Overview

  • Product name
  • Description
    Rabbit polyclonal to MMP7
  • Host species
    Rabbit
  • Specificity
    This antibody specifically recognises MMP7, and does not cross react with MMP1, 2, 3, 8, 9, 10 or 16.
  • Tested applications
    Suitable for: WB, ELISA, IP, IHC-Fr, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Immunogen

    Synthetic peptide corresponding to C-Terminus of MMP7 (Human).

  • Positive control
    • PMA stimulated A431 whole cell lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab5706 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
ELISA
IP
IHC-Fr
ICC/IF
IHC-P
  • Application notes
    ELISA: Use at a concentration of 0.1 - 1.0 µg/ml.
    IHC-Fr: Use at an assay dependent dilution (PMID 18499699).
    IHC-P: Use at a concentration of 2 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
    IP: Use 3-5µg for 106 cells.
    WB: Use at a concentration of 0.5 - 2.0 µg/ml. Detects a band of approximately 28 kDa.


    Not yet tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • Target

    • Function
      Degrades casein, gelatins of types I, III, IV, and V, and fibronectin. Activates procollagenase.
    • Sequence similarities
      Belongs to the peptidase M10A family.
    • 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.
    • Cellular localization
      Secreted > extracellular space > extracellular matrix.
    • Information by UniProt
    • Database links
    • Alternative names
      • Matrilysin antibody
      • Matrin antibody
      • Matrix Metalloproteinase 7 antibody
      • Matrix metalloproteinase-7 antibody
      • MMP 7 antibody
      • MMP-7 antibody
      • MMP7 antibody
      • MMP7_HUMAN antibody
      • MPSL1 antibody
      • PUMP 1 antibody
      • Pump 1 protease antibody
      • Pump-1 protease antibody
      • PUMP1 antibody
      • Uterine matrilysin antibody
      • Uterine metalloproteinase antibody
      see all

    Images

    • Immunohistochemistry analysis of human prostate carcinoma stained with rabbit polyclonal anti-MMP-7 (ab5706).

    • All lanes : Anti-MMP7 antibody (ab5706)

      Lane 1 : Mouse brain lysate
      Lane 2 : Human MMP-7 over-expressing Hek293 cell lysate
    • ab5706 (2µg/ml) staining MMP7 in human kidney cortex using an automated system (DAKO Autostainer Plus). Using this protocol there is staining of the cytoplasm and nuclei within proximal convoluted tubules.
      Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min 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, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.

    References

    This product has been referenced in:
    • Diao B  et al. The role of galectin-3 in the tumorigenesis and progression of pituitary tumors. Oncol Lett 15:4919-4925 (2018). Read more (PubMed: 29545898) »
    • Soler A  et al. Elevated 20-HETE in metabolic syndrome regulates arterial stiffness and systolic hypertension via MMP12 activation. J Mol Cell Cardiol 117:88-99 (2018). WB . Read more (PubMed: 29428638) »
    See all 17 Publications for this product

    Customer reviews and Q&As

    1-9 of 9 Abreviews or Q&A

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (stomach cancer cell)
    Loading amount
    25 µg
    Specification
    stomach cancer cell
    Treatment
    10uM cannabinoid antagonist
    Gel Running Conditions
    Reduced Denaturing (7.5% SDS-PAGE)
    Blocking step
    Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

    Abcam user community

    Verified customer

    Submitted Apr 08 2013

    Answer

    Thanks for your reply.

    If you can provide the PO number or Abcam order reference number associated with the purchase of ab5706 I will be happy to arrange shipment of ab4044 as a replacement.

    Read More

    Answer

    Thanks for sending along the blot image. Here are my thoughts:

    1) Where are the markers and marker labels?;

    2) Has any other antibody been used successfully with these samples?;

    3) It is mentioned 0.5mg protein loaded, Is this the pure MMP7 or colon homogenate?; No more than 0.1mg homogenate should be loaded and 0.5 ug pure protein can be loaded as positive control; loading too much protein will cause proteins to not separate as expected;

    4) 15% gel is too high percentage for MMP7 which is 28kDa, can use 12% for better resolution

    I hope these comments are helpful. Thanks in advance for the additional information.

    Read More

    Question


    Inquiry: Antibody code: ab5706 Batch number: lot:GR1728-7 Antibody storage conditions (temperature/reconstitution etc) When received stored aliquoted and stored immediately at -20 C. Description of the problem (high background, wrong band size, more bands, no band etc.) The antibody seems to be unspecific. Gel shown below shows unspecific binding of the anti-MMP7. Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.) Samples included a positive control of purified recombinant MMP7, and samples from DSS colitis induced mouse colon homogenate, with the negative control being healthy mice. Intended purpose was to use the anti-MMP7 for Western Blot and in the future, immunoprecipitation. Sample preparation (Buffer/Protease inhibitors/Heating sample etc.) Sample was prepared in 0.2%BSA. The western blot was incubated in the primary antibody solution, for one experiment overnight, and for the second trial for one hour at room temperature. The incubation time was not the problem, the same result was obtained. Amount of protein loaded: In this gel, ˜0.5 mg of protein was added, which was lowered in the subsequent experiments. Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.) Reducing PAGE gel, 15%. Transfer and blocking conditions (Buffer/time period, Blocking agent etc.) Transfer conditions were 200 mA for 1hr 15 min. The transfer buffer was made from 1x glycine buffer and 20% methanol. The buffer was prepared in cold milli-Q water. Two blocking solutions were tried in separate experiments, first 3% milk, 100 mM tris pH 7.5, 150 mM NaCl. The second time, the milk was replaced with BSA. Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) The primary antibody is ab5607. Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) The secondary antibody was goat anti rabbit –HRP from Jackson. Detection method (ECL, ECLPlus etc.) ECL was used for detection. Positive and negative controls used (please specify) The positive control here was purifed MMP7. OPTIMIZATION ATTEMPTS (PROBLEM SOLVING) How many times have you tried the Western? Two separate times, two separate secondary antibodies, two separate batches of ECL (from two different labs). Have you run a "No Primary" control? Do you obtain the same results every time? e.g. are the background bands always in the same place? Yes. Both times the gels showed very unspecific binding.

    Read More
    Answer

    Thank you for submitting your completed questionnaire. I am sorry to hear this vial of ab5706 is giving you trouble.

    You mentioned including a western blot in your enquiry but unfortunately it did not make it through. If you could, please re send the blot as an attachment.

    Thanks! I look forward to hearing from you so that we may resolve this as soon as possible.

    Read More

    Answer

    Thank you for confirming this information and for your help and cooperation with this case.

    As requested, I have asked our accounting department to issue you with a credit note. This can then be redeemed against the invoice of a future order.

    Credit ID: #####

    As usual if you have any further questions regarding this credit note, please contact the accounts department by email at creditcontrol@abcam.com. Please refer to the credit ID number in any correspondence with the accounting department.

    I would like to wish the customer good luck with their research. The technical team is always at your service, should you require further expert advice.

    Read More

    Answer

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

    I am sorry to hear the suggestions made have not improved the results on this occasion. I appreciate the time you have spent on these experiments and would be pleased to arrange afree of charge replacementor credit note in compensation.

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

    Read More

    Question

    Dear technical support team:

    This customer has purchase ab5706 (Anti-MMP7 antibody) and has conducted the wb several times with mouse sample. The results show multiple bands, therefore, he wants to ask for your help to modify her experiment step, could you please offer any suggestion to help her?

    I also attached her image in this letter and her experiment step as follow:

    1. Order details:

    Batch number: gr1728-5

    Abcam product code: ab5706

    Antibody storage conditions (temperature/reconstitution etc): -20

    2. Please describe the problem (high background, wrong band size, more bands, no band etc).

    More bands

    3. On what material are you testing the antibody in WB?

    Species: mouse

    What cell line or tissue :aorta

    Cell extract or Nuclear extract: cell extract

    Purified protein or Recombinant protein:



    3. The lysate


    How much protein was loaded: 30micro gram

    What lysis buffer was used: np40



    What protease inhibitors were used: Roche cocktail

    What loading buffer was used: 6x sample dye

    Phosphatase inhibitors Na3VO4

    Did you heat the samples: temperature and time: 95/5 min




    4. Electrophoresis/Gel conditions/ Transfer conditions


    Reducing or non reducing gel: non

    Reducing agent:

    Gel percentage : 10and 15

    Transfer conditions: (Type of membrane, Protein transfer verified): 100v 1hr


    5. Blocking conditions

    Buffer : TBS

    Blocking agent: 5%milk:

    Incubation time: 1hr

    Incubation temperature: RT


    6. Primary Antibody


    Species: R

    Reacts against:


    At what dilution(s) have you tested this antibody: 1:500 or 1:1000

    What dilution buffer was used: 5% milk /TBS

    Incubation time: 4℃ on

    Incubation temperature:

    What washing steps were done:

    7. Secondary Antibody


    Species:

    Reacts against: Rabbit

    At what dilution(s) have you tested this antibody: 1:5000

    Incubation time: 1hr

    Wash steps: 10min with 1% TBST for 3 times

    Fluorochrome or enzyme conjugate:

    Do you know whether the problems you are experiencing come from the secondary?

    8. Detection method
    ECl,

    9. Did you apply positive and negative controls along with the samples? Please specify. No

    10. Optimization attempts

    How many times have you tried the Western? 2 times

    Have you eliminated the possibility that any background bands could be due to the secondary antibody? (Run a “No primary” control): yes

    Do you obtain the same results every time e.g. are background bands always in the same place? yes

    What steps have you altered? Prolong the wash step


    Could you please help this customer to solve the problem?

    Thanks for your kindly help

    Best regards

    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 enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

    I would like to reassure you that ab5706 is tested and covered by our 6 month guarantee for use inWB andmouse samples. In the event that a product is not functioning in the applications cited on the product data sheet, we will be pleased to provide a credit note or free of charge replacement.

    Reviewing this case, I would like to offer some suggestions to help optimize the results from ab5706. I would also appreciate if you can confirm some further details:

    1. Could you confirm the molecular weight of the three very intense bands in the image?Alternatively, I would appreciate ifthe customer would provide a copy of image with molecular weight markers annotated?

    2. I can recommend toreview the protein concentration. The intense bands indicatethe wells may possibly be overloaded. Try loading just 10 - 20 ug of protein per lane.

    3. I can suggest to run the sampleson a reducing gel to ensure the protein runs at the correct molecular weight.

    3. Try a lower concentration of antibody to help reduce any non specific binding. TryDiluting 1:2000.

    4. Is the current vial of secondary antibody working well with other primary antibodies? I can recommend to consider including a no primary control to assess if the secondary is binding non specifically. The concentration of the secondary may need to bereduced to help optimize the results.

    5. I suggest to try BSA rather than milk to block. Changing blocking agent can sometimes help to improve results.

    I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

    Read More
    Application
    Immunocytochemistry
    Sample
    Mouse Cell (Pancreas)
    Specification
    Pancreas
    Blocking step
    Other as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% serum +

    Dr. Howard Crawford

    Verified customer

    Submitted Feb 21 2006

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

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