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Anti-Notch2 antibody (ab8926)

  • Datasheet
  • SDS
Reviews (2)Q&A (8)References (47)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch2 antibody (ab8926)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch2 antibody (ab8926)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch2 antibody (ab8926)

Key features and details

  • Rabbit polyclonal to Notch2
  • Suitable for: IHC-P
  • Reacts with: Human
  • Isotype: IgG

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Overview

  • Product name

    Anti-Notch2 antibody
    See all Notch2 primary antibodies
  • Description

    Rabbit polyclonal to Notch2
  • Host species

    Rabbit
  • Specificity

    The Notch2 protein has many isoforms.  The immunogenic sequence for the anti-Notch2 antibody is identical to the same region in full length (260+ kDa) as well as the cleaved forms ~82 kDa each (Notch 2 extracellular truncation & Notch 2 intracellular domain) and the post translationally modified forms thereof.

  • Tested Applications & Species

    Application Species
    IHC-P
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide corresponding to Human Notch2 aa 1719-1733 (N terminal) (acetyl C) (Cysteine residue). The epitope is only exposed after gamma secretase cleavage and is not accessible in the uncleaved form.
    Sequence:

    CRDASNHKRREPVGQD


    (Peptide available as ab1389, ab205702)
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • General notes

    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 the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.02% Sodium azide
    Constituents: 0.42% Potassium phosphate, 0.88% Sodium chloride
  • Concentration information loading...
  • Purity

    Whole antiserum
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurology process
    • Notch Pathway
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors
    • Stem Cells
    • Signaling Pathways
    • Notch
    • Surface Molecules
    • Stem Cells
    • Signaling Pathways
    • Notch
    • Nuclear
    • Stem Cells
    • Neural Stem Cells
    • Intracellular
    • Stem Cells
    • Neural Stem Cells
    • Surface Molecules

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant Human Notch2 protein (ab114827)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab8926 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
IHC-P
Human
All applications
Mouse
Rat
Application Abreviews Notes
IHC-P
1/500.
Notes
IHC-P
1/500.

Target

  • Function

    Functions as a receptor for membrane-bound ligands Jagged1, Jagged2 and Delta1 to regulate cell-fate determination. Upon ligand activation through the released notch intracellular domain (NICD) it forms a transcriptional activator complex with RBPJ/RBPSUH and activates genes of the enhancer of split locus. Affects the implementation of differentiation, proliferation and apoptotic programs (By similarity). Involved in bone remodeling and homeostasis. In collaboration with RELA/p65 enhances NFATc1 promoter activity and positively regulates RANKL-induced osteoclast differentiation. Positively regulates self-renewal of liver cancer cells (PubMed:25985737).
  • Tissue specificity

    Expressed in the brain, heart, kidney, lung, skeletal muscle and liver. Ubiquitously expressed in the embryo.
  • Involvement in disease

    Alagille syndrome 2
    Hajdu-Cheney syndrome
  • Sequence similarities

    Belongs to the NOTCH family.
    Contains 6 ANK repeats.
    Contains 35 EGF-like domains.
    Contains 3 LNR (Lin/Notch) repeats.
  • Post-translational
    modifications

    Synthesized in the endoplasmic reticulum as an inactive form which is proteolytically cleaved by a furin-like convertase in the trans-Golgi network before it reaches the plasma membrane to yield an active, ligand-accessible form. Cleavage results in a C-terminal fragment N(TM) and a N-terminal fragment N(EC). Following ligand binding, it is cleaved by TNF-alpha converting enzyme (TACE) to yield a membrane-associated intermediate fragment called notch extracellular truncation (NEXT). This fragment is then cleaved by presenilin dependent gamma-secretase to release a notch-derived peptide containing the intracellular domain (NICD) from the membrane.
    Hydroxylated by HIF1AN.
    Can be either O-glucosylated or O-xylosylated at Ser-613 by POGLUT1.
  • Cellular localization

    Cell membrane and Nucleus. Cytoplasm. Following proteolytical processing NICD is translocated to the nucleus. Retained at the cytoplasm by C8orf4 (PubMed:25985737).
  • Target information above from: UniProt accession Q04721 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 4853 Human
    • Entrez Gene: 18129 Mouse
    • Omim: 600275 Human
    • SwissProt: Q04721 Human
    • SwissProt: O35516 Mouse
    • SwissProt: Q9QW30 Rat
    • Unigene: 487360 Human
    • Unigene: 485843 Mouse
    • Alternative names

      • AGS2 antibody
      • hN2 antibody
      • Motch B antibody
      • N2 antibody
      • N2ECD antibody
      • N2ICD antibody
      • neurogenic locus notch homolog protein 2 antibody
      • NOTC2_HUMAN antibody
      • Notch 2 antibody
      • Notch 2 intracellular domain antibody
      • Notch Drosophila homolog 2 antibody
      • Notch homolog 2 antibody
      • Notch homolog 2 Drosophila antibody
      • Notch2 antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch2 antibody (ab8926)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch2 antibody (ab8926)Image from Murta, Daniel et al., PLoS ONE 8.8 (2013): e72767. Fig 2J. doi: 10.1371/journal.pone.0072767 P. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of CD-1 mice testes tissue sections labeling Notch2 with ab8926 at 1/100 dilution. CD-1 mice testes were fixed in 4% neutral phosphate buffered formalin at room temperature for 24 h and, after subsequent dehydration in ethanol, were embedded in paraffin. Tissue sections were stained by haematoxylin and identification of cell types was done through histology; The antigen retrieval step was performed in citrate buffer (10 mM, pH 6.0), Blocking was performed in PBS with 2% w/v bovine serum albumin for one hour at room temperature. Tissue sections were incubated overnight at 4°C with anti-Notch2 (ab8926). Positive immunostaining in brown color. Notch2 is expressed in germ cells entering meiosis.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch2 antibody (ab8926)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch2 antibody (ab8926)Image from Murta, Daniel et al., PLoS ONE 8.8 (2013): e72767. Fig 2J. doi: 10.1371/journal.pone.0072767 P.

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of CD-1 mice testes tissue sections labeling Notch2 with ab8926 at 1/100 dilution. CD-1 mice testes were fixed in 4% neutral phosphate buffered formalin at room temperature for 24 h and, after subsequent dehydration in ethanol, were embedded in paraffin. Tissue sections were stained by haematoxylin and identification of cell types was done through histology; The antigen retrieval step was performed in citrate buffer (10 mM, pH 6.0), Blocking was performed in PBS with 2% w/v bovine serum albumin for one hour at room temperature. Tissue sections were incubated overnight at 4°C with anti-Notch2 (ab8926). Positive immunostaining in brown color. Notch2 is ubiquitously expressed in germ cells.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch2 antibody (ab8926)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch2 antibody (ab8926)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue sections labeling Notch2 with ab8926 at 1/500 dilution. No pre-treatment of sample was required. The image shows the localization of antibody as the precipitated red signal, with a hematoxylin purple nuclear counter stain.

    Protocols

    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (47)

    Publishing research using ab8926? Please let us know so that we can cite the reference in this datasheet.

    ab8926 has been referenced in 47 publications.

    • Zhao J  et al. MicroRNA-204-5p inhibits the osteogenic differentiation of ankylosing spondylitis fibroblasts by regulating the Notch2 signaling pathway. Mol Med Rep 22:2537-2544 (2020). PubMed: 32705191
    • Jiang H  et al. Long non-coding RNA SNHG3 promotes breast cancer cell proliferation and metastasis by binding to microRNA-154-3p and activating the notch signaling pathway. BMC Cancer 20:838 (2020). PubMed: 32883233
    • Song Y  et al. The ethyl acetate extracts of radix trichosanthis protect retinal vascular endothelial cells from high glucose-induced injury. J Ethnopharmacol 240:111954 (2019). PubMed: 31085225
    • Yu Y  et al. MiR-490-5p inhibits the stemness of hepatocellular carcinoma cells by targeting ECT2. J Cell Biochem 120:967-976 (2019). PubMed: 30206962
    • Yang J  et al. Role of Jagged1/STAT3 signalling in platinum-resistant ovarian cancer. J Cell Mol Med 23:4005-4018 (2019). PubMed: 30993885
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 10 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Notch2 antibody

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (Liver progenitor)
    Permeabilization
    Yes - Triton X-100
    Specification
    Liver progenitor
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 21 2017

    Question

    I would like to know if it is possible to get the pre-immune serum of the same rabbit that the one that has been immunized to give antibodies raised against the targeted protein. The antibodies that I'm refering to are: ab7280 Lot. GR17350 ab8926 Lot. GR21934 ab10554 Lot. GR4787 ab23426 Lot. GR15959 ab27526 Lot. GR40329

    Read More

    Abcam community

    Verified customer

    Asked on Apr 15 2013

    Answer


    Unfortunately it is not possible for us to sell the pre-immune serum of our rabbits.
    I am sorry that on this occasion I could not be of more help.

    Read More

    Abcam Scientific Support

    Answered on Apr 15 2013

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Notch 2 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (postnatal day 1 testes)
    Specification
    postnatal day 1 testes
    Fixative
    Paraformaldehyde
    Permeabilization
    No
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Dr. Qing Wen

    Verified customer

    Submitted Dec 19 2012

    Question

    Dear technical team,

    Our customer would like to know the M.W. of ab8926.
    https://www.abcam.com/Notch-2-antibody-ab8926.html

    I checked 'Scientific Support" and told the customer it's 77kda but,
    he said he saw 110 kDa on the references and also checked "specific references".
    So, let me know exact expected molecular weight of this antibody.


    Best regards,

    Read More

    Abcam community

    Verified customer

    Asked on Jun 22 2012

    Answer

    Thank you for your inquiry.

    The immunogen we used was CRDASNHKRREPVGQD, corresponding to amino acids 1719-1733 of Human activated Notch2.

    The Notch 2 intracellular domain is from amino acids 1697 to 2471 and is 775 amino acids long and shows multiple phosphorylation sites. If the protein is phosphorylated the molecular weight will be considerable higher that the calculated weight according to the unmodified sequence.

    Notch is also processed and cleaved in multiple different ways which will also affect the the molecular weight seen on a WB. A literature search will help you customer interpret their data.

    I hope this information is helpful and wish you and your customer a good weekend.

    Read More

    Abcam Scientific Support

    Answered on Jun 22 2012

    Question

    problem with ab14955 in IHC-P, Mouse samples. Would like to test ab8926 on Mouse tissue (untested species)

    Read More

    Abcam community

    Verified customer

    Asked on Nov 09 2011

    Answer

    Thank you for calling us and for alerting us to the problem you are experiencing with our product. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly. As we discussed on the phone, I am attaching our questionnaire so that we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody. About the Notch 2 antibody ab8926 : To our knowledge, ab8926 has not been tested in mouse. Therefore, I can offer a discount off a future purchase if you buy ab8926 now, test it in Mouse and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free primary antibody. If you are interested in this offer, please follow these steps: 1. Reply to this e-mail to let me know that you would like to proceed and test ab8926 in mouse. I will then send a discount code. This code must be issued before purchasing ab8926 so please wait for my reply before ordering. 2. Purchase ab8926 either by phone, fax, or online (www.abcam.com). 3. Test it in mouse. 4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews. 5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any primary antibody ordered and the discount code is valid for 4 months after issue. We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab8926 turns out to be unsuitable for mouse, you will still receive the discount on your next purchase after your Abreview has been submitted. Please let me know if you have any questions about this offer and I would be happy to help you further. The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount. I look forward to receiving your reply with your questionnaire and image(s) for ab14955.

    Read More

    Abcam Scientific Support

    Answered on Nov 09 2011

    Question

    I submitted Abreview about ab8926 with a WB in TIFF file last week. Please check. You may see how GSI affect Notch2 cleavage status. I am actually confused. Let me know what you think ASAP. Thanks!

    Read More

    Abcam community

    Verified customer

    Asked on Mar 01 2006

    Answer

    When I was a postdoc I studied gamma-secretase inhibitors, APP, abeta, Notch etc. so I feel like I can interpret the result. I would have expected the activated Notch2 protein to be ~ 77 kD more or less. 20 uM of most gamma-secretase inhibitors if applied the correct way should inhibit Notch cleavage. What gamma-secretase inhibitor are you using and how did you apply it? What cells are you using? Have you tried blocking in 5% milk for an hour and then incubating in primary overnight instead of only for 2 hours? I would predict a band at about 77 kD and then as you apply more of the GSI the band should disappear. If the antibody also recognized the uncleaved Notch2 then uncleaved Notch2 should accumulate, but this antibody should only recognize activated Notch2. Strangely the review seems to demonstrate that the 73 kD band accumulates and the 23 kD band disappears. Based on MW and this being an activated Notch2 antibody I would think both bands are non-specific, but it is very strange that these two non-specific bands respond to GSI treatment. What is your interpretation? I look forward to your reply.

    Read More

    Abcam Scientific Support

    Answered on Mar 02 2006

    Question

    Hello, Bill: When I used gradient gel for WB, your ab8926 gave me total 3 binds, 2 looks like dimer around 60kDa, the other smaller one around 23kDa, gamma-secretase inhibitor blocked the 2 bigger binds cleavage. But I am not sure the antigen/immunogen you used was only the most active NICD or may be a larger fragment which including part ot transmembrane. Would you tell me the size of protein you would predict using your ab8926? Thanks!

    Read More

    Abcam community

    Verified customer

    Asked on Feb 22 2006

    Answer

    Thank you for your reply. I was a postdoc in Weiming Xia's lab for four years and spent a considerable amount of time conducting research on gamma-secretase so I should be able to help you. The immunogen we used was CRDASNHKRREPVGQD, corresponding to amino acids 1719-1733 of Human activated Notch2. The Notch 2 intracellular domain is from amino acids 1697 to 2471 and is 775 amino acids long. The transmembrane domain is potentially from amino acids 1678 to 1698. Im would have expected the NICD2 band to be ~ 77 kDa, however, I would not be surprised if it ran at a different molecular weight. IF you are seeing a dimer that is inhibited by a gamma-secretase inhibitor then I believe you are seeing the real NICD. Perhaps you see a dimer because we know that gamma-secretase can cleave at several locations and you can pick this up with this antibody. At this time I am uncertain as to the identity of the 23 kDa band. This antibody has only been tested in ELISA against the peptide immunogen, in which a strong response has been detected. This antibody has not been tested in other applications so I cannot really tell you what size band we would expect to see. Can you please submit your results to fast-track@abcam.com so we can determine if the antibody is working properly? I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

    Read More

    Abcam Scientific Support

    Answered on Feb 23 2006

    Question

    Hi Jennifer, Thank you for attending to this matter. We have had such good experiences with the AbCam reagents that I was surprised the last batch failed. I would like to try the newer batch you have available. Do you need any shipping information? We will test it out as soon as we receive it and I'll let you know the results. Thank you once again,

    Read More

    Abcam community

    Verified customer

    Asked on Jul 28 2004

    Answer

    I'm sending you another vial of ab8926 as a replacement. For your record it is order# 48528 and you should receive it later this week or early next. Please do let me know how this batch works out for you. As I mentioned, ab8926 is a Fast Track antibody and thus has not been fully characterized and so your feedback is extremely valuable. Please let me know if you have any more questions.

    Read More

    Abcam Scientific Support

    Answered on Jul 28 2004

    Question

    Customer previously bought a vial of ab8926 back in the fall and it worked good in IHC-P. Recently bought another vial, lot# 46293 (order# 42237), and is getting very high background.

    Read More

    Abcam community

    Verified customer

    Asked on Jul 16 2004

    Answer

    Thank you very much for your patience. As you know, ab8926 is a Fast Track antibody and thus has not been fully characterized. We have not yet had any feedback regarding how this antibody works in applications other than ELISA, and so your feedback is very valuable. It is difficult to determine if the problems that you have experienced with the second batch were due to that specific lot, the antibody, or experimental conditions. We have a newer batch in stock and I can send you a vial of that batch to try. Please do let me know if you would like me to send that out to you.

    Read More

    Abcam Scientific Support

    Answered on Jul 23 2004

    Question

    I am very interested in testing Notch 2 antibody (ab8926) and also Notch 2 (activated) peptide (human) (ab 1389). But before I would like to have more information. So, of course I would like to know if you saw already any cross-reactivity with other species or you tested other applications than ELISA. If your answer for everything is NO, I would like to ask you for other two things: First: If you could provide us with a very small aliquote to test if the antibody recognize N2 in chicken samples, or Second: if you could let me know if somebody has asked for this antibody in order to make in contact with him and know how it is working. Thank you very much for your time. Best regards,

    Read More

    Abcam community

    Verified customer

    Asked on May 01 2003

    Answer

    All the information we have for our Rabbit polyclonal to Notch 2 (activated) is on the datasheet. It has been tested in ELISA against the immunising peptide but so far has not been further tested for species cross reactivity or suitability for use in other applications. We do not offer small samples for testing purposes. Investigative grade antibodies are offered at a reduced price compared with other antibodies in our range. There is a link on the summary page of the online datasheet which will give you more information about Investigative Grade antibodies and I would be most interested in your feedback. To answer your second question, there are currently three other researchers working with this antibody but for reasons of confidentiality, I am unable to pass on their contact details. As you have specified an interest in Notch 2 in your Abcam account, should any of these researchers decide to write a review about this antibody, an update e-mail will automatically be sent to you. Please let me know if you have any comments about our investigative grade antibodies.

    Read More

    Abcam Scientific Support

    Answered on May 02 2003

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    For licensing inquiries, please contact partnerships@abcam.com

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