Recombinant
RabMAb

Recombinant Anti-c-Myb (phospho S11) antibody [EP769Y] - ChIP Grade (ab45150)

Rabbit recombinant monoclonal c-Myb (phospho S11) antibody [EP769Y]. Validated in WB, IP, IHC, ChIP, Dot, ICC/IF and tested in Human. Cited in 28 publication(s). Independently reviewed in 1 review(s).

Overview

  • Product name

    Anti-c-Myb (phospho S11) antibody [EP769Y] - ChIP Grade
    See all c-Myb primary antibodies
  • Description

    Rabbit monoclonal [EP769Y] to c-Myb (phospho S11) - ChIP Grade
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, IP, Dot blot, ChIPmore details
    Unsuitable for: Flow Cyt
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human c-Myb aa 1-100 (N terminal). The exact sequence is proprietary.
    Database link: P10242

  • Positive control

    • WB: Ramos, Molt-4, and HL-60 cell lysate. HeLa cell lysate untreated and treated with lambda phosphatase. IHC-P: Human cervical carcinoma tissue.
  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    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 ab45150 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 - 1/5000. Predicted molecular weight: 72 kDa.
IHC-P Use at an assay dependent concentration.
ICC/IF 1/100 - 1/250.
IP 1/50.
Dot blot 1/1000.
ChIP Use at an assay dependent concentration. PubMed: 21296997
  • Application notes
    Is unsuitable for Flow Cyt.
  • Target

    • Function

      Transcriptional activator; DNA-binding protein that specifically recognize the sequence 5'-YAAC[GT]G-3'. Plays an important role in the control of proliferation and differentiation of hematopoietic progenitor cells.
    • Sequence similarities

      Contains 3 HTH myb-type DNA-binding domains.
    • Domain

      Comprised of 3 domains; an N-terminal DNA-binding domain, a centrally located transcriptional activation domain and a C-terminal domain involved in transcriptional repression.
    • Post-translational
      modifications

      Ubiquitinated; mediated by SIAH1 and leading to its subsequent proteasomal degradation.
      Phosphorylated by NLK on multiple sites, which induces proteasomal degradation.
    • Cellular localization

      Nucleus.
    • Information by UniProt
    • Database links

    • Alternative names

      • C myb antibody
      • c myb protein (140 AA) antibody
      • c-myb protein (140 AA) antibody
      • c-myb_CDS antibody
      • c-myb10A_CDS antibody
      • c-myb13A_CDS antibody
      • c-myb14A_CDS antibody
      • c-myb8B_CDS antibody
      • Cmyb antibody
      • efg antibody
      • Myb antibody
      • MYB proto-oncogene transcription factor antibody
      • MYB_HUMAN antibody
      • ONCOGENE AMV antibody
      • Oncogene MYB antibody
      • Proto oncogene c Myb antibody
      • Proto-oncogene c-Myb antibody
      • Transcriptional activator Myb antibody
      • v myb avian myeloblastosis viral oncogene homolog antibody
      • v-myb avian myeloblastosis viral oncogene homolog antibody
      see all

    Images

    • All lanes : Anti-c-Myb (phospho S11) antibody [EP769Y] - ChIP Grade (ab45150) at 1/2000 dilution (purified)

      Lane 1 : Molt-4 (Human lymphoblastic leukemia cell line) cell lysate with c-Myb (phospho S11) peptide
      Lane 2 : Molt-4 cell lysate with c-Myb (phospho S12) peptide
      Lane 3 : Molt-4 cell lysate with c-Myb (phospho S11/12) peptide
      Lane 4 : Molt-4 cell lysate with c-Myb unmodified peptide

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP goat anti-rabbit IgG (H+L) at 1/2000 dilution

      Predicted band size: 72 kDa
      Observed band size: 77 kDa
      why is the actual band size different from the predicted?


      Exposure time: 30 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Immunohistochemical analysis of paraffin-embedded human vervical carcinoma tissue staining c-Myb with unpurified ab45150.

       

    • Dot blot analysis of c-Myb (phospho S11/12) peptide (Lane 1) and c-Myb unmodified peptide (Lane 2) labeling c-Myb (phospho S11) with purified ab45150 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at a dilution of 1/2500.

      Blocking and dilution buffer: 5% NFDM/TBST.

      Exposure time: 3 minutes.

    • All lanes : Anti-c-Myb (phospho S11) antibody [EP769Y] - ChIP Grade (ab45150) at 1/2000 dilution (purified)

      Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate with c-Myc (phospho S11) peptide
      Lane 2 : HeLa cell lysate with c-Myc (phospho S12) peptide
      Lane 3 : HeLa cell lysate with c-Myc (phospho S11/12) peptide
      Lane 4 : HeLa cell lysate with c-Myc unmodified peptide

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (HRP goat anti-rabbit IgG (H+L))

      Predicted band size: 72 kDa
      Observed band size: 75,77 kDa why is the actual band size different from the predicted?



      Blocking/Dilution buffer: 5% NFDM/TBST.

    • All lanes : Anti-c-Myb (phospho S11) antibody [EP769Y] - ChIP Grade (ab45150) at 1/5000 dilution (purified)

      Lane 1 : Untreated Ramos (Human Burkitt's lymphoma cell line) cell lysate
      Lane 2 : Ramos cell lysate, membrane treated with alkaline phosphatase

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP goat anti-rabbit IgG (H+L) at 1/2000 dilution

      Predicted band size: 72 kDa
      Observed band size: 75,77 kDa why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • All lanes : Anti-c-Myb (phospho S11) antibody [EP769Y] - ChIP Grade (ab45150) at 1/5000 dilution (purified)

      Lane 1 : Untreated HL-60 (Human promyelocytic leukemia cell line) cell lysate
      Lane 2 : HL-60 cell lysate, membrane treated with alkaline phosphatase

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP goat anti-rabbit IgG (H+L) at 1/2000 dilution

      Predicted band size: 72 kDa
      Observed band size: 77 kDa why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • All lanes : Anti-c-Myb (phospho S11) antibody [EP769Y] - ChIP Grade (ab45150) at 1/2000 dilution (purified)

      Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
      Lane 2 : HeLa cell lysate, the membrane treated with lambda phosphatase

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/5000 dilution (HRP goat anti-rabbit IgG (H+L))

      Predicted band size: 72 kDa
      Observed band size: 75,77 kDa why is the actual band size different from the predicted?


      Exposure time: 30 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Anti-c-Myb (phospho S11) antibody [EP769Y] - ChIP Grade (ab45150) at 1/10000 dilution (unpurified) + Ramos (Human Burkitt's lymphoma cell line) membrane lysate

      Predicted band size: 72 kDa
      Observed band size: 72,75 kDa why is the actual band size different from the predicted?

    References

    This product has been referenced in:

    • Xu LH  et al. MYB promotes the growth and metastasis of salivary adenoid cystic carcinoma. Int J Oncol 54:1579-1590 (2019). Read more (PubMed: 30896785) »
    • Spagnuolo M  et al. Transcriptional activation of the miR-17-92 cluster is involved in the growth-promoting effects of MYB in human Ph-positive leukemia cells. Haematologica 104:82-92 (2019). Read more (PubMed: 30076175) »
    See all 40 Publications for this product

    Customer reviews and Q&As

    1-3 of 3 Abreviews or Q&A

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (chronic myelogenous leukemia & monkey kidney cells)
    Gel Running Conditions
    Reduced Denaturing (4-12% Bis-Tris (BioRad))
    Loading amount
    50000 cells
    Specification
    chronic myelogenous leukemia & monkey kidney cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

    Dr. Ragnhild Eskeland

    Verified customer

    Submitted Apr 22 2015

    Answer

    Thank you for contacting us.

    This product is raised against the N-terminal region. It should not be affected by a truncated C-terminus. We do guarantee this to recognizeAvian myeloblastosis viral (v Myb) in human in the following applications; WB,IHC-P,ICC/IF,IP,ChIP.

    For UNTESTED applications, we have established a testing discount program. Here is a brief description of how it works:

    The testing discount program is for customers who like to use an antibody/protein on an untested species/application. You would purchase the antibody at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount equal to the value of the product tested. The terms and conditions of this offer can be found here: https://www.abcam.com/AbTrial


    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Answer

    Vielen Dank für Ihre Antwort und für diese weiteren Informationen.

    Die Protokolldetails bestätigen meine Vermutung, dass der anti-Myb-Antikörper wohl leider fehlerhaft ist. Zwar ist die Lyse mit Ultraschall vielleicht etwas harsch und kann nach unserer Erfahrung unter Umständen zu unerwünschten Banden führen; allerdings haben Sie ja auch verschiedene Extraktionen ausprobiert, daher ist dieser Punkt sicher nicht der Knackpunkt.

    Es tut mir leid, dass ab45150nicht wie erwartet funktioniert hat. Ich weiß, dass Sie viel Zeit für diese Experimente verwendet haben und möchte Ihnen daher einen kostenlosen Ersatz oder eine Gutschrift anbieten (im Einzelfall können wir die Garantiezeit ausdehnen, und in Ihrem Fall ist diese Ausnahme absolut berechtigt!). Wenn Sie möchten, kann ich Ihnen gerne ein neues Lot von ab45150 zusenden, oder alternativ jeden anderen primären Antikörper Ihrer Wahl. Ich stelle Ihnen auch gerne eine Gutschrift aus, falls Ihnen dies im Moment besser passt.

    Um dies zu arrangieren, benötige ich die ursprüngliche Bestellung. Können Sie dazu mir folgendes bestätigen: Bestellnummer x?

    Vielen Dank für Ihre Kooperation und Hilfsbereitschaft. Ich freue mich auf Ihre Antwort.

    Read More

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