Overview

  • Product name
  • Description
    Goat polyclonal to NUMB
  • Host species
    Goat
  • Specificity
    This antibody is expected to recognize all reported isoforms (NP_001005743.1, 001005744.1, NP_003735, NP_001005745.1). It is also likely to cross-react with Numblike (NUMBL, GeneID: 9253) which has a very similar C-Terminus, although this has not yet been tested experimentally.
  • Tested applications
    Suitable for: IHC-P, WB, IP, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Human
    Predicted to work with: Cow, Dog
  • Immunogen

    Synthetic peptide: PFSSDLQKTFEIEL, corresponding to C terminal amino acids 638-651 of Human NUMB.

  • Positive control
    • A431 lysate. Mouse brain lysates.
  • General notes
    This product may cross-react with Numblike (NUMBL, GeneID: 9253) Principal Names - NUMB; S171; numb homolog (Drosophila) Official Gene Symbol - NUMB GenBank Accession Number – NP_003735 LocusLink ID - 8650 (human); 18222 (mouse) Gene Ontology terms - integral to plasma membrane

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer
    pH: 7.30
    Preservative: 0.02% Sodium azide
    Constituents: 0.5% BSA, Tris buffered saline
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Purification notes
    Purified from goat serum by ammonium sulphate precipitation followed by antigen affinity chromatography using the immunizing peptide.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab4147 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. See Abreview
WB Use a concentration of 0.03 - 0.1 µg/ml. Detects a band of approximately 80 kDa (predicted molecular weight: 71 kDa).Can be blocked with Human NUMB peptide (ab22990).
IP 1/1000. For 1 hr on mouse brain lysate (see Abreview).
ICC/IF Use at an assay dependent dilution. PubMed: 17127749

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

  • ab4147 staining NUMB in the NIH3T3 cell line from Mouse fibroblasts by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and blocked with 10% serum for 60 minutes at 24°C. Samples were incubated with primary antibody (1/100) for 16 hours at 4°C. An  Alexa Fluor®488-conjugated Donkey anti-rabbit polyclonal(1/500) was used as the secondary antibody.

    See Abreview

  • Western Blot analysis of rat brain lysate (35µg protein in RIPA buffer)labeling NUMB with ab4147 at 0.01µg/ml. Primary incubation was 1 hour. Detected by chemiluminescence.

  • Single collagenase-isolated mouse muscle fibre stained with DAPI (blue) to reveal nuclei and anti-Numb (green) which detects satellite cells.

    This picture was kindly supplied as part of the review submitted by Christopher Mann.

  • ab4147 staining (0.03µg/ml) of Mouse Brain lysate (RIPA buffer, 30µg total protein per lane). Primary incubated for 1 hour. Detected by western blot using chemiluminescence.

References

This product has been referenced in:
  • García-Heredia JM & Carnero A NUMB and NUMBL differences in gene regulation. Oncotarget 9:9219-9234 (2018). Read more (PubMed: 29507685) »
  • Tremblay CS  et al. Restricted cell cycle is essential for clonal evolution and therapeutic resistance of pre-leukemic stem cells. Nat Commun 9:3535 (2018). Read more (PubMed: 30166543) »
See all 22 Publications for this product

Customer reviews and Q&As

1-10 of 14 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (NIH3T3)
Specification
NIH3T3
Fixative
Paraformaldehyde
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Mar 19 2013

Answer

Thank you for submitting an Abreview of ab4147. As you may have noticed, your review has now been published on our website.

Since you obtained poor results using the antibody in a tested species and application, we would like to follow up on this to see if we can possibly improve the results you are seeing with this antibody.

Based on the information in your review, I feel that altering the following step in your protocol may help improve your results:

- testing various titers of primary antibody
- a different antigen retrieval solution


If you decide to follow these suggestions, please let me know if they are helpful. Additionally, this antibody is covered by our Abpromise, so if the suggestions do not help and the antibody was purchased within the past 6 months, I would be happy to discuss a refund or replacement with you to resolve this issue.

I look forward to your reply.

Read More
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Skin equivalent)
Specification
Skin equivalent
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Dako EnVision Kit 5007
Permeabilization
No
Blocking step
(agent) for 20 minute(s) · Concentration: 100% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jul 13 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (skin equivalent, human keratinocytes Wholemount)
Specification
skin equivalent, human keratinocytes Wholemount
Fixative
Formaldehyde
Permeabilization
Yes - 0.2% Triton-X100
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jul 06 2012

Answer

Thank you for contacting us.


The anti-NUMB antibody (ab4147) is raised against a peptide corresponding to residues 638-651 of human NUMB. Through sequence alignment of the Drosophila melanogaster protein (SwissProt reference http://www.uniprot.org/uniprot/P16554), this region shares very little homology with the human protein. I would therefore not suggest usign this antibody to detect the Drosophila protein.


Unfortunately we have no other anti-NUMB antibodies that I can suggest as an alternative. You may have more luck by searching Biocompare:


http://www.biocomplare.com


I am sorry that I have not been able to be of more help in this instance. If you have any further questions please do not hesitate to contact us again.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (embryonic brain tissue)
Specification
embryonic brain tissue
Fixative
Paraformaldehyde
Permeabilization
Yes - 1% TX100
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Dec 09 2009

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (embryo CNS)
Specification
embryo CNS
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10 mM sodiumcitrate pH 6.0, also Tris-EDTA pH 9.0 tried
Permeabilization
Yes - 0.1 % Tween-20 for 10-30 minutes
Blocking step
!% BSA and 10%donkey serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Oct 26 2007

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (embryo)
Specification
embryo
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 6%

Abcam user community

Verified customer

Submitted Jul 13 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (embryo brain)
Loading amount
30 µg
Specification
embryo brain
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jun 05 2007

Question

BATCH NUMBER 141307 ORDER NUMBER 96008 DESCRIPTION OF THE PROBLEM Wrong band size: band size is supposed to be approx 71kDa, but I am getting a very robust band at about 115kDa that is much stronger than the band I see at about 70kDa when looking at various mouse cell lysates (see attached file, ab4147 gel on left). When I probe same blot with another Numb antibody (see gel on right), I get much more specificity towards the 70kDa bands. Though I still see a faint band at a larger molecular weight, this is greatly reduced and rather negligible when compared to the robustness of the 70kDa band. Contrarily, the abcam antobody shows the STRONGEST sensitivity to the 115kDa band. Thus, it appears as if the abcam is picking up a non-specific protein in mouse cells (as I mentioned, an abcam technician told me that abcam had only tested this antibody for Westerns on human cell lines), and that this abnormal band is not a product of an abnormal gel or a high level of modification to Numb, as an alternative Numb antibody does not pick up this product. Note: two bands seen at approx 70kDa size are due to four isoforms (65, 66, 71 and 72kDa expacted band sizes). SAMPLE For both gels, lanes are as follows: 1 - E15 forebrain negative control 2 - 3T3 cell line (mouse fibroblasts) negative control 3 - MHP36 cell line (mouse hippocampal stem cell line) negative control 4 - E15 positive 5 - 3T3 positive 6 - MHP36 positive PRIMARY ANTIBODY ab4147 used at 1:200 in 5% nonfat milk/TBST, 2 hrs shaking at RT, followed by 4 x 5 min wash in TBST.. DETECTION METHOD ECL - Lumilight Kit POSITIVE AND NEGATIVE CONTROLS USED E15 - as positive control Lanes 1-3 as negative controls (no primary antibody added) ANTIBODY STORAGE CONDITIONS Aliquotted and stored at -20oC SAMPLE PREPARATION Cells lysed in lysis buffer (Tris pH7.5 50mM, NaCl 150mM, Triton X-100 1%, EDTA 5mM) for 30 min on ice, spun and then supernatant isolated. Mixed with sample buffer in 3:4 ratio, incubated ar 90oC for 6 min and loaded into gel. AMOUNT OF PROTEIN LOADED E15 - 5ul 3T3 - 20ul MHP36 - 10ul ELECTROPHORESIS/GEL CONDITIONS 8% SDS gel TRANSFER AND BLOCKING CONDITIONS Gel transfered for 2.5 hrs at 10V. Block 1 hr in 5% nonfat milk/TBST shaking at RT. SECONDARY ANTIBODY Secondary - Donkey anti-goat IgG HRP, [another company], used 1:2000, 1hr at RT shaking, followed by 4 x 5 min wash in TBST. HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? Have also tried on 10% SDS gel and with incubating primary overnight at 4oC with same results. ADDITIONAL NOTES Also ordered same antibody on purchase order number 90419 (have ordered two vials total). Have spoken with Hugh Spotswood with regards to this matter. As this antibody was advertised as being specific for mouse and does not appear to be, I would greatly appreciate a reimbursement for the two purchases of this antibody. Thank you.

Read More
Answer

Thank you for your enquiry and for providing me with details of the western blotting experiments that you have performed. From the internal notes that I have I can see that mouse reactivity by this antibody was added as a result of two independent Abreviews. Customer feedback following reviews of how the antibody behaves by IP and IHC on formalin-fixed paraffin-embedded sections resulted in mouse being added to the antibody's datasheet. I have updated our datasheet to include the information that you have provided me with. I am prepared to offer you credit against the purchase of this antibody given that it has not behaved as detailed on our datasheet. This is provided that the purchase was made within the past 90 days. If this is the case please e-mail me details of the order and date of purchase and I will ask our accounts team to raise credit against your initial purchase.

Read More

1-10 of 14 Abreviews or Q&A

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