Anti-CEBP Alpha/CEBPA antibody [15C8] (ab15047)

Submit a review Q&A (5)References (2)

Overview

  • Product name
    Anti-CEBP Alpha/CEBPA antibody [15C8]
    See all CEBP Alpha/CEBPA primary antibodies
  • Description
    Mouse monoclonal [15C8] to CEBP Alpha/CEBPA
  • Host species
    Mouse
  • Specificity
    This antibody may cross react with glutamate--ammonia ligase domain containing 1 protein ((Lengsin) Lens glutamine synthase-like) as it has 8/10 aa identical to the immunogen used to generate ab15047 (this protein is part of the Glutamate synthetase family so its compartmentalization to the cytoplasm) unlike CEDPA which is found in the nucleus.
  • Tested applications
    Suitable for: ICC/IF, WB, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Hamster, Cow, Xenopus laevis, Schizosaccharomyces pombe, Zebrafish, Goldfish, Aspergillus nidulans, Neurospora crassa
  • Immunogen

    Synthetic peptide corresponding to Mouse CEBP Alpha/CEBPA aa 1-14.
    Sequence:

    MESADFYEVEPRPP


    (Peptide available as ab39791)

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • Epitope
    Amino terminal region.
  • Positive control
    • Transfected hu293T cells.
  • General notes

     This product was previously labelled as CEBP Alpha

     

Properties

Applications

Our Abpromise guarantee covers the use of ab15047 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
ICC/IF Use at an assay dependent concentration.
WB Use a concentration of 2 µg/ml. Detects a band of approximately 45 kDa (predicted molecular weight: 45 kDa).Can be blocked with Mouse CEBP Alpha/CEBPA peptide (ab39791).
IP Use at an assay dependent concentration.

Target

  • Function
    C/EBP is a DNA-binding protein that recognizes two different motifs: the CCAAT homology common to many promoters and the enhanced core homology common to many enhancers.
  • Sequence similarities
    Belongs to the bZIP family. C/EBP subfamily.
    Contains 1 bZIP domain.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
    see all
  • Alternative names
    • Apoptotic cysteine protease antibody
    • Apoptotic protease Mch 5 antibody
    • C/EBP alpha antibody
    • C/ebpalpha antibody
    • CAP4 antibody
    • Caspase 8 precursor antibody
    • CBF-A antibody
    • CCAAT Enhancer Binding Protein alpha antibody
    • CCAAT/enhancer binding protein (C/EBP), alpha antibody
    • CCAAT/enhancer-binding protein alpha antibody
    • CEBP antibody
    • CEBP A antibody
    • CEBP alpha antibody
    • Cebpa antibody
    • CEBPA_HUMAN antibody
    • FADD homologous ICE/CED 3 like protease antibody
    • FADD like ICE antibody
    • FLICE antibody
    • ICE like apoptotic protease 5 antibody
    • ICE8 antibody
    • MACH antibody
    • MCH5 antibody
    • MORT1 associated CED 3 homolog antibody
    see all

Images

  • Western blot - Anti-CEBP Alpha/CEBPA antibody [15C8] (ab15047)
    Western blot - Anti-CEBP Alpha/CEBPA antibody [15C8] (ab15047)
    Anti-CEBP Alpha/CEBPA antibody [15C8] (ab15047) at 2 µg/ml + 293 cell lysates transfected with CEBP alpha.

    Predicted band size: 45 kDa

References

This product has been referenced in:
  • Meng X  et al. Heterogeneous nuclear ribonucleoprotein A1 interacts with microRNA-34a to promote chondrogenic differentiation of mesenchymal stem cells. Am J Transl Res 9:1774-1782 (2017). WB ; Mouse . Read more (PubMed: 28469782) »
  • Khanna-Gupta A  et al. Up-regulation of Translation Eukaryotic Initiation Factor 4E in Nucleophosmin 1 Haploinsufficient Cells Results in Changes in CCAAT Enhancer-binding Protein a Activity: IMPLICATIONS IN MYELODYSPLASTIC SYNDROME AND ACUTE MYELOID LEUKEMIA. J Biol Chem 287:32728-37 (2012). WB . Read more (PubMed: 22851180) »
See all 2 Publications for this product

Publishing research using ab15047? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-5 of 5 Abreviews or Q&A

Question

The species of the gene that the plasmid contained is human.   I would be very grateful to you if you could replace cepb alpha mouse ab with rabbit cebp alpha ab40764.       

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Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number **** for ab40764. To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.  

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Question

Thanks for the prompt answer. See my answers below please: 1.What is the species of the transfected CEBP alpha? -Human 293 T cells were transfected with C/ebp alpha plazmid  2.How much protein did you load per lane of the gel? -Approximately 70µg per lane   3.What was the dilution or concentration of the antibody that you incubated with the membrane? - 1:300   I also used your antibody for WB with IP samples and it showed no signal. If you are interested I could send you this WB result.

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Answer

Thank you for sending the additional details. You mentioned that the human cells were transfected with a plasmid; can you please tell me the species of the gene that the plasmid contained? Was this also human, or some other species, for instance mouse/rat, etc? I would like to replace the antibody for you but ab15047 is the only mouse antibody we have for CEBP alpha. Would you be interested in a rabbit monoclonal, for instance ab40761, or ab40764? Click here (or use the following: https://www.abcam.com/index.html?datasheet=40761). Click here (or use the following: https://www.abcam.com/index.html?datasheet=40764).

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Question

Recently I've contacted you because of CEBP Alpha antibody work failure.   I attached our standard protocol for WB procedure. Unfortunately we don't have word version, that's why I'm sending out a PDF format. I was using 10 % separating gel and 4 % stacking gel. The transfer time was 25 min.   I also attached the results of WB with 293 T cells transfected with Cebp alpha. First membrane #1 was incubated with your AB and no signal was obtained. Later I incubated the same membrane with Cebp alpha R and got a signal.   Interestingly, we also bought mouse Cebp alpha from Novus Biologicals (it has the same clone as yours: 15C8) and this AB doesn't work as well. I hope there's a possibility to return the AB or sabstitude this with another clone of Cebp alpha mouse AB. Anyway will be grateful to you for any tips you might give me.    

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Answer

Thank you for sending the protocol and image of your blot. I am sorry to see that you are not obtaining a band that corresponds to CEBP Alpha. I am having some difficulty reading the blot image notations. Can you please answer a few questions, to help me understand the problem? What is the species of the transfected CEBP alpha? How much protein did you load per lane of the gel? What was the dilution or concentration of the antibody that you incubated with the membrane? Finally, what was the antibody that produced the signal in lane four of the image you sent? I see your note, "Later I incubated the same membrane with Cebp alpha R and got a signal", but I am not sure what you mean by Cebp alpha R. Please also tell me your order number for ab15047. I look forward to your reply.

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Question

Many thanks for your e-mail.Following our conversation yesterday,I checked the expasy site and found the protein that you suggests cross reacts with the CEBP alpha antibody.The size ie 57KD fits perfectly with the size of the band that we see on a western blot.Also I have undertaked a 2D blot and obtained a very clean spot in the acidic region of the gel at about pI 5.0.The theorectical pI for the ligase is 5.9.This protein is part of the Glutamate synthetase family so its compartmentalization to the cytoplasm may indeed explain our data.We should be able to pull enough of the protein for mass spec analysis to be absolutely certain.When we do I will let you know,but you should warn your customers of the possable cross reactivities.

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Answer

Many thanks for your feedback, I have added a note on the datasheet following your expert advice and would love to find out if indeed the antibody detected the ligase in your cells. Thank you again for your help, I look forward to hearing from you,

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Question

The customer is detecting the expected 45kDa band in THP1 cells but a 50-55kDA band in B and T cell tumours (human), which is not found in a nuclear fraction but in a cytosolic fraction suggesting the antibody detects another protein or that the protein is abnormally expressed in the cytosol.

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Answer

As discussed earlier I enclose the links to the databases that I was looking at to find out if the antibody could be cross reacting with other proteins. The protein Glutamate--ammonia ligase domain containing 1 protein (Lengsin) Lens glutamine synthase-like) has 8/10 aa identical to the immunogen used to generate ab15047. The information can be found at: http://ca.expasy.org/cgi-bin/niceprot.pl?Q91VC6 I was unable to find any references to a cytoplasmic presence of CEBPA, sorry I couldn't be of more help, Please do not hesitate to contact me again if I can be of further assistance,

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