Anti-TIMP1 antibody (ab61224)

Rabbit polyclonal TIMP1 antibody. Validated in WB, ELISA, IHC, Flow Cyt and tested in Rat, Human. Cited in 18 publication(s). Immunogen corresponding to synthetic peptide.

Overview

  • Product name
  • Description
    Rabbit polyclonal to TIMP1
  • Host species
    Rabbit
  • Specificity
    This antibody has been used successfuly in mouse tissue in a publication, however our WB testing data shows non-reactivity to mouse and we can not guarantee reactivity with mouse.
  • Tested applications
    Suitable for: ELISA, WB, Flow Cyt, IHC-Frmore details
  • Species reactivity
    Reacts with: Rat, Human
    Predicted to work with: Mouse
  • Immunogen

    Synthetic peptide derived from the internal region of human TIMP1.

  • Positive control
    • 293 cell extract, or rat brain lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab61224 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
ELISA 1/10000.
WB 1/500 - 1/1000. Detects a band of approximately 23 kDa (predicted molecular weight: 23 kDa).
Flow Cyt Use at an assay dependent concentration. PubMed: 21133852

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-Fr Use at an assay dependent concentration. PubMed: 21133852

Target

  • Function
    Complexes with metalloproteinases (such as collagenases) and irreversibly inactivates them by binding to their catalytic zinc cofactor. Also mediates erythropoiesis in vitro; but, unlike IL-3, it is species-specific, stimulating the growth and differentiation of only human and murine erythroid progenitors. Known to act on MMP-1, MMP-2, MMP-3, MMP-7, MMP-8, MMP-9, MMP-10, MMP-11, MMP-12, MMP-13 and MMP-16. Does not act on MMP-14.
  • Sequence similarities
    Belongs to the protease inhibitor I35 (TIMP) family.
    Contains 1 NTR domain.
  • Post-translational
    modifications
    The activity of TIMP1 is dependent on the presence of disulfide bonds.
  • Cellular localization
    Secreted.
  • Information by UniProt
  • Database links
  • Alternative names
    • Clgi antibody
    • Collagenase inhibitor antibody
    • Collagenase inhibitor, Human antibody
    • EPA antibody
    • EPO antibody
    • Erythroid Potentiating Activity antibody
    • Erythroid-potentiating activity antibody
    • Fibroblast collagenase inhibitor antibody
    • FLJ90373 antibody
    • HCI antibody
    • Human Collagenase Inhibitor antibody
    • Metalloproteinase inhibitor 1 antibody
    • Metalloproteinase inhibitor 1 precursor antibody
    • OTTHUMP00000023214 antibody
    • TIMP 1 antibody
    • TIMP antibody
    • TIMP metallopeptidase inhibitor 1 antibody
    • TIMP-1 antibody
    • Timp1 antibody
    • TIMP1 protein antibody
    • TIMP1_HUMAN antibody
    • Tissue Inhibitor of Metalloproteinase 1 antibody
    • Tissue inhibitor of metalloproteinases 1 antibody
    • Tissue inhibitor of metalloproteinases antibody
    • Ttissue inhibitor of metalloproteinase 1 erythroid potentiating activity collagenase inhibitor antibody
    see all

Images

  • All lanes : Anti-TIMP1 antibody (ab61224) at 1/500 dilution

    Lane 1 : extracts from 293 cells
    Lane 2 : extracts from 293 cells with the immunizing peptide

    Predicted band size: 23 kDa
    Observed band size: 23 kDa

References

This product has been referenced in:
  • El-Agamy DS  et al. Pristimerin protects against doxorubicin-induced cardiotoxicity and fibrosis through modulation of Nrf2 and MAPK/NF-kB signaling pathways. Cancer Manag Res 11:47-61 (2019). Read more (PubMed: 30588110) »
  • Hu S  et al. Antifertility effectiveness of a novel copper-containing intrauterine device material and its influence on the endometrial environment in rats. Mater Sci Eng C Mater Biol Appl 89:444-455 (2018). Read more (PubMed: 29752117) »
See all 24 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Question
Answer

DISCOUNT CODE: ***
Expiration date: March 9th, 2013
Value: ***

I am very pleased to hear you would like to accept our offer and test ab61224 in mouse. This code will give you *** off your next order before the expiration date. To redeem this offer, please submit an Abreview for mouse and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.
For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: https://www.abcam.com/abtrial.

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Answer

Thank you for contacting Abcam.


Regarding ab1827 the full length protein was used as an immunogen.


Regarding ab61224, it is a peptide from the central region of the protein. The specific sequence is proprietary.


I hope this information is helpful. Please do not hesitate to contact us if you have any additional questions.

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Answer

Thank you for contacting us.

The immunogen for ab61224 correspond to thecentral region of human TIMP1. The full protein sequence can be found by clicking the link; http://www.uniprot.org/uniprot/P01033

The ab1827 is against the whole sequence of human TIMP1. The exact epitope of this antibody hasn't beendetermined yet.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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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. 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

I tried both TIMP antibodies at 1:1000, 1:500, and 1:100. I only got them to work at 1:100 with an overnight exposure. The control antibody I used that detects viral protein was easily detectable. I didn't quantify the amount of protein that I loaded, but I loaded the maximum 15ul volume. I included other samples on the same gel prepared the same way and they were easily detectable via WB using other antibodies. Block: 5% milk, .1% Tween-20 in PBS Wash: 1% milk, .1% Tween-20 in PBS Blocking and AB exposure 1. Trim and mark ladder 2. Wash the membrane in PBS 3. Block 2x 40' RT 4. Wash 2x 5 min in wash 5. Incubate in primary AB diluted in wash O/N 4 degrees 6. Wash 4x 10min in wash 7. Incubate in secondary AB diluted in wash 1 hr RT 8. Wash 4x 10 min in wash 9. Place in PBS 10. Develop I also tried ICC on transfected cells where both antibodies only worked at a 1:10 dilution with the following protocol: 1. Wash in PBS 2. Fix in 4% PFA in PBS 15' RT 3. Wash 3 times in PBS 4. Incubate in blocking solution for 15 min at RT 5. Wash with perm sol. 6. Incubate in perm sol. 15 min at RT 7. Incubate in primary antibody (dilute in perm sol) for 1.5hrs at RT 8. 3x 5 min washes in perm sol. 9. Incubate in secondary antibody for 1.5hrs at RT (dilute in perm. sol.). Cover with foil to protect from light from this point on. 10. 3x 5 min washes in perm sol. 1 1. Incubate in DAPI 1:10,000 in PBS for 10 min at RT 12. Wash with PBS 13. Add vectashield and mount Blocking solution PBS with 3% BSA and .01% sodium azide Permeablization solution PBS with 1% BSA, .1% TritonX-100, .01% sodium azide

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Answer

Thank you for taking the time to send me your protocols relating to this issue. I have been able to review them and agree that the problem here is the product. I would like to offer you free of charge replacements for ab1828 and ab61224. I may also offer you a credit note/refund that may be used against this or any outstanding invoices. As you have mentioned before that you will be using alot of this antibody for future experiments, I thought I might inform you that Abcam will discount orders above 5 units. Please let me know how you would like to procceed and do not hesitate to contact us with further questions or if there are other ways in which we might help you reach your research goals.  

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Answer

Thank you for contacting Abcam in regards to ab1828 and ab61224. I am sorry that you found that these products do not perform as well as expected in your applications.  I would appreciate it if you could provide me with protocol details of the approach that you are using so that I can better determine potential reasons for the reduced performance of these antibodies. I am also very interested in the method of sample preparation that you have used. I would like to offer you assistance if possible for resolving these issues in your experiments. Could you please send me the protocols that you have used for each?   Could you also send the lot numbers for the antibodies and a order number or PO ? Could you also confirm the species of the tissue tested? If we cannot resolve these issues with you I would be happy to offer you a replacement or credit.  

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Question

LOT NUMBER GR41994-1 ORDER NUMBER 39180 DESCRIPTION OF THE PROBLEM Multiple bands SAMPLE lanes 1,2, 3 and 4: human lung cancer cell line (Calu6)whole cell lysate, lanes 5 and 6: normal mammary epithelial cells (MCF10A)whole cell lysate, PRIMARY ANTIBODY anti-TIMP1 (ab61224) diluted 1/200 in PBS-Tween 0,1%, incubation ON at 4 celcius washed 4 times 5 min with PBS-Tw 0,1% DETECTION METHOD Enhanced Luminol (Perkin Elmer) film X-OMAT exposed 2 and 30 min POSITIVE AND NEGATIVE CONTROLS USED We consider MCF10A cells as positive control ANTIBODY STORAGE CONDITIONS aliquot the same day it arrived at the lab and stored at -20 SAMPLE PREPARATION lanes 1,2, 3 and 4 lysing buffer: HEPES 25mM pH7,5, NaCl 150mM, MgCl2 10mM, EDTA 1mM, Trition 0,1%, Glycerol 10%, aprotinin 10microM and sodium vanadate 1mM lanes 5 and 6 lysing buffer:sodium phosphate 20mM, NaCl 150mM, Triton 1%, EDTA 5mM, PMSF 0,2mg/ml, Aprotinin 10microg/ml, leupeptin 10microg/ml, sodium vanadate 0,25mg/ml Sample buffer 2X: Tris 150mM pH 6,8, SDS 1,2%, Glycerol 30%, beta-mercaptoethanol 15%, bromophenol blue 0,0018% Protocole: samples were mixed with sample buffer 2X (1:1) and boiled for 10 min AMOUNT OF PROTEIN LOADED lanes 1,3 and 5 70microg, lanes 2,4 and 6 20microg ELECTROPHORESIS/GEL CONDITIONS 12% resolving gel for denaturing SDS-PAGE and 4% stacking gel TRANSFER AND BLOCKING CONDITIONS wet transfer with Tris-Glycine-SDS buffer with 20% methanol, 900mA for 90min Blocking: PBS-Tween 0,1%- milk 5%, 90min, RT SECONDARY ANTIBODY anti-Rabbit-HRP (Santa Cruz sc-2004) 1/10000 in PBS-Tw 0,1% 90min washed 4 times 5 min with PBS-Tw 0,1% HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? No WHAT STEPS HAVE YOU ALTERED? We changed sample loading buffer. The one before contained less beta-mercaptoethanol and hight molecular weigth appeared on film ADDITIONAL NOTES Is it possible that we detect dimers of TIMP1

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Answer

Thank you for taking time to complete our questionnaire. I am sorry to hear that antibodies ab16123, ab39184 and ab61224 are not providing satisfactory results. The details provided will enable us to investigate these cases and will provide us with vital information for monitoring product quality. Having reviewed these 3 cases, I would like to offer some suggestions to help optimize the results from ab16123, ab39184 and ab61224 : To reduce the background, I recommend to load a maximum of 20µg of samples on the gel. I would suggest to use each primary antibody at a dilution 1/1000 and to dilute each one in the blocking solution. The dilution factor can then be optimized around this value. I would also recommend to try an incubation at room temperature for 2 or 3 hours. Some antibodies bind stonger and more specifically at room temperature. In order to determine the origin of the background I would recommend to run a no primary control in order to see if the background is due to some non-specific binding by the secondary antibody. Last but not least, when testing our antibodies, our lab uses 5% BSA as a blocking reagent, so I recommend switching to this instead of milk. Some antibodies give stronger, more specific signals on blots blocked with BSA instead of milk, so doing this may improve the results you are seeing, and reduce the non-specific bands. An example of the above is the western blot obtained with the Abcam GAPDH antibody ab9385 : www.abcam.com/gapdh-antibody-hrp-loading-control-ab9385.html#GAPDH-Primary-antibodies-ab9385-3.jpg . Should the suggestions not improve the results, please do let me know. In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund. I hope this information is helpful, and I thank you for your cooperation.

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Answer

Thank you for contacting us. We have indeed in our catalog 5 unconjugated anti-interferon alpha 1 antibodies tested and guaranteed to work in Western Blot and Human samples. Please note that, unfortunately, import restrictions make the goat polyclonal antibody ab86316 unavailable to Canadian customers. The remaining 4 products are very similar. However, I would recommend : - the anti-Interferon alpha antibody clone [AE3] reference ab10077 (www.abcam.com/ab10077) or - the anti-Interferon alpha antibody clone [C10F5] reference ab8317 (www.abcam.com/ab8317). These two mouse monoclonal antibodies, protein A purified, are currently in stock, and their unit size is 250µg which makes them cheaper than the other anti-IFNA1. About the other targets, I agree with you, I would recommend - ab39184 for the detection of TIMP3, - ab61224 for TIMP1, - ab95451 (Mouse monoclonal) for Maspin. ab22354 (Rabbit polyclonal) is also a good choice, - ab16123 for CDKN2A/p16INK4a. ab54210 is also well characterised with 2 publications and 3 Abreviews (customer feedbacks). I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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