Overview

  • Product name

  • Detection method

    Colorimetric
  • Sample type

    Cell culture supernatant, Serum, Plasma, Other biological fluids, Tissue Extracts
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    < 5 pg/ml
  • Range

    0.78 ng/ml - 50 ng/ml
  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    Abcam’s Human ACE (CD143) in vitro ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the accurate quantitative measurement of Human ACE (CD143) in cell culture supernatants, serum, plasma (heparin) and saliva.

    A ACE (CD143) specific mouse monoclonal antibody has been precoated onto 96-well plates. Standards and test samples are added to the wells and incubated. A biotinylated detection polyclonal antibody from goat, specific for ACE (CD143) is then added followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex is added and unbound conjugates are washed away with PBS or TBS buffer. TMB is then used to visualize the HRP enzymatic reaction. TMB is catalyzed by HRP to produce a blue color product that changes into yellow after adding acidic stop solution. The density of yellow coloration is directly proportional to the Human ACE (CD143) amount of sample captured in plate.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components Identifier 1 x 96 tests
    ABC Diluent Buffer Blue Cap 1 x 12ml
    Antibody Diluent Buffer Green Cap 1 x 12ml
    Anti-Human ACE Antibody Microplate (12 x 8 wells) 1 unit
    Avidin-Biotin-Peroxidase Complex (ABC) 1 x 130µl
    Biotinylated anti-human ACE antibody 1 x 130µl
    Lyophilized Recombinant Human ACE standard 2 x 50ng
    Plate Seal 1 x 4 units
    Sample Diluent Buffer Green Cap 1 x 30ml
    TMB Color Developing Agent Amber Bottle 1 x 10ml
    TMB Stop Solution Yellow Cap 1 x 10ml
  • Research areas

  • Function

    Converts angiotensin I to angiotensin II by release of the terminal His-Leu, this results in an increase of the vasoconstrictor activity of angiotensin. Also able to inactivate bradykinin, a potent vasodilator. Has also a glycosidase activity which releases GPI-anchored proteins from the membrane by cleaving the mannose linkage in the GPI moiety.
  • Tissue specificity

    Ubiquitously expressed, with highest levels in lung, kidney, heart, gastrointestinal system and prostate. Isoform Testis-specific is expressed in spermatocytes and adult testis.
  • Involvement in disease

    Ischemic stroke (ISCHSTR) [MIM:601367]: A stroke is an acute neurologic event leading to death of neural tissue of the brain and resulting in loss of motor, sensory and/or cognitive function. Ischemic strokes, resulting from vascular occlusion, is considered to be a highly complex disease consisting of a group of heterogeneous disorders with multiple genetic and environmental risk factors. Note=Disease susceptibility is associated with variations affecting the gene represented in this entry.
    Renal tubular dysgenesis (RTD) [MIM:267430]: Autosomal recessive severe disorder of renal tubular development characterized by persistent fetal anuria and perinatal death, probably due to pulmonary hypoplasia from early-onset oligohydramnios (the Potter phenotype). Note=The disease is caused by mutations affecting the gene represented in this entry.
    Microvascular complications of diabetes 3 (MVCD3) [MIM:612624]: Pathological conditions that develop in numerous tissues and organs as a consequence of diabetes mellitus. They include diabetic retinopathy, diabetic nephropathy leading to end-stage renal disease, and diabetic neuropathy. Diabetic retinopathy remains the major cause of new-onset blindness among diabetic adults. It is characterized by vascular permeability and increased tissue ischemia and angiogenesis. Note=Disease susceptibility is associated with variations affecting the gene represented in this entry.
    Intracerebral hemorrhage (ICH) [MIM:614519]: A pathological condition characterized by bleeding into one or both cerebral hemispheres including the basal ganglia and the cerebral cortex. It is often associated with hypertension and craniocerebral trauma. Intracerebral bleeding is a common cause of stroke. Note=Disease susceptibility is associated with variations affecting the gene represented in this entry.
  • Sequence similarities

    Belongs to the peptidase M2 family.
  • Post-translational
    modifications

    Phosphorylated by CK2 on Ser-1299; which allows membrane retention.
  • Cellular localization

    Secreted and Cell membrane.
  • Information by UniProt
  • Alternative names

    • ACE
    • ACE 1
    • ACE T
    • ACE_HUMAN
    • ACE1
    • Angiotensin converting enzyme somatic isoform
    • Angiotensin converting enzyme testis specific isoform
    • Angiotensin I converting enzyme
    • Angiotensin I converting enzyme 1
    • Angiotensin I converting enzyme peptidyl dipeptidase A 1
    • angiotensin I converting enzyme peptidyl-dipeptidase A 1 transcript
    • Angiotensin-converting enzyme
    • Carboxycathepsin
    • CD 143
    • CD143
    • CD143 antigen
    • DCP
    • DCP 1
    • DCP1
    • Dipeptidyl carboxypeptidase 1
    • Dipeptidyl carboxypeptidase I
    • Kininase II
    • MGC26566
    • MVCD3
    • Peptidase P
    • Peptidyl dipeptidase A
    • soluble form
    • Testicular ECA
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab119577 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
Sandwich ELISA Use at an assay dependent concentration.

Images

  • ACE measured in various fluids showing quantity (pg) per mL of tested sample

Protocols

References

This product has been referenced in:

  • Sharma JN  et al. Altered activities of kininase II, an angiotensin converting enzyme, prekallikrein, and nitric oxide in Kuwaiti patients with type 2 diabetes. Int J Immunopathol Pharmacol 28:240-6 (2015). Sandwich ELISA ; Human . Read more (PubMed: 25964383) »
See 1 Publication for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Answer

For ab119577 Human ACE ELISA Kit, this is tested and covered by the Abpromise guarantee for tissue samples. However, I am sorry we have no testing data specifically for muscle samples. Therefore, the sample preparation will need to be individually optimized.

I have copied below a sample preparation protocol which I hope will be helpful:

Tissue Homogenate Preparation:

Rinse tissue with PBS to remove excess blood.
Chop tissue into 1-2 mm pieces on ice in ice-cold buffer, preferably in the presence of protease inhibitors.
Place the tissue in micro-centrifuge tubes and dip them into liquid nitrogen to snap freeze.
Keep samples at -80°C for later use or keep on ice for immediate homogenization.
Prepare the extraction buffer. It can be prepared ahead of time and stored at 4°C.

100 mM Tris, pH 7.4

150 mM NaCl

1 mM EGTA

1 mM EDTA

1% Triton X-100 0.5%

0.5% sodium deoxycholate

Immediately before use, the extraction buffer must be supplemented with the following to generate a complete extraction buffer.
Phosphatase inhibitor cocktail [as directed by manufacturer]
Protease inhibitor cocktail [as directed by manufacturer]
PMSF (Phenyl Methyl Sulfonyl Floride) to 1 mM


For every 5 mg of tissue, add 300 µL of complete extraction buffer to the tube and homogenize.
Rinse the blade of the homogenizer 2X with 300 µL extraction buffer.
Place the sample on a shaker at 4°C for 2 hours.
Centrifuge the sample at 13,000 rpm and 4°C for 20 min.
Assay immediately or aliquot supernatant (soluble protein extract) and hold at -80°C (Avoid freeze/thaw cycles

For suggestions to help with optimization, please see the protocol booklet provided in the link on the online datasheet, page 11.

https://www.abcam.com/ps/products/119/ab119577/documents/ab119577%20ACE%20(CD143)%20Human%20ELISA%20Kit%20v2%20(website).pdf

General Sample information:

The user needs to estimate the concentration of the target protein in the sample and select the correct dilution factor so that the diluted target protein concentration falls near the middle of the linear regime in the standard curve.
Dilute the samples using the provided Sample Diluent Buffer. The following is a guideline for sample dilution. Several trials may be necessary to determine the optimal dilution factor. The sample must be thoroughly mixed with the Diluent buffer before assaying.

High target protein concentration (100 - 1,000 ng/mL). The working dilution is 1:100. i.e. Add 1 μL sample into 99 μL Sample Diluent Buffer


Medium target protein concentration (10 - 100 ng/mL). The working dilution is 1:10. i.e. Add 10 μL sample into 90 μL Sample Diluent Buffer

Low target protein concentration (156 - 10,000 pg/mL). The working dilution is 1:2. i.e. Add 50 μL sample to 50 μL Sample Diluent Buffer

Very Low target protein concentration (≤ 156 pg/mL). No dilution necessary, or the working dilution is 1:2.


 
I hope this will be helpful and am happy to answer any further questions you have.

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Question
Answer

I can now confirm that the antibodies used in the kit are Ace specific, and these should not be cross reacting with ACE2 protein.
I have also done a quick sequence compatibility search between Ace and Ace2 protein, the sequences don’t show high homology therefore the antibodies used in kit should not be cross reacting with Ace2.
http://www.ebi.ac.uk/Tools/services/web/toolresult.ebi?jobId=clustalo-I20160711-115008-0776-23273943-pg

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To detect ACE concentration in urine samples

Good Excellent 5/5 (Ease of Use)
Abreviews
ab119577 - ACE (CD143) Human ELISA Kit allows detection of ACE in urine samples. The urine samples could be used directly without dilution for the experiment. The sample could be prepared in the similar way as mentioned for serum/plasma in the protocol booklet.

Abcam user community

Verified customer

Submitted Apr 12 2016

Answer



I am sorry to confirm that sodium citrate will affect the results generated with this kit. Sodium citrate has chelating properties and will bind to ACE.

I cannot recommend to use samples from sodium citrate tubes.

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Answer

Vielen Dank für Ihre Antwort.

Es tut mir leid, dass wir die Ursache des Problems nicht herausfinden konnten.Wir werdenauch die getesten Proben auf dem Datenblatt updaten und klarer beschreiben.

Ich habe unsere Finanzabteilung beauftragt, eine Gutschrift für Sie auszustellen. Die Referenznummer Ihrer Gutschrift ist xx. Sie können diese Gutschrift auf folgende Weise verwenden:
(1) Gegen die Rechnung dieses Antikörpers verwenden falls diese noch nicht bezahlt wurde
(2) Aufbewahren und gegen eine zukünftige Rechnung verwenden
(3) Wir stellen Ihnen gerne eine komplette Rückerstattung aus, wenn keine offenen Rechnungen mehr ausstehen

Um eine Rückerstattung des Kaufpreises zu erhalten oder um zu bestätigen wie Sie diese Gutschrift benutzen möchten, möchte ich Sie bitten, sich mit unserer Finanzabteilung in Verbindung zu setzen.
Sie können unsere Finanzabteilung per Email unter der Adresse creditcontrol@abcam.com oder telefonisch unter der Nummer 030 896 779 154 erreichen.

Bitte geben Sie die Referenznummer Ihrer Gutschrift immer mit an.

Die Referenznummer ist für Ihre Unterlagen bestimmt. Wir schicken Ihnen die eigentliche Gutschrift per Post oder Email zu. Sie enthält auch die Gutschriftnummer, die mit dem Buchstaben CGB beginnt.

Ich hoffe dass Sie trotz dieser schlechten Erfahrung weiterhin Kunde bei uns bleiben. Bitte zögern Sie nicht sich wieder an uns zu wenden, falls Sie weitere Fragen haben.

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Answer

Vielen Dank für Ihre Antworten.Leider kann ich nicht bestimmen, woher das Problem mit diesem Kit kommen könnte. Ich werde Ihnen deshalb gerne dieses Kit ersetzen - entweder mit einem neuen kostenlosen Kit oder mit einer Gutschrift/Rückerstattung. Es tut mir leid, dass Sie Probleme mit diesemKit hatten. Bitte lassen Sie mich wissen, wie Sie vorgehen möchten.

Ich kann in diesem Moment nicht sagen, ob das Kit welches Sie bekommen haben fehlerhaft ist, oder ob es nicht kompatibel ist mit Urinproben. Unserer Meinung nach, sollte dieses Kit auch mit Urinproben funktionieren, wir haben es jedoch nicht explizit getestet. Vielleicht hat es im Urin eine Substanz, die mit demELISA interferieren könnte - welche dies sein könnte, weiss ich jedoch nicht. Falls Sie als Ersatz ein neues Kit wählen, wäre dies natürlich auch durch unsere Garantie abgedeckt. Sollten Sie wieder keine Resultate mit Urinproben haben, würden wir jedoch denken, dass Urinproben ungeeignet wären. In diesem Falle, könnten wir Ihnen dieses Kit wieder ersetzen, jedoch würden wir Sie auch bitten, uns auch eine Abreview einzureichen, sodass wir dann diese Information mit den anderen Kunden teilen können. Bitte lassen Sie mich wissen, falls Sie Fragen haben.

Ich danke Ihnen für Ihre Kooperation und freue mich wieder von Ihnen zu hören.

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Answer

Vielen Dank für Ihre Anfrage und Ihre Geduld, während ich mit dem Labor in Kontakt war.

Es tut mir leid, dass Sie Probleme mit diesem Kit von uns hatten. In der Tat sind die Daten ziemlich variabel. Sogar die Duplikate sind sehr weit auseinander teilweise. Auch der Hintergrund ist hoch. Wir denken daher, dass die Daten, welche in diesem Experiment gewonnen wurden,nicht zuverlässig sind. Nach der Diskussion mit dem Labor, möchte ich die folgenden Punkte ansprechen. Falls wir das Problem nicht zusammen lösen können, und das Kitfehlerhaft/beschädigt sein sollte,kann ich Ihnen versichern, dass wirIhnen dieses Kit auch ersetzen oder rückerstatten werden.

Hier die Tipps/Angriffspunkte zurVerbesserung der Resultate:

1.)Bei welcher Temperature haben Siedie Inkubation vorgenommen?

2.) Wie wurde das Kit und die Komponenten gelagert?

3.) Können Sie nochmals bestätigen, wie der Waschpuffer benutzt wurde und wievieldavon für jedenWaschgang?Haben Sie zufällig auch den pH gemessen des Waschpuffers?
4.)Wurden die Platten in einem Wasserbad oder Luftinkubator inkubiert?
6.) Könnte die Platte mal ausgetrocknet sein?
Ich freue michauf Ihre Antwort. Vielen Dank für Ihre Kooperation.

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Answer

Thank you for contacting us.

I have been in touch with the development group regarding finding a conversion factor for converting to internation units of ACE. As this product measures total ACE amounts against a known recombinant standard the data from this assay will correspond to the known amount of ACE standard in pg/ml. Although both the development lab and I have attempted to find a conversion factor for this, most databases use nanokatals or other activity measurements to convert to U/L. This would be an inaccurate conversion for the data type presented in this assay. We wish to apologize for any inconvenience this may cause.

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

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Answer

Thank you for contacting us.

EDTA is not recommended as an anticoagulant because of its chelating properties. It may capture Zn ions from the ACE protein and cause changes in the structure that could inhibit antibody binding and lead to reduced or undetectable signal. In theory, you could add some soluble Zn to the samples to neutralize the EDTA and then use heparin as an anticoagulant, but we have not tested this in house.

I hope this helps, please let me know if you need any additional information or assistance.

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Answer

Gracias por contactarnos.

Mi compañera de Servicio al Cliente me ha remitido tu correo.

Nuestros productos se envían en bloques de hielo como medida simplemente preventiva, para mantenerlos a una temperatura constante (salvo que por especificaciones concretas haya que enviarlos a -20C, que en ese caso se envía en hielo seco).

En principio ninguno de los componentes del kit se ha debido degradar por el hecho de no haber llegado a 4C. Los anticuerpos son estables hasta una semana a temperatura ambiente, el estándar esta liofilizado, por lo que no ha podido verse afectado por la temperatura ambiente, y el resto de soluciones son estables también a temperatura ambiente, siempre que sea por un periodo de tiempo corto.

Lo que sí es imprescindible es que una vez recibido el kit, éste se guarde a 4C si se va a usar frecuentemente, o a -20C si se pretende guardarlo durante mayor tiempo.

Así pues, yo os recomiendo que utilicéis el kit normalmente, y en caso de tener el mas mínimo problema os animo a que nos contactéis de inmediato. Todos los productos de Abcam tienen una garantía de 6 meses, por lo que en caso de no funcionar como se especifica en la datasheet éste será reemplazado o reembolsado.

Espero haber resuelto vuestras dudas, en caso contrario, por favor, no dudéis en volver a contactarnos.

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