Overview

  • Product name

    Anti-MT4-MMP antibody [EP1270Y]
    See all MT4-MMP primary antibodies
  • Description

    Rabbit monoclonal [EP1270Y] to MT4-MMP
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, IHC-FoFrmore details
    Unsuitable for: Flow Cyt or IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human MT4-MMP aa 150-250 (internal sequence). The exact sequence is proprietary.

  • Positive control

    • Mouse brain lysate, human lymphoma tissue or HeLa cells.
  • General notes

    Protein previously labeled as MMP17.

    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 ab51075 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. Detects a band of approximately 58 kDa (predicted molecular weight: 67 kDa).
IHC-P Use at an assay dependent concentration.
ICC/IF 1/50 - 1/100.
IHC-FoFr Use at an assay dependent concentration. PubMed: 22022509
  • Application notes
    Is unsuitable for Flow Cyt or IP.
  • Target

    • Function

      Endopeptidase that degrades various components of the extracellular matrix, such as fibrin. May be involved in the activation of membrane-bound precursors of growth factors or inflammatory mediators, such as tumor necrosis factor-alpha. May also be involved in tumoral process. Not obvious if able to proteolytically activate progelatinase A. Does not hydrolyze collagen types I, II, III, IV and V, gelatin, fibronectin, laminin, decorin nor alpha1-antitrypsin.
    • Tissue specificity

      Expressed in brain, leukocytes, colon, ovary testis and breast cancer. Expressed also in many transformed and non-transformed cell types.
    • 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

      Cell membrane. Secreted > extracellular space > extracellular matrix.
    • Information by UniProt
    • Database links

    • Alternative names

      • Matrix metalloproteinase 17 antibody
      • Matrix metalloproteinase 17 membrane inserted antibody
      • Matrix metalloproteinase-17 antibody
      • Membrane type 4 matrix metalloproteinase antibody
      • Membrane type matrix metalloproteinase 4 antibody
      • Membrane-type matrix metalloproteinase 4 antibody
      • Membrane-type-4 matrix metalloproteinase antibody
      • MMP 17 antibody
      • MMP-17 antibody
      • Mmp17 antibody
      • MMP17_HUMAN antibody
      • MT MMP 4 antibody
      • MT MMP4 antibody
      • MT-MMP 4 antibody
      • MT4 MMP antibody
      • MT4-MMP antibody
      • MT4MMP antibody
      • MTMMP 4 antibody
      • MTMMP4 antibody
      see all

    Images

    • Anti-MT4-MMP antibody [EP1270Y] (ab51075) at 1/1000 dilution + Mouse brain lysate at 10 µg

      Secondary
      Goat anti-Rabbit HRP labeled at 1/2000 dilution

      Predicted band size: 67 kDa
      Observed band size: 58 kDa
      why is the actual band size different from the predicted?

    • ab51075 (1:50) staining human MT4-MMP in human lymphoma tissue by immunohistochemistry using paraffin embedded tissue.

    • Immunofluorescent staining of HeLa cells using ab51075 (1:50).
    • Immunohistochemical analysis of rat basolateral amygdala tissue, staining MT4-MMP with ab51075.

      Tissue was permeabilized with 0.5% Triton-X 100 before incubating with primary antibody (1/100) for 48 hours at 4°C. An AlexaFluor®568 goat anti-rabbit IgG was used as the secondary antibody.

    References

    This product has been referenced in:

    • Clemente C  et al. MT4-MMP deficiency increases patrolling monocyte recruitment to early lesions and accelerates atherosclerosis. Nat Commun 9:910 (2018). Read more (PubMed: 29500407) »
    • Blanco MJ  et al. Developmental expression of membrane type 4-matrix metalloproteinase (Mt4-mmp/Mmp17) in the mouse embryo. PLoS One 12:e0184767 (2017). Read more (PubMed: 28926609) »
    See all 10 Publications for this product

    Customer reviews and Q&As

    1-9 of 9 Abreviews or Q&A

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (human cell lines)
    Gel Running Conditions
    Reduced Denaturing (10%)
    Loading amount
    30 µg
    Specification
    human cell lines
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

    Mrs. Monika Schumacher

    Verified customer

    Submitted Feb 25 2015

    Application
    Western blot
    Sample
    Guinea pig Tissue lysate - whole (Liver)
    Gel Running Conditions
    Reduced Denaturing (10 %)
    Loading amount
    30 µg
    Specification
    Liver
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

    Mrs. Monika Schumacher

    Verified customer

    Submitted Jan 23 2015

    Answer

    r these details. I am sorry this vial of antibody has not worked for this customer.

    As requested, we are pleased to arrange an alternative free of charge replacement of ab39028. I can confirm that 1 vial of ab39028 has been added to your order number #####. This has Abcam order reference #####.

    I would like to reassure you that this free of charge replacement vial is also covered by our Abpromise guarantee. Should the customer still be experiencing difficulties with the new vial, or if you have any further questions, please do not hesitate to let me know.

    Thank you for your help and cooperation with this case. Please do not hesitate to contact me if you need anything further.

    Read More

    Answer

    Thank you for your message and for providing this further information.

    I am sorry there are no further suggestions to help improve the results on this occasion. I appreciate the time you have spent on these experiments and would be pleased to arrange a free of charge replacement or credit note in compensation. Alternatively, I am happy to provide a free of charge vial of a different antibody. Please let me know which one you would like to receive.

    I look forward to hearing from you with details of how you would like to proceed.

    Read More

    Question

    Product code: 51075
    Lot number: GR19885-6
    Inquiry: Please see technical enquiry form attached.

    Immunocytochemistry/Immunohistochemistry/ Immunofluorescence
    ANTIBODY DETAILS
    Antibody Code: EP1270Y (ab51075)
    Batch Number: GR19885-6
    GENERAL INFORMATION
    Antibody Storage Conditions (temperature/reconstitution etc)

    Arrived in liquid form from manufacturer

    Description of the Problem (high background, low signal, non-specific staining etc)

    No staining

    Sample (Species/Tissue/Cell Type/Cell Line etc)

    Human placental tissue

    Fixation of Sample (Ethanol/Methanol/Acetone/Paraformaldehyde/Other/Duration)

    We have attempted on both formaldehyde and paraformaldehyde fixed tissue (duration of fixation is usually overnight at 4 degrees

    Antigen Retrieval (Enzymatic method, Heat mediated technique etc)

    We have attempted enzymatic (Trypsin, using Invitrogen’s Digest All Kit), and heating sections in citrate buffer as well as EDTA antigen retrieval buffer for 15-20minutes at high & low power.

    Permeabilization step

    We have tried Triton-X as well as

    Blocking Conditions (Buffer/time period, Blocking agent)

    We use Protein Block (DAKO) for 10minutes

    Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, wash step)

    Abcam Antibody/Rabbit/1%BSA,PBS/1:10, 1:25, 1:50/1-2hr at RT or 37 degrees, overnight at 4 degrees attempted as well/washed with either PBS or PBST after primary antibody
    Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, wash step)

    Invitrogen: Superpicture 3rd gen IHC kit/Universal including rabbit, mouse, rat/ready to use/10-30 minutes/wash with PBS

    Detection Method

    DAB

    Positive and negative controls used (please specify)

    Placental tissue is a positive control for MMP17. Placental tissue has been shown to express MMP17 by both Western Blot and Real-Time PCR methods.

    OPTIMIZATION ATTEMPTS (PROBLEM SOLVING)
    How many times have you tried the IHC?

    30+ sections have been used to optimize the antibody, we have tried everything
    Have you run a “No Primary” control? Yes (Delete one)
    Do you obtain the same results every time? Yes (Delete one)
    What steps have you altered?

    Tried altering incubation time, antibody concentration, antigen retrieval methods (duration and solution)

    Additional Notes

    The antibody does not work at the highest concentration recommended by Abcam (which was 1:50). We have tried even higher concentration without success.

    Read More
    Answer

    Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

    The details you have kindly provided will provide us with vital information for our monitoring of product quality. I would like to reassure you that this antibody is tested and covered by our guarantee for IHC-P and human samples.

    I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. Reviewing the details, I am sorry there not many tips to provide on this occasion to help improve the results. However, I can recommend you may like to consider trying the following options and considerations:

    1. I can suggest to try less time for heat mediated antigen retrieval? This can sometimes require some optimization, and less time is sometimes better than more time. For example, try 5 minutes.

    2. As MMP17 is a cell membrane protein, we would not recommend to permeabilize in Triton. Triton is a strong detergent which can disrupt membrane proteins. It can sometimes significantly affect the staining of membrane targets.

    3. Is the IHC kit working well with other primary antibodies?

    Alternatively, or if these tips do not work, I am pleased to offer you a free of charge replacement or credit note in compensation if the item has been purchased in the 6 month guarantee period. In this case I would appreciate if you are able to confirm the order number.

    Thank you for your cooperation. I look forward to hearing from you with the results of the suggestions details of how you would like to proceed.

    Read More

    Answer

    Thank you for your inquiry.

    The concentration of ab51075 is lot dependent. The lot that we have in stock at the moment and of the last vial you receivedhas a concentration of 0.224 mg/mL.

    I also would like to comment on the seemingly low concentration:

    The IgG produced by rabbits have greater affinity and greater sensitivity than mouse IgG (about 100-1000 fold) and at the same time the total amount of IgG produced by the rabbit hybridomas is generally lower than in mouse hybridomas.

    Many 'veteran' antibody users are generally familiar with the IgG amount provided from mouse antibodies and can judge how many reactions they may get or need to use based on concentration. Unfortunately, this experience is not transferable to rabbit monoclonal antibodies because the concentration scales are generally very different.

    Since the specificity and sensitivity is much higher with rabbit monoclonal antibodies, less IgG is required per reaction.

    I would like to reassure you that while the amount of IgG may not be comparable to mouse monoclonal antibodies, customers are receiving a comparable number of reactions/tests/blots/stainings per tube.

    I hope this information is helpful. Please do not hesitate to contact me with any further questions in this matter and have a good weekend too.

    Read More

    Question
    Answer

    Vielen Dank für Ihren Anruf gestern.

    Ich habe jetzt genauere Information zum Immunogen erhalten:

    Es liegt zwischen den Aminosäuren AA190 und 230 (SwissProt ID Q9ULZ9).

    Ich hoffe, diese Information hilft Ihnen herauszufinden ob die zusätzliche Bande auf dem WB dem "ge-sheddeten" Teilstück des Proteins entspricht.

    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.

    Read More

    Answer

    Thank you for your inquiry. I am happy to confirm that the concentration of lot GR19885 is 0.224 mg/mL. I also would like to comment on the seemingly low concentration: The IgG produced by rabbits have greater affinity and greater sensitivity than mouse IgG (about 100-1000 fold) and at the same time the total amount of IgG produced by the rabbit hybridomas is generally lower than in mouse hybridomas. Many 'veteran' antibody users are generally familiar with the IgG amount provided from mouse antibodies and can judge how many reactions they may get or need to use based on concentration. Unfortunately, this experience is not transferable to rabbit monoclonal antibodies because the concentration scales are generally very different. Since the specificity and sensitivity is much higher with rabbit monoclonal antibodies, less IgG is required per reaction. I would like to reassure you that while the amount of IgG may not be comparable to mouse monoclonal antibodies, customers are receiving a comparable number of reactions/tests/blots/stainings per tube. I hope this information is helpful. Please do not hesitate to contact me with any further questions in this matter.

    Read More

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