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  1. Link

    human-timp1-elisa-kit-ab100651.pdf

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Cardiovascular Angiogenesis Adhesion / ECM Matrix Metalloproteinases TIMP
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Human TIMP1 ELISA Kit (ab100651)

  • Datasheet
  • SDS
  • Protocol Booklet
Submit a review Q&A (4)References (6)

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Typical standard curve
  • Typical standard curve

Key features and details

  • Sensitivity: 40 pg/ml
  • Range: 24.69 pg/ml - 18000 pg/ml
  • Sample type: Cell culture supernatant, Plasma, Serum
  • Detection method: Colorimetric
  • Assay type: Sandwich (quantitative)
  • Reacts with: Human

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Overview

  • Product name

    Human TIMP1 ELISA Kit
    See all TIMP1 kits
  • Detection method

    Colorimetric
  • Sample type

    Cell culture supernatant, Serum, Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    < 40 pg/ml
  • Range

    24.69 pg/ml - 18000 pg/ml
  • Recovery

    93 %

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 94.49 84% - 104%
    Serum 92.43 81% - 102%
    Plasma 93.67 83% - 102%
  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    Abcam’s TIMP1 Human ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Human TIMP1 in serum, plasma and cell culture supernatants.


    This assay employs an antibody specific for Human TIMP1 coated on a 96-well plate. Standards and samples are pipetted into the wells and TIMP1 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Human TIMP1 antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of TIMP1 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.


    Get higher sensitivity in only 90 minutes with Human TIMP1 ELISA Kit (ab187394) from our SimpleStep ELISA® range.

  • Notes

    Optimization may be required with urine samples.

  • Platform

    Microplate

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 1 x 96 tests
    20X Wash Buffer 1 x 25ml
    300X HRP-Streptavidin Concentrate 1 x 200µl
    5X Assay Diluent B 1 x 15ml
    Assay Diluent A 1 x 30ml
    Biotinylated anti-Human TIMP1 2 vials
    Recombinant Human TIMP1 Standard (lyophilized) 2 vials
    Stop Solution 1 x 8ml
    TIMP1 Microplate (12 x 8 wells) 1 unit
    TMB One-Step Substrate Reagent 1 x 12ml
  • Research areas

    • Cardiovascular
    • Angiogenesis
    • Adhesion / ECM
    • Matrix Metalloproteinases
    • TIMP
    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Alzheimer's disease
    • Other
    • Immunology
    • Innate Immunity
    • Macrophage / Inflamm.
    • Immunology
    • Innate Immunity
    • Chemokines
    • Beta Chemokine Rec. (CCR)
    • Microbiology
    • Interspecies Interaction
    • Host Virus Interaction
    • Signal Transduction
    • Cytoskeleton / ECM
    • Extracellular Matrix
    • ECM Enzymes
    • TIMP1 / TIMP2
    • Signal Transduction
    • Signaling Pathway
    • G Protein Signaling
    • GPCR
    • Cancer
    • Invasion/microenvironment
    • Angiogenesis
    • ECM enzymes
    • TIMPs
    • Cancer
    • Invasion/microenvironment
    • ECM
    • Extracellular matrix
    • TIMPs
    • Cancer
    • Signal transduction
    • G protein signaling
    • GPCR
    • Cell Biology
    • Proteolysis / Ubiquitin
    • Protease inhibitors
    • Metalloprotease inhibitors
    • TIMPs
    • Cardiovascular
    • Atherosclerosis
    • Vascular Inflammation
    • Leukocyte recruitment
    • Chemokines
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
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    • Immunology
    • Immune System Diseases
    • Antiviral Signaling
    • HIV-related
    • Kits/ Lysates/ Other
    • Kits
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    • ELISA Kits
    • Protease inhibitors ELISA kits
  • 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.
  • Target information above from: UniProt accession P01033 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • Clgi
    • Collagenase inhibitor
    • Collagenase inhibitor, Human
    • EPA
    • EPO
    • Erythroid Potentiating Activity
    • Erythroid-potentiating activity
    • Fibroblast collagenase inhibitor
    • FLJ90373
    • HCI
    • Human Collagenase Inhibitor
    • Metalloproteinase inhibitor 1
    • Metalloproteinase inhibitor 1 precursor
    • OTTHUMP00000023214
    • TIMP
    • TIMP 1
    • TIMP metallopeptidase inhibitor 1
    • TIMP-1
    • Timp1
    • TIMP1 protein
    • TIMP1_HUMAN
    • Tissue Inhibitor of Metalloproteinase 1
    • Tissue inhibitor of metalloproteinases
    • Tissue inhibitor of metalloproteinases 1
    • Ttissue inhibitor of metalloproteinase 1 erythroid potentiating activity collagenase inhibitor
    see all
  • Database links

    • Entrez Gene: 7076 Human
    • Omim: 305370 Human
    • SwissProt: P01033 Human
    • Unigene: 522632 Human

    Associated products

    • SimpleStep ELISA kits

      • Human TIMP1 ELISA Kit (ab187394)

    Images

    • Typical standard curve
      Typical standard curve

      Representative standard curve using ab100651

    • Typical standard curve
      Typical standard curve

      Representative standard curve using ab100651

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (6)

    Publishing research using ab100651? Please let us know so that we can cite the reference in this datasheet.

    ab100651 has been referenced in 6 publications.

    • Lurins J  et al. Impact of several proinflammatory and cell degradation factors in patients with aortic valve stenosis. Exp Ther Med 17:2433-2442 (2019). PubMed: 30906430
    • Wang QW  et al. Anti-influenza A virus activity of rhein through regulating oxidative stress, TLR4, Akt, MAPK, and NF-?B signal pathways. PLoS One 13:e0191793 (2018). PubMed: 29385192
    • D'Souza A  et al. Rationale and design of DUAL study: Doxycycline to Upgrade response in light chain (AL) amyloidosis (DUAL): A phase 2 pilot study of a two-pronged approach of prolonged doxycycline with plasma cell-directed therapy in the treatment of AL amyloidosis. Contemp Clin Trials Commun 8:33-38 (2017). PubMed: 29696194
    • Kapelouzou A  et al. Serum and tissue biomarkers in aortic stenosis. Glob Cardiol Sci Pract 2015:49 (2015). PubMed: 26779524
    • Natrajan MS  et al. Pioglitazone regulates myelin phagocytosis and multiple sclerosis monocytes. Ann Clin Transl Neurol 2:1071-84 (2015). ELISA ; Human . PubMed: 26734659
    • Yang WJ  et al. Matrix metalloproteinase 2 level in human follicular fluid is a reliable marker of human oocyte maturation in in vitro fertilization and intracytoplasmic sperm injection cycles. Reprod Biol Endocrinol 13:102 (2015). PubMed: 26337061

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-4 of 4 Abreviews or Q&A

    Question

    Could you confirm that your Human TIMP-1 ELISA kit (ab100651) is in a form of a 8 wells of 12x strips instead of 1 whole 96 well plate please?

    Thanks

    Read More

    Abcam community

    Verified customer

    Asked on Jan 23 2013

    Answer

    Thank you for contacting us.

    You are correct!, this kit comes with 12 strips of 8 wells.

    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

    Abcam Scientific Support

    Answered on Jan 23 2013

    Question

    Dear technical team,

    Our customer purchased
    ab100586(lot.GR15494),
    ab100662(lot.GR41054),
    ab100606(lot.37486),
    ab100610(lot.GR39199),
    ab100653(lot.GR37277) and
    ab100651(lot.GR37276)
    last year.

    She stored them 4 degrees for 6 months and transferred only standards to -80C.
    And this morning she opened them to use but found out all the 'HRP-Streptavidin Concentrate vials empty.

    She would like to know ifshe can use those kits now and purchase HRP-streptavidin concentrate only.
    But I checked those have different concentrated form, 400x, 300x, 500x or 440x,.

    Please let me know how our customer can use those kits.


    Best regards,
    Youngjin


    ******************************************************
    Technical Support & Marketing Team
    Product Specialist
    T: 82-31-790-4356 C: 82-10-8735-5370
    DAWINBIO Inc.
    601, Blue zone, ITECO. 762, Dukpung-dong,
    Hanam-city, Gyeonggi-do,Korea
    T: 82-1588-9741 F: 82-31-790-4360
    http://www.dawinbio.com/
    ******************************************************

    Read More

    Abcam community

    Verified customer

    Asked on May 21 2012

    Answer

    Thank you for your email. I am sorry for the delay while I was in contact with lab.

    The volume of HRP-Streptavidin is very small and therefore would be difficult to see with the naked eye. The HRP-streptavidin volume sent was 8ul and is very concentrated so it is especially important that the vial is spun down before opening to make sure no solution is adhering to the inside walls or cap of the vial to ensure complete recovery. Please have your researchers perform this preparation and they should be able to use the 8 ul HRP. Below are some additional tips for this component:

    HRP-Streptavidin – comes as a liquid concentrate. Before withdrawing any liquid from the tube, be sure to centrifuge the vial and pipette up and down to mix thoroughly, as precipitation may form in storage. Once diluted, the HRP-strep must be used up that day. Do not store.

    I am also in contact with lab regarding the diluents need to be used for diluting the HRP-Streptavidin vial. I will get back to you with the information soon.

    Please note, the expiration date of kit is 6 month from date of purchase including HRP-Streptavidin. Once the HRP streptavidin is reconstituted it should be used same day.

    Finally, I can suggest using one kit with the provided HRP-Streptavidin, If in case it does not work then you may buy HRP- streptavidin separately. I will confirm the price if you are interested in buying this.

    I hope this information will be helpful. I will contact you with more information soon.

    Read More

    Abcam Scientific Support

    Answered on May 21 2012

    Question

    LOT NUMBER GR37276-1 DESCRIPTION OF THE PROBLEM Problem with standard curve SAMPLE serum PRIMARY ANTIBODY manufacturer DETECTION METHOD as per kit POSITIVE AND NEGATIVE CONTROLS USED n/a ANTIBODY STORAGE CONDITIONS 4 degrees celcius TYPE OF ELISA sandwich COATING WELL manufacturer BLOCKING CONDITIONS n/a SECONDARY ANTIBODY manufacturer HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 1 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? n/a ADDITIONAL NOTES Standard curve did not perform as expected. Protocol was followed exactly. Please see attached results for A450 readings for standards in duplicate. Please advise as I have a large number of Timp-1 assays to run but need a reliable kit. With thanks

    Read More

    Abcam community

    Verified customer

    Asked on Nov 10 2011

    Answer

    Thank you for your inquiry and for forwarding your testing data. There are a few things I noticed after looking at the data: 1) The first thing I noticed was the background is quite high, ~0.5 OD. This high background is really reducing the dynamic range, sensitivity and linearity of the curve so this issue needs to be addressed. Please see tips below for keeping background low: Causes of High Background 1. Too much HRP-Streptavidin may cause background. Briefly centrifuge the vial of HRP-streptavidin concentrate (Item G) and pipette up and down to mix gently before using, since precipitation may form during storage. 2. Too much detection antibody may also contribute to high background. Make sure of correct dilution fold of both the biotinylated antibody (Item F) and HRP-Streptavidin. Both should be diluted with 1x Assay Diluent B. 3. Long incubation times in some steps (i.e. overnight incubation) can cause high background. 4. And finally, the wash steps may introduce background: if the plate is insufficiently washed, or if the wash buffer itself was contaminated. Wash buffer must be removed completely from the wells after each wash. We recommend using an automated plate washer or multi-channel pipettor. Squirt bottles are not advised. If the high background persists after checking all these steps, try reducing the amount of HRP-streptavidin (by increasing the dilution by 1.5 or 2-fold). 2) The second and main issue I see is the non-linear pattern of the standards, specifically from the 18,000pg/ml to the 2,000pg/ml. It would be quite difficult to obtain lower OD response from the 18,000pg/ml and 6,000pg/ml compared to the 2,000pg/ml as the standards are serial diluted. Therefore I don’t think the issue is a standard preparation issue, per se, as there is a noticeable linear pattern from 2,000pg/ml to at least the 666.7pg/ml standard. It seems to be a problem with the two most concentrated points, 6,000 & 18,000 pg/ml. If you omit those points (see lower graph attached), the standard curve is actually okay (r^2 = 0.986). Because the background is so high, it is likely that the highest two standards are reading falsely low (due to precipitate on the bottom of the wells) and that the real values are actually very high and saturated.  Was there any precipitate in the bottom of any of the wells?  If so, signal saturation is probably the issue and can occur if there is too much target protein in the well, causing the enzyme reaction to go too fast, and the color product crashes out of solution.  The most common causes of signal saturation are: 1. Waiting too long before adding stop solution (makes readings artificially high) – make sure to not let the color reaction (TMB incubation) go longer than 30 mins. 2. Doing the color development reaction at high temperature (such as 37 degrees C; also makes readings artificially high) 3. Exposing the plate to light during color development reaction step – light can speed up the reaction and cause falsely high signals. 4. Long incubations – if you incubated overnight, try decreasing to the optional shorter incubation time. 5. Too much HRP-streptavidin – the HRP-streptavidin vial comes highly concentrated so even a small change in loading volume can drastically affect the results ODs. Remember to briefly centrifuge the vial of HRP-streptavidin concentrate (Item G) and pipette up and down to mix gently before using, since precipitation may form during storage. 6. Too much detection antibody - make sure of correct dilution fold of both the biotinylated antibody (Item F) and HRP-Streptavidin. Both should be diluted with 1x Assay Diluent B. 7. Insufficient washings (or contaminated wash buffer) - wash buffer must be removed completely from the wells after each wash. An automated washer or multi-channel pippetor is recommended. Use of a squirt bottle is not advised Because signal saturation and high background are both issues here, I would advise to pay close attention to components that are factors in both, specifically excess HRP-streptavidin and too long TMB incubations. Lastly, you are experiencing some minor high duplicate CV issues which most likely will be corrected once the main issues are addressed but I’ve included some tips for keeping CV low: Causes of High CV 1. Pipette performance – be sure pipettes are calibrated and functioning properly 2. Bubbles in wells – make sure that the plates are not rocked too vigorously during incubations and that there are no bubbles present during reading. 3. Incomplete washing – ensure complete removal of wash buffer after each wash. Also, do not use a squirt bottle to wash the wells – a pipette or a plate washer is recommended. 4. Incomplete reagent mixing (particularly the biotinylated antibody). I hope this information helps. Let me know if the tips do not improve the standard curve readings.

    Read More

    Abcam Scientific Support

    Answered on Nov 10 2011

    Question

    Does the kit detect free TIMP1 only or total TIMP1 (free + complex-bound + truncated)?

    Read More

    Abcam community

    Verified customer

    Asked on Nov 07 2011

    Answer

    Thank you for contacting us. This kit ab100651 will detect free TIMP-1, both pro and active form (thus, total) however it is not known whether it will also detect the bound form (i.e. TIMP-MMP complexes).   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

    Abcam Scientific Support

    Answered on Nov 07 2011

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