Overview

  • Product name
    Anti-MMP14 antibody - Hinge region
    See all MMP14 primary antibodies
  • Description
    Rabbit polyclonal to MMP14 - Hinge region
  • Host species
    Rabbit
  • Specificity
    This antibody binds to MMP14, but does not cross react with the other MMP family members (MMP1, MMP2, MMP3, MMP9).
  • Tested applications
    Suitable for: IHC-P, WB, ICC/IF, ELISA, IPmore details
  • Species reactivity
    Reacts with: Mouse, Human, Pig
  • Immunogen

    Synthetic peptide corresponding to Human MMP14.
    (Peptide available as ab41091)

  • Positive control
    • HT1080 cells

Properties

Applications

Our Abpromise guarantee covers the use of ab38971 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
IHC-P Use a concentration of 10 µg/ml.
WB 1/2000 - 1/5000. Predicted molecular weight: 66 kDa. When used against the reduced protein this antibody identifies bands at 65 Kd and 63 Kd (the pro-form and active form) as well as activation/breakdown products.
ICC/IF Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
IP Use at an assay dependent concentration.

Target

  • Function
    Seems to specifically activate progelatinase A. May thus trigger invasion by tumor cells by activating progelatinase A on the tumor cell surface. May be involved in actin cytoskeleton reorganization by cleaving PTK7.
  • Tissue specificity
    Expressed in stromal cells of colon, breast, and head and neck. Expressed in lung tumors.
  • Sequence similarities
    Belongs to the peptidase M10A family.
    Contains 4 hemopexin-like domains.
  • Domain
    The conserved cysteine present in the cysteine-switch motif binds the catalytic zinc ion, thus inhibiting the enzyme. The dissociation of the cysteine from the zinc ion upon the activation-peptide release activates the enzyme.
  • Post-translational
    modifications
    The precursor is cleaved by a furin endopeptidase.
  • Cellular localization
    Membrane. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Information by UniProt
  • Database links
  • Alternative names
    • Matrix metallopeptidase 14 (membrane inserted) antibody
    • Matrix metalloproteinase 14 antibody
    • Matrix metalloproteinase-14 antibody
    • Membrane type 1 matrix metalloproteinase antibody
    • Membrane type 1 metalloprotease antibody
    • Membrane type matrix metalloproteinase 1 antibody
    • Membrane-type matrix metalloproteinase 1 antibody
    • Membrane-type-1 matrix metalloproteinase antibody
    • MMP 14 antibody
    • MMP X1 antibody
    • MMP-14 antibody
    • MMP-X1 antibody
    • Mmp14 antibody
    • MMP14_HUMAN antibody
    • MMPX1 antibody
    • MT MMP 1 antibody
    • MT-MMP 1 antibody
    • MT1 MMP antibody
    • MT1-MMP antibody
    • MT1MMP antibody
    • MTMMP 1 antibody
    • MTMMP1 antibody
    see all

Images

  • All lanes : Anti-MMP14 antibody - Hinge region (ab38971)

    Lane 1 : Cell lysates from human dermal fibroblasts neonatal (no treatment)
    Lane 2 : Cell lysates from human dermal fibroblasts neonatal (treated with II1beta)

    Predicted band size: 66 kDa
    Observed band size: 66 kDa

  • Anti-MMP14 antibody - Hinge region (ab38971) at 1/1000 dilution + HT1080 whole cell lysate at 30 µg

    Secondary
    HRP-conjugated donkey anti-rabbit

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 66 kDa
    Observed band size: 64 kDa
    why is the actual band size different from the predicted?


    Exposure time: 1 minute

    See Abreview

  • All lanes : Anti-MMP14 antibody - Hinge region (ab38971) at 1/5000 dilution

    Lane 1 : Whole cell lysate prepared from HT1080 cells
    Lane 2 : Whole cell lysate prepared from human vascular smooth muscle cells

    Lysates/proteins at 5 µg per lane.

    Secondary
    All lanes : HRP conjugated donkey anti-rabbit IgG at 1/2000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 66 kDa
    Observed band size: 63,65 kDa why is the actual band size different from the predicted?
    Additional bands at: 20 kDa (possible non-specific binding)


    Exposure time: 30 seconds


    Blocked using 5% milk for 1 hour at 20°C.

    See Abreview

  • ab38971 at 10ug/ml staining human HT1080 cells by ICC/IF. The cells were paraformaldhyde fixed and incubated with the antibody for 1 hour. A FITC conjugated donkey anti-rabbit antibody was used as the secondary.

    See Abreview

  • Ab38971 staining Human normal myometrium tissue. Staining is localized to nuclear and cytoplasmic compartment.
    Left panel: with primary antibody at 4 ug/ml. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer citrate pH 6.0 (ab64214) in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS), then incubated with primary antibody for 20 minutes, and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification ma

References

This product has been referenced in:
  • Almalki SG  et al. MMP-2 and MMP-14 Silencing Inhibits VEGFR2 Cleavage and Induces the Differentiation of Porcine Adipose-Derived Mesenchymal Stem Cells to Endothelial Cells. Stem Cells Transl Med 6:1385-1398 (2017). WB ; Pig . Read more (PubMed: 28213979) »
  • Dole NS  et al. Osteocyte-Intrinsic TGF-ß Signaling Regulates Bone Quality through Perilacunar/Canalicular Remodeling. Cell Rep 21:2585-2596 (2017). IHC-P ; Mouse . Read more (PubMed: 29186693) »
See all 28 Publications for this product

Customer reviews and Q&As

1-10 of 18 Abreviews or Q&A

Answer

Thank you for kindly confirming these details as this enables us to closely monitor the quality of our products.
I have been in contact with the lab who have agreed to send us a vial from their new lot free of charge. I am forwarding these details on to our Distributor’s Team who would be happy to organize this.


Thank you for your help and cooperation. Please do not hesitate to contact us if you need anything further.

Read More

Answer

Thank you for taking time to contact us. I am sorry to hear this antibody is not providing satisfactory results.
The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.
When repeating your experiment, I suggest that when preparing your samples, add protease inhibitors in the sample buffer to reduce the digestion of your protein which is what is likely to be causing the band at 14 kDa.
I would also suggest trying to boil your samples in SDS-Page for 5-10 minutes to disrupt any multimers which may be present and could give rise to the higher molecular weight bands.
Having reviewed the protocol details, I believe this product should have given you the correct bands. It appears that you may have received a faulty vial.
I apologize for the inconvenience and am pleased to offer you a free of charge replacement, credit note, or refund in compensation. If you would like a free of charge replacement, unfortunately we do not have any other lot in stock of ab38971. I could recommend ab88618 (www.abcam.com/ab88648) or ab73879 (www.abcam.com/ab73879) or ab51074 (www.abcam.com/ab51074) however none of these antibodies bind to amino acids 285-323 which forms the hinge region.
Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

Read More

Answer

Thanks for the extra information. I have sent you ab58815 (order number ********) so that you can have another try at getting staining in the invadopodia. Since the antibodies that we have previously sent, have been giving staining (just not in the invadopodia), then after sending ab58815, I cannot send anymore replacement antibodies. The antibody (ab58815) is currently out of stock but is due to be available at the end of December, beginning of January. Due to the holidays, I have set the product to ship at the earliest, on January 3rd (if it is stock by then), I just do not want the antibody to arrive and there is the possibility that there will be no one there to receive it. I really hope that this antibody works for you and please let me know if there is anything else I can help you with. Good luck with your experiments.

Read More

Answer

If you do want to test another MMP14 antibody then ab58815 would be a good bet, also ab78738 would be a possible candidate. I have been looking back at the previous MMP14 antibodies you have tried, these are ab38971, ab3644 and ab73879 and all have failed to give you the correct staining that you expected. I was wondering if you had confirmed expression of your protein of interest using other methods, such as western blot, using any of the antibodies that were sent? I can send you ab38971 or ab78738 to try but if that fails to work I will not be able to send you anymore replacement antibodies. The alternative is that I process the refund of the original purchase. I just wanted you to know this before you choose whether to have a replacement antibody sent or not. Please let me know how you would like to cont

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Question
Answer

Thank you for your reply. If the replacement antibody fails to work then I would be happy to send you another MMP14 antibody to try, however if you wish to try a completely different antibody, then I can refund the original purchase and then you can buy whichever antibody you want, whenever. But we cannot credit your account for future purchase, due to the way our accounts system works. Please let me know if there is anything else I can help you with.  

Read More

Answer

Thank you for your reply. If the replacement antibody fails to work as stated in the datasheet, then I would be happy to refund the cost of the original antibody. Please let me know how the alternative methods work out for you and if there is anything else I can help you wi

Read More

Answer

Thank you for contacting us. According to the alignment information that you have sent to me it looks like you should have success with using these antibodies with canine protein. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.  

Read More

Answer

Thank you for your enquiry. We do not have set expiration dates for our products. Most antibodies are stable and can last for anywhere from a few months to several years if stored properly, so we strongly recommend that you follow the storage instructions on the datasheet for the antibody you purchased. These conditions will vary among our antibodies, therefore, it is important to verify the storage conditions for each of our products when you receive them. We guarantee all of our products to work for at least six month from the date of purchase when stored correctly. For more information on antibody storage and stability, please visit our Antibody Storage Guide on our Protocols and Troubleshooting Tips page (https://www.abcam.com/index.html?pageconfig=popular_protocols). I hope this information helps. Please do not hesitate to contact us if you need anything further.

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Question
Answer

The replacement antibody ab73897 should be shipped today, Nov 9th. The fedex tracking number is #******.

Read More

Question
Answer

I have processed your request to receive ab73897 as a replacement for ab3644. I hope that this antibody proves to be successful in your IF staining. Your new order number is *******.

Read More

1-10 of 18 Abreviews or Q&A

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