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Neuroscience Neurology process Notch Pathway

Anti-NUMB抗体(ab4147)

  • Datasheet
  • SDS
Reviews (8)Q&A (6)References (33)

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Western blot - Anti-NUMB antibody (ab4147)
  • Immunocytochemistry/ Immunofluorescence - Anti-NUMB antibody (ab4147)
  • Immunocytochemistry/ Immunofluorescence - Anti-NUMB antibody (ab4147)
  • Immunocytochemistry/ Immunofluorescence - Anti-NUMB antibody (ab4147)

Key features and details

  • Goat polyclonal to NUMB
  • Suitable for: WB, ICC/IF
  • Reacts with: Mouse, Human
  • Isotype: IgG

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概述

  • 产品名称

    Anti-NUMB抗体
    参阅全部 NUMB 一抗
  • 描述

    山羊多克隆抗体to NUMB
  • 宿主

    Goat
  • 特异性

    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.
  • 经测试应用

    适用于: WB, ICC/IFmore details
  • 种属反应性

    与反应: Mouse, Human
    预测可用于: Chicken, Cow, Dog
  • 免疫原

    Synthetic peptide corresponding to Human NUMB aa 638-651 (C terminal).
    Sequence:

    C-PFSSDLQKTFEIEL


    Database link: P49757
    (Peptide available as ab22990)
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • 阳性对照

    • WB: Mouse brain lysate; ICC: A431 and NIH3T3 cells;
  • 常规说明

    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

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

性能

  • 形式

    Liquid
  • 存放说明

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • 存储溶液

    pH: 7.30
    Preservative: 0.02% Sodium azide
    Constituents: Tris buffered saline, 0.5% BSA
  • Concentration information loading...
  • 纯度

    Immunogen affinity purified
  • 纯化说明

    Purified from goat serum by ammonium sulphate precipitation followed by antigen affinity chromatography using the immunizing peptide.
  • 克隆

    多克隆
  • 同种型

    IgG
  • 研究领域

    • Neuroscience
    • Neurology process
    • Notch Pathway
    • Neuroscience
    • Neurology process
    • Neurogenesis
    • Stem Cells
    • Signaling Pathways
    • Notch
    • Cytoplasmic

相关产品

  • Compatible Secondaries

    • Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (ab150129)
    • Donkey Anti-Goat IgG H&L (HRP) (ab205723)
  • Isotype control

    • Goat IgG, polyclonal - Isotype Control (ab37373)
  • Positive Controls

    • A-431 whole cell lysate (ab7909)
  • Recombinant Protein

    • Recombinant Human NUMB protein (ab131949)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab4147于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
WB (1)
Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 80 kDa (predicted molecular weight: 71 kDa).

1 hour primary incubation at room temperature is recommended for this product.

ICC/IF (3)
Use a concentration of 10 µg/ml.
说明
WB
Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 80 kDa (predicted molecular weight: 71 kDa).

1 hour primary incubation at room temperature is recommended for this product.

ICC/IF
Use a concentration of 10 µg/ml.

靶标

  • 功能

    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.
  • 序列相似性

    Contains 1 PID domain.
  • 翻译后修饰

    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.
  • 细胞定位

    Membrane.
  • Target information above from: UniProt accession P49757 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 数据库链接

    • Entrez Gene: 512187 Cow
    • Entrez Gene: 480381 Dog
    • Entrez Gene: 8650 Human
    • Entrez Gene: 18222 Mouse
    • Omim: 603728 Human
    • SwissProt: P49757 Human
    • SwissProt: Q9QZS3 Mouse
    • Unigene: 654609 Human
    • Unigene: 714879 Human
    • Unigene: 4390 Mouse
    see all
  • 别名

    • 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

图片

  • Western blot - Anti-NUMB antibody (ab4147)
    Western blot - Anti-NUMB antibody (ab4147)
    Anti-NUMB antibody (ab4147) at 1 µg/ml + Mouse brain lysates at 35 µg

    Predicted band size: 71 kDa



    Detected by chemiluminescence.

  • Immunocytochemistry/ Immunofluorescence - Anti-NUMB antibody (ab4147)
    Immunocytochemistry/ Immunofluorescence - Anti-NUMB antibody (ab4147)
    Immunocytochemistry/ Immunofluorescence analysis of A431 cells labeling NUMB with ab4147 at 10ug/mL. Alexa Fluor488 at 2ug/mL was used as the secondary antibody. The nuclear stain is DAPI (blue). Negative control: Unimmunized goat IgG (10ug/mL) followed by Alexa Fluor488 secondary antibody (2ug/mL).
     
    Cells were fixed with paraformaldehyde and permeabilized with 0.15% Triton.
  • Immunocytochemistry/ Immunofluorescence - Anti-NUMB antibody (ab4147)
    Immunocytochemistry/ Immunofluorescence - Anti-NUMB antibody (ab4147)

    Immunocytochemistry/ Immunofluorescence analysis of NIH3T3 cells labeling NUMB with ab4147 at 10ug/mL. Alexa Fluor488 at 2ug/mL was used as the secondary antibody. The nuclear stain is DAPI (blue). Negative control: Unimmunized goat IgG (10ug/mL) followed by Alexa Fluor488 secondary antibody (2ug/mL).

    Cells were fixed with paraformaldehyde and permeabilized with 0.15% Triton.

  • Immunocytochemistry/ Immunofluorescence - Anti-NUMB antibody (ab4147)
    Immunocytochemistry/ Immunofluorescence - Anti-NUMB antibody (ab4147)This image is courtesy of an anonymous Abreview

    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

实验方案

  • Recommended WB protocol for ab4147

Click here to view the general protocols

数据表及文件

  • SDS download

  • Datasheet download

    Download

文献 (33)

发表研究结果有使用 ab4147?请让我们知道,以便我们可以引用本数据表中的参考文章。

ab4147 被引用在 33 文献中.

  • Li Y  et al. CUL4B contributes to cancer stemness by repressing tumor suppressor miR34a in colorectal cancer. Oncogenesis 9:20 (2020). PubMed: 32054830
  • Zhao Y  et al. CD82-TRPM7-Numb signaling mediates age-related cognitive impairment. Geroscience 42:595-611 (2020). PubMed: 32088828
  • Cheng C  et al. Numb negatively regulates the epithelial-to-mesenchymal transition in colorectal cancer through the Wnt signaling pathway. Am J Physiol Gastrointest Liver Physiol 318:G841-G853 (2020). PubMed: 32146835
  • Huang G  et al. TUSC7 suppression of Notch activation through sponging MiR-146 recapitulated the asymmetric cell division in lung adenocarcinoma stem cells. Life Sci 232:116630 (2019). PubMed: 31279783
  • Abdi K  et al. EGFR Signaling Termination via Numb Trafficking in Ependymal Progenitors Controls Postnatal Neurogenic Niche Differentiation. Cell Rep 28:2012-2022.e4 (2019). PubMed: 31433979
View all Publications for this product

客户评价及客户问答

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提交评价 提交问题

1-10 of 14 Abreviews or Q&A

Immunocytochemistry/ Immunofluorescence abreview for Anti-NUMB antibody

Excellent
Abreviews
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

提交于 Mar 19 2013

Question

Anti-NUMB antibody for Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) (Human)
Sample Species: Human
Type: Tissue sections
Specification: Skin equivalent
Fixative: Paraformaldehyde
Antigen retrieval step: Heat mediated
Buffer/Enzyme Used: Dako EnVision Kit 5007
Permeabilization: No
Dilution: 1/20
Incubation time: 1 hour 30 minutes at 25°C
Diluent: 3% BSA in PBS
Secondary Antibody: Dako EnVision Kit 5007
Conjugation: HRP

Read More

Abcam community

Verified customer

Asked on Jul 16 2012

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

Abcam Scientific Support

回复于 Jul 16 2012

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-NUMB antibody

Inconclusive
Abreviews
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

提交于 Jul 13 2012

Immunocytochemistry/ Immunofluorescence abreview for Anti-NUMB antibody

Excellent
Abreviews
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

提交于 Jul 06 2012

Question

Dear Tech support

Could you pleasespeculate regarding ab4147 reactivity against Drosophila derived samples?

Would you like to offer a participation in the abreview promotion for Drosophila with one of the Numb antibodies available.


Thank you for your kind help.

Regards

Read More

Abcam community

Verified customer

Asked on Feb 07 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 Scientific Support

回复于 Feb 07 2012

Immunocytochemistry/ Immunofluorescence abreview for Anti-NUMB antibody

Excellent
Abreviews
Abreviews
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
Read More

Abcam user community

Verified customer

提交于 Dec 09 2009

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-NUMB antibody

Poor
Abreviews
Abreviews
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
Read More

Abcam user community

Verified customer

提交于 Oct 26 2007

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-NUMB antibody

Below Average
Abreviews
Abreviews
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%
Read More

Abcam user community

Verified customer

提交于 Jul 13 2007

Western blot abreview for Anti-NUMB antibody

Average
Abreviews
Abreviews
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
Read More

Abcam user community

Verified customer

提交于 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

Abcam community

Verified customer

Asked on Apr 24 2006

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

Abcam Scientific Support

回复于 Apr 27 2006

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