goat anti-rabbit IgG HRP at 1/20000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 70 kDa
    Observed band size : 75 kDa (why is the actual band size different from the predicted?)


    Exposure time : 30 seconds

    This image is courtesy of Randal Moldrich, CNRS UMR7637, ESPCI, France

    Denaturing of the protein sample was imperative to the success of this WB. An additional, higher band (around 80kDa) is seen if not denatured. Original concentration tried was 1ug/ml but we found this high given the circumstances and have since used 0.5ug/ml. Theoretically, even less could be used.


  • Performed under reducing conditions.

    Predicted band size : 70 kDa
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  • Performed under reducing conditions.

    Predicted band size : 70 kDa
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References for Anti-NUMB antibody (ab14140)

This product has been referenced in:
  • Kudinov AE  et al. Musashi-2 (MSI2) supports TGF-ß signaling and inhibits claudins to promote non-small cell lung cancer (NSCLC) metastasis. Proc Natl Acad Sci U S A 113:6955-60 (2016). WB . Read more (PubMed: 27274057) »
  • Zhao C  et al. Numb family proteins are essential for cardiac morphogenesis and progenitor differentiation. Development 141:281-95 (2014). Read more (PubMed: 24335256) »

See all 14 Publications for this product

Product Wall

Application IHC - Wholemount
Sample Zebrafish Embryo (Nervous system)
Specification Nervous system
Username

Abcam user community

Verified customer

Submitted Nov 03 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Pancreas)
Specification Pancreas
Fixative Methanol
Permeabilization No
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted May 07 2013

Thank you for your email.

We had sent you the following information based on your inquiry about the homology between NUMB and NUMBL:

The overall homology between human NUMB and NUMBL proteins is only 42-67%.

But for the ...

Read More

Thank you for contacting us.

The overall homology between human NUMB and NUMBL proteins is only 42-67%.

But for the immunogen sequence, the homology is 93%, unfortunately. Thus, crossreactivity is likely. I'll have thelab look into...

Read More

Thank you for contacting us.

Could you please let me know the NUMBL sequence or the according database entry so that I can check sequence homology for between NUMB and NUMBL in the immunogen region?

Thank you for your help!
<...

Read More
Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Chicken Tissue sections (Spinal cord)
Specification Spinal cord
Fixative Paraformaldehyde
Antigen retrieval step None
Permeabilization Yes - 0.1% Triton-X-100
Blocking step Serum as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 20°C
Username

David Rousso

Verified customer

Submitted Sep 09 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (embryo brain)
Specification embryo brain
Fixative Paraformaldehyde
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 6%
Username

Abcam user community

Verified customer

Submitted May 22 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Rat Tissue sections (Brain)
Specification Brain
Fixative Paraformaldehyde
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Username

Mr. Darius Gleason

Verified customer

Submitted Feb 01 2007

Thank you for your patience. Again, I'm sorry to hear you are having a problem with ab14955. I will do my best to troubleshoot with you. In my last e-mail, I suggested the following modifications to your protocol: 1) Reduce primary antibody - ...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (Whole brain lysate - adult)
Loading amount 40 µg
Specification Whole brain lysate - adult
Treatment Denatured at 95C for 15min
Blocking step Milk as blocking agent for 20 minute(s) · Concentration: 5%
Username

Dr. Randal Moldrich

Verified customer

Submitted May 18 2006

1-10 of 12 Abreviews or Q&A

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