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使用敲除细胞株进行验证重组RabMAb

重组Anti-Angiotensin Converting Enzyme 1抗体[EPR2757] (ab75762)

Submit a review Q&A (4)References (14)
Western blot - Anti-Angiotensin Converting Enzyme 1 antibody [EPR2757] (ab75762)
  • ELISA - Anti-Angiotensin Converting Enzyme 1 antibody [EPR2757] (ab75762)
  • Western blot - Anti-Angiotensin Converting Enzyme 1 antibody [EPR2757] (ab75762)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Angiotensin Converting Enzyme 1 antibody [EPR2757] (ab75762)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Angiotensin Converting Enzyme 1 antibody [EPR2757] (ab75762)
  • ELISA - Anti-Angiotensin Converting Enzyme 1 antibody [EPR2757] (ab75762)
  • Anti-Angiotensin Converting Enzyme 1 antibody [EPR2757] (ab75762)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR2757] to Angiotensin Converting Enzyme 1
  • Suitable for: ELISA, WB, IHC-P
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

概述

  • 产品名称

    Anti-Angiotensin Converting Enzyme 1抗体[EPR2757]
    参阅全部 Angiotensin Converting Enzyme 1 一抗

  • 描述

    兔单克隆抗体[EPR2757] to Angiotensin Converting Enzyme 1

  • 宿主

    Rabbit

  • 经测试应用

    适用于: ELISA, WB, IHC-Pmore details
    不适用于: Flow Cyt,ICC/IF or IP

  • 种属反应性

    与反应: Human
    预测可用于: Mouse

  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 阳性对照

    • Fetal kidney, fetal heart and fetal lung lysates; human kidney and spleen tissues.

  • 常规说明

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab75762于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
ELISA
Use a concentration of 0.002 - 2 µg/ml.
WB
1/500 - 1/1000. Detects a band of approximately 195 kDa (predicted molecular weight: 150 kDa).
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
说明
ELISA
Use a concentration of 0.002 - 2 µg/ml.
WB
1/500 - 1/1000. Detects a band of approximately 195 kDa (predicted molecular weight: 150 kDa).
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
应用说明
Is unsuitable for Flow Cyt,ICC/IF or IP.

靶标

  • 功能

    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.

  • 组织特异性

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

  • 疾病相关

    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.

  • 序列相似性

    Belongs to the peptidase M2 family.

  • 翻译后修饰

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

  • 细胞定位

    Secreted and Cell membrane.
  • Information by UniProt

  • 数据库链接

    • 别名

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

    图片

    • Western blot - Anti-Angiotensin Converting Enzyme 1 antibody [EPR2757] (ab75762)
      Western blot - Anti-Angiotensin Converting Enzyme 1 antibody [EPR2757] (ab75762)
      All lanes : Anti-Angiotensin Converting Enzyme 1 antibody [EPR2757] (ab75762) at 1/500 dilution

      Lane 1 : SKNFI cell lysate
      Lane 2 : Ace knockout SKNFI cell lysate
      Lane 3 : HUVEC cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

      Performed under reducing conditions.

      Predicted band size: 150 kDa
      Observed band size: 170 kDa why is the actual band size different from the predicted?



      Anti-Angiotensin Converting Enzyme 1 antibody [EPR2757] staining at 1/500 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab75762 was shown to bind specifically to Angiotensin Converting Enzyme 1. A band was observed at 170 kDa in wild-type SKNFI cell lysates with no signal observed at this size in Ace knockout cell line. To generate this image, wild-type and Ace knockout SKNFI cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

    • ELISA - Anti-Angiotensin Converting Enzyme 1 antibody [EPR2757] (ab75762)
      ELISA - Anti-Angiotensin Converting Enzyme 1 antibody [EPR2757] (ab75762)
      ELISA analysis of Human ACE (angiotensin converting enzyme) (membrane form only) recombinant protein at 250ng/ml with ab75762. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution was used as the secondary antibody.
    • Western blot - Anti-Angiotensin Converting Enzyme 1 antibody [EPR2757] (ab75762)
      Western blot - Anti-Angiotensin Converting Enzyme 1 antibody [EPR2757] (ab75762)
      All lanes : Anti-Angiotensin Converting Enzyme 1 antibody [EPR2757] (ab75762) at 1/1000 dilution

      Lane 1 : fetal kidney lysate
      Lane 2 : fetal heart lysate
      Lane 3 : fetal lung lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : goat anti-rabbit HRP at 1/2000 dilution

      Predicted band size: 150 kDa
      Observed band size: 195 kDa why is the actual band size different from the predicted?

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Angiotensin Converting Enzyme 1 antibody [EPR2757] (ab75762)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Angiotensin Converting Enzyme 1 antibody [EPR2757] (ab75762)

      ab75762 at 1/100 dilution staining Angiotensin Converting Enzyme 1 in human kidney by Immunohistochemistry, Paraffin-embedded tissue.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Angiotensin Converting Enzyme 1 antibody [EPR2757] (ab75762)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Angiotensin Converting Enzyme 1 antibody [EPR2757] (ab75762)

      ab75762 at 1/100 dilution staining Angiotensin Converting Enzyme 1 in human spleen by Immunohistochemistry, Paraffin-embedded tissue. Note positive staining of endothelial cells.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • ELISA - Anti-Angiotensin Converting Enzyme 1 antibody [EPR2757] (ab75762)
      ELISA - Anti-Angiotensin Converting Enzyme 1 antibody [EPR2757] (ab75762)
      ELISA analysis of Human ACE (angiotensin converting enzyme) (membrane form only) recombinant protein at 250 ng/mL with ab75762. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution was used as the secondary antibody.
    • Anti-Angiotensin Converting Enzyme 1 antibody [EPR2757] (ab75762)
      Anti-Angiotensin Converting Enzyme 1 antibody [EPR2757] (ab75762)

    文献 (14)

    发表研究结果有使用 ab75762？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab75762 被引用在 14 文献中.

    • Feng E  et al. Inhibition of HMGB1 Might Enhance the Protective Effect of Taxifolin in Cardiomyocytes via PI3K/AKT Signaling Pathway. Iran J Pharm Res 20:316-332 (2021). PubMed: 34567165
    • Liu R  et al. iTRAQ-based proteomics and in vitro experiments reveals essential roles of ACE and AP-N in the renin-angiotensin system-mediated congenital ureteropelvic junction obstruction. Exp Cell Res 393:112086 (2020). PubMed: 32416091
    • Li SY  et al. Aldosterone from endometrial glands is benefit for human decidualization. Cell Death Dis 11:679 (2020). PubMed: 32826848
    • Fisch S  et al. Localized Antileptin Therapy Prevents Aortic Root Dilatation and Preserves Left Ventricular Systolic Function in a Murine Model of Marfan Syndrome. J Am Heart Assoc 9:e014761 (2020). PubMed: 32378446
    • Xiao L  et al. Wnt/ß-catenin regulates blood pressure and kidney injury in rats. Biochim Biophys Acta Mol Basis Dis 1865:1313-1322 (2019). PubMed: 30710617
    View all Publications for this product

    客户评价及客户问答

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    提交评价 提交问题

    1-4 of 4 Abreviews or Q&A

    Question

    nach längerer Suche kam ich nicht zu dem gleichen Ergebnisse der Enzym Größe. Wie kommen sie auf 138/148 kDa?

    Read More
    Answer

    Wie besprochen habe ich eine kostenlose Ersatzlieferung für Sie in Auftrag gegeben. Sie hat die Referenznummer 1155271 und sollte morgen bei Ihnen ankommen.

    Das Molekulargewicht kann man folgendermaßen errechnen: Öffnen Sie die SwissProt Link auf unserem Datenblatt für das Maus Protein (http://www.uniprot.org/uniprot/P09470), und rollen Sie die Seite einmal bis zum Abschnitt "Sequence annotation (Features)" herunter. Dort finden Sie die Angaben zum "Molecule processing" ,in welchen die Länge und Positionen der einzelnen Protein Ketten beschrieben werde. Wenn Sie die jeweilige Position öffnen, gelangen Sie zur FASTA Sequenz ( in dem Kasten oben links).

    Desweiteren oeffnen Sie den Molekular Gewicht/ pI Rechner unter dem folgenden Link http://web.expasy.org/compute_pi/

    In diesen fügen Sie die FASTA Sequenz ein, und lassen das Gewicht berechnen.

    Ich wünsche Ihnen viel Erfolg für Ihr Projekt. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

    Read More

    Question

    Danke für die schnelle Bearbeitung. Anbei das troubleshooting und drei ausgewählte blots meiner inkubierten Membran.
    Vielen lieben Dank im Vorraus,
    Marion Bisha
    1) Abcam product code ab11737 and ab75762

    2) Abcam order reference number or product batch number
    3) Description of the problem : wrong so signals
    4) Sample preparation:
    Type of sample (whole cell lysates, fraction, recombinant protein…): whole cell lysates, bEND 3 (brain endothelial cells)
    Lysis buffer : Rippa buffer 1ml, PIC 40µl, NaF 10 µl, NaP2O 10 µl, BGP 10 µl, NaV 5 µl
    Boiling for ≥5 min? no 5 min 95°C
    Protein loaded ug/lane 40 Mg / lane
    Positive control ACE rabbit
    Negative control
    5) Percentage of gel: 10%
    Type of membrane : PVDF
    Protein transfer verified
    Blocking agent and concentration; Odyssey Blocking buffer 50% / PBS 50%
    Blocking time : 1 hour
    Blocking temperature room temperature
    6) Primary antibody:
    Concentration or dilution : ab11737= 1:100 and 1:50 and 1:20 ab75762 = 1:1000
    Diluent buffer : Odyssey Blocking buffer 50% / PBS 50% plus 0,1 % Tween
    Incubation time: over night or 4 hours
    Incubation temperature: over night inn 4 °C and 4 hours at room temperature
    7) Secondary antibody: ab11737: anti mouse Odyssey antibody
    Ab75762 anti rabbit Odyssey antibody
    Species:
    Reacts against:
    Concentration or dilution 1:10000
    Diluent buffer : Odyssey Blocking buffer 50% / PBS 50% plus 0,1 % Tween
    Incubation time: 1 hour
    Incubation temperature: room temperature
    Fluorochrome or enzyme conjugate:
    8) Washing after primary and secondary antibodies:
    Buffer: between the antibodies incubation 4 x 5 min with Odyssey Blocking buffer 50% / PBS 50% plus 0,1 % Tween and after the incubation with the second antibody 2 x 5 min with Odyssey Blocking buffer 50% / PBS 50% plus 0,1 % Tween and 2 x 5 min with Odyssey Blocking buffer 50% / PBS 50%
    Number of washes
    9)Detection method
    10) How many times have you run this staining? 5 x
    Do you obtain the same results every time? Jes
    What steps have you altered to try and optimize the use of this antibody?
    Different concentration of the antibodies and different incubation times
    Document attachment: Attaching images of your blot is strongly recommended and can greatly speed up our investigation of your problem.

    Read More
    Answer

    Vielen Dank für Ihren Anruf und dafür, dass Sie sich die Zeit genommen haben, unseren Fragebogen auszufüllen.



    Ich möchte folgende Bemerkungen zu Ihrem Protokoll machen:

    1. Ich denke, Sie detektieren das richtige Protein, da zwei so unterschiedliche Antikörper das gleiche Signal zeigen. Dies kann am veränderten Stoffwechsel der Zellen liegen, die Sie verwenden ( schließlich handelt es sich hier laut ATCC um ein Endothelioma).


    2. Schauen Sie sich bitte einmal das Dokument an, welches ich an diese Email gehängt habe: Dieses Dokument zeigt sehr schön, wie das Gelsystem das Laufverhalten beeinflußen kann.

    3. Eine Abweichung von bis zu 10% von der erwarteten Groesse kann durchaus als normal angesehen werden.

    4. Es gibt zwei Formen dieses Proteins, die lösliche und die zelluläre Form. Die lösliche From ist ca 10 kDa leichter als die zellulare (138 kDa vs 148 kDA).

    5. Da Sie leider mit einem System arbeiten (Odyssey), ist es schwierig Veränderungsvorschläge zu Ihrem Blot zu machen. Es würde mich allerdings interessieren, ob Sie via Chemolumineszenz oder Fluoreszenz detektieren.

    6. Um auszuschließen, dass Sie das falsche Protein detektieren, möchte ich Ihnen ein Maushirn Lysat als Positiv- bzw Vergleichsprobe kostenlos zusenden. Bitte lassen Sie mich wissen, ob Sie damit einverstanden sind.

    Read More

    Question

    mouse, brain lysate; one single band, way to low: 135 kDa.

    Read More
    Answer

    Vielen Dank für Ihren Anruf.

    Es tut mir leid zu hören, dass Sie Probleme mit diesen Antikörpern haben.

    Ich habe unseren Fragebogen als Word-Dokument an diese E-Mail angehängt. Durch das Ausfüllen des Fragebogens erhalten wir alle nötigen Informationen über Ihre Proben und Ihr Protokoll. Sobald Sie dieses Formular an uns zurückgeschickt haben, werden wir uns Ihr Protokoll ansehen und möglichst Veränderungsvorschläge machen, die Ihre Ergebnisse verbessern werden. Falls sich herausstellt, dass der Antikörper nicht so funktioniert, wie auf dem Datenblatt beschrieben und er innerhalb der letzten 180 Tage gekauft wurde, werden wir Ihnen gerne einen Ersatz oder eine Gutschrift schicken.


    Ich freue mich, bald wieder von Ihnen zu hören.

    Read More

    Question

    Can Angiotensin Converting Enzyme 1 antibody [EPR2757] (ab75762) be applicated for detecting  human testicular ACE(tACE or germinal ACE)? Thanks a lot! 我可以Angiotensin Converting Enzyme 1 antibody [EPR2757] (ab75762)抗体检测睾丸特异的ACE（tACE，or germinal ACE）吗？谢谢。 ----------------

    Read More
    Answer

    Thank you for contacting us. The immunogen of ab75762 shares 57% identity with the testis-specific isoform of Angiotensin Converting Enzyme 1. Thus, I would not predict that it would recognize this isoform. 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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