Overview

  • Product name
    Anti-Noxa antibody [114C307]
    See all Noxa primary antibodies
  • Description
    Mouse monoclonal [114C307] to Noxa
  • Host species
    Mouse
  • Tested applications
    Suitable for: Flow Cyt, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Fusion protein containing human Noxa.

  • Positive control
    • WB: RL-7 cell lysate. Flow Cyt: Jurkat cells. IHC-P: Chronic Lymphocytic Leukemia (CLL) xenograft tissue.

Properties

Applications

Our Abpromise guarantee covers the use of ab13654 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
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

IHC-P Use a concentration of 5 µg/ml.

Bouin-fixed.

WB Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 11 kDa.

Target

  • Function
    Promotes activation of caspases and apoptosis. Promotes mitochondrial membrane changes and efflux of apoptogenic proteins from the mitochondria. Contributes to p53/TP53-dependent apoptosis after radiation exposure. Promotes proteasomal degradation of MCL1. Competes with BAK1 for binding to MCL1 and can displace BAK1 from its binding site on MCL1 (By similarity). Competes with BIM/BCL2L11 for binding to MCL1 and can displace BIM/BCL2L11 from its binding site on MCL1.
  • Tissue specificity
    Highly expressed in adult T-cell leukemia cell line.
  • Sequence similarities
    Belongs to the PMAIP1 family.
  • Domain
    The BH3 motif is essential for pro-apoptotic activity.
  • Cellular localization
    Mitochondrion.
  • Information by UniProt
  • Database links
  • Alternative names
    • Adult T cell leukemia derived PMA responsive antibody
    • APR antibody
    • APR_HUMAN antibody
    • ATL-derived antibody
    • Immediate early response protein APR antibody
    • Immediate-early-response protein APR antibody
    • NOXA antibody
    • Phorbol 12 myristate 13 acetate induced protein 1 antibody
    • Phorbol-12-myristate-13-acetate-induced protein 1 antibody
    • PMA induced protein 1 antibody
    • PMA-induced protein 1 antibody
    • PMA-responsive gene antibody
    • Pmaip1 antibody
    • Protein Noxa antibody
    see all

Images

  • Western blot detection of Noxa in RL-7 cell (a follicular lymphoma) lysate. A protein of approximate molecular weight of 11 kDa is detected using ab13654 at 2 µg/ml.

    Western blot detection of Noxa in RL-7 cell (a follicular lymphoma) lysate. A protein of approximate molecular weight of 11 kDa is detected using ab13654 at 2 µg/ml.
  • Overlay histogram showing Jurkat cells stained with ab13654 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab13654, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1](ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 80% methanol/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • Bouin-fixed, paraffin-embedded human tonsil tissue stained for Chronic Lymphocytic Leukemia (CLL) xenograft tissue stained for Noxa using ab13654 at 1/2000 dilution in immunohistochemical analysis.

References

This product has been referenced in:
  • Fitzsimmons L  et al. Coordinated repression of BIM and PUMA by Epstein-Barr virus latent genes maintains the survival of Burkitt lymphoma cells. Cell Death Differ 25:241-254 (2018). WB . Read more (PubMed: 28960205) »
  • Hepp MI  et al. A Trichostatin A (TSA)/Sp1-mediated mechanism for the regulation of SALL2 tumor suppressor in Jurkat T cells. Biochim Biophys Acta N/A:N/A (2018). Human . Read more (PubMed: 29778644) »
See all 44 Publications for this product

Customer reviews and Q&As

1-9 of 9 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (U2OS cells)
Permeabilization
Yes - Tween-20
Specification
U2OS cells
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Acetone

Abcam user community

Verified customer

Submitted Dec 28 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Human Stomach Adenocarcinoma)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH 6.0 Citric Acid
Permeabilization
Yes - Tween-20
Specification
Human Stomach Adenocarcinoma
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Dec 28 2017

Application
Western blot
Loading amount
50 µg
Gel Running Conditions
Reduced Denaturing (18% gel)
Sample
Human Cell lysate - whole cell (Breast)
Specification
Breast
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%

Abcam user community

Verified customer

Submitted Jan 02 2014

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 xxx with ab13687.

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.

Read More

Answer

Thank you for your enquiry regarding ab13654. Noxa is highly expressed in adult T-cell leukemia cell line. Since Jurkat cells are T cell leukemia cell line, it is a good positive control for studying Noxa. Under the References section of Swiss-Prot site, you may be able to find some useful literature: http://www.uniprot.org/uniprot/Q13794 Currently, we have two specific references about this antibody listed on the product datasheet: 1) Sun Ret al. Toll-like receptor 3 (TLR3) induces apoptosis via death receptors and mitochondria by up-regulating the transactivating p63 isoform alpha (TAP63alpha). J Biol Chem 286:15918-28 (2011).Read more (PubMed: 21367858) 2) Morales AAet al. Distribution of Bim determines Mcl-1 dependence or codependence with Bcl-xL/Bcl-2 in Mcl-1-expressing myeloma cells. Blood 118:1329-39 (2011).Read more (PubMed: 21659544) If you need any specific publications regarding the expression on Noxa in Jurkat cells, I would suggest conducting a search on PubMed site: http://www.ncbi.nlm.nih.gov/pubmed/ I hope this helps and if I can assist further, please do not hesitate to contact me.

Read More

Question

I am writing this e-mail to you because we have been experiencing problems with your monoclonal antibody to NOXA (Cat N°. ab13654) for several weeks now.  We have tested this antibody on PVDF and nitrocellulose membrane, with 20, 30 and 40 ug of proteins from Hela cells (in accordance with the abreviews provided on your website), but we always have several aspecific bands in the same range of size as the NOXA protein witch made the identification of the right band really difficult, even in overexpression experiments. Moreover, the detection threshold was really low, since we failed to detect the endogenous protein without a treatment with CisPlatin, despite the loading of as much as 50ug of proteins ( i.e. twice the amounts mentionned in the abreviews). And yet, this really is a problem because we aim to study variations in expression level of the endogenous protein. You will find enclosed a figure composed of two different PVDF membranes incubated with either your antibody (right part) or a monoclonal antibody to Noxa from Calbiochem. The samples used for this experiment are protein extracts coming from Hela cells transfected with a scrambled or Noxa siRNA and treated with Cisplatin to induce expression of endogenous NOXA. As you can see, the signal of the band corresponding to NOXA is really weak compared to the aspecific ones, despite the treatment with cisplatin. In addition the sizes are quite close, which could be a problem since it makes the detection of the endogenous, non-induced protein very delicate (if not impossible)with your antibody. In general we have been very satisfied with the quality of the Abcam products we have purchased and are planning purchase more. Since we have lost a considerable amount of time, money and energy with ab13654 we would like to know if Abcam would be willing to help us with a free antibody or at least discount on our next order.

Read More
Answer

Thank you for contacting us.  I agree that the antibody is not working as expected, and I would be happy to offer you a free of charge replacement, credit, or refund if you have purchased the antibody in the past six months.  It would be very helpful for our quality investigation if you could please provide some additional details about your protocol: 1.)  What was your original order or PO number? 2.)  What blocking agent did you use and at what concentration? 3.)  What was the primary antibody dilution and incubation time? 4.)  What secondary antibody was used, at what concentration, and for how long? If you could please provide this additional information and let me know whether you would prefer a free of charge replacement antibody of your choice, a credit, or a refund, then I will be happy to process your request as soon as possible. 

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Myeloma)
Loading amount
30 µg
Specification
Myeloma
Treatment
1.25 uM RITA for 6 hrs
Gel Running Conditions
Reduced Denaturing (12%)
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C

Manujendra Valued Customer

Verified customer

Submitted Jan 07 2011

Answer

Thank you for your enquiry. Here are some comments and suggestions: 1. You mention using a high dilution of the secondary antibody? What dilution did you use? 2. Further dilution of the primary antibody, perhaps to 0.5ug/ml or 0.25ug/ml, will likely reduce background staining. 3. It might be helpful to see your blot. If you don't mind, please attach a JPEG formatted image of your blot to your reply. Thanks very much. I look forward to hearing from you.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HL60)
Loading amount
20 µg
Specification
HL60
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Jun 26 2007

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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