Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Angiotensin Converting Enzyme 1 antibody [EPR22291-247] (ab254222)

Overview

  • Product name

    Anti-Angiotensin Converting Enzyme 1 antibody [EPR22291-247]
    See all Angiotensin Converting Enzyme 1 primary antibodies
  • Description

    Rabbit monoclonal [EPR22291-247] to Angiotensin Converting Enzyme 1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-Fr, WB, IHC-Pmore details
    Unsuitable for: Flow Cyt,ICC/IF or IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human Angiotensin Converting Enzyme 1 aa 600-1250. The exact sequence is proprietary.
    Database link: P12821

  • Positive control

    • WB: bEND.3, HUVEC and HAP1 whole cell lysates; human kidney and lung cell lysates; mouse brain, heart, kidney, spleen and lung tissue lysates; rat brain, heart, liver and spleen tisuue lysates. IHC-P: Human kidney and liver tissue; mouse kidney tissue; rat kidney tissue. IHC-Fr: Mouse kidney tissue; rat kidney tissue.
  • General notes

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab254222 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
IHC-Fr 1/500.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

WB 1/1000. Detects a band of approximately 180 kDa (predicted molecular weight: 150 kDa).
IHC-P 1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF or IP.
  • Target

    • 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
    • Database links

    • Alternative names

      • 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

    Images

    • Immunohistochemical analysis of frozen section of 4% PFA-fixed, 0.2% Triton X-100 permeabilized rat kidney tissue labeling Angiotensin Converting Enzyme 1 with ab254222 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution (green). Positive staining on proximal tubules of kidney (PMID:10504496) is observed. The nuclear counter stain is DAPI (blue).

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

      Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

       

    • All lanes : Anti-Angiotensin Converting Enzyme 1 antibody [EPR22291-247] (ab254222) at 1/1000 dilution

      Lane 1 : HUVEC (human umbilical vein endothelial cell line) whole cell lysate at 20 µg
      Lane 2 : Wild-type HAP1 whole cell lysate at 40 µg
      Lane 3 : Angiotensin Converting Enzyme 1 knockout HAP1 whole cell lysate at 40 µg
      Lane 4 : Human kidney cell lysate at 40 µg
      Lane 5 : Human lung cell lysate at 40 µg

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Developed using the ECL technique.

      Predicted band size: 150 kDa
      Observed band size: 180 kDa
      why is the actual band size different from the predicted?



      Blocking and dilution buffer: 5% NFDM/TBST.

      Exposure times.

      Lanes 1-3: 3 minutes. Lanes 4-5: 5.5 seconds.

      ab254222 was shown to specifically react with Angiotensin Converting Enzyme 1 in wild-type HAP1 cells as signal was lost in Angiotensin Converting Enzyme 1 knockout cells. Wild-type and Angiotensin Converting Enzyme 1 knockout samples were subjected to SDS-PAGE. ab254222 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument using the ECL technique.

      The molecular weight observed, and the expression profile are consistent with what have been described in the literature (PMID: 25495544, 16203874).

       

    • Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling Angiotensin Converting Enzyme 1 with ab254222 at 1/4000 dilution, followed by a Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on proximal tubules of mouse kidney (PMID: 2828286; PMID: 175444) is observed. Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

      The section was incubated with ab254222 for 30 mins at room temperature.
      The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

    • Immunohistochemical analysis of frozen section of 4% PFA-fixed, 0.2% Triton X-100 permeabilized mouse kidney tissue labeling Angiotensin Converting Enzyme 1 with ab254222 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution (green). Positive staining on proximal tubules of kidney (PMID:10504496) is observed. The nuclear counter stain is DAPI (blue).

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

      Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

    • Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling Angiotensin Converting Enzyme 1 with ab254222 at 1/4000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on proximal tubules in mouse kidney (PMID: 2828286; PMID: 175444) is observed. Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

      The section was incubated with ab254222 for 30 mins at room temperature. 
      The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

    • All lanes : Anti-Angiotensin Converting Enzyme 1 antibody [EPR22291-247] (ab254222) at 1/1000 dilution

      Lane 1 : Rat brain tissue lysate
      Lane 2 : Rat heart tissue lysate
      Lane 3 : Rat liver tissue lysate
      Lane 4 : Rat spleen tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 150 kDa
      Observed band size: 180 kDa why is the actual band size different from the predicted?



      Blocking and dilution buffer: 5% NFDM/TBST.

      Exposure times.

      Lane 1: 48 seconds. Lanes 2 & 3:  3 minutes. Lane 4: 48 seconds.

    • Immunohistochemical analysis of paraffin-embedded human liver tissue labeling Angiotensin Converting Enzyme 1 with ab254222 at 1/4000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on blood vessels of human liver (PMID: 175444) is observed. Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

      The section was incubated with ab254222 for 30 mins at room temperature. 
      The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

    • Immunohistochemical analysis of paraffin-embedded human lkidney tissue labeling Angiotensin Converting Enzyme 1 with ab254222 at 1/4000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on proximal tubules of human kidney (PMID: 2828286; PMID: 175444) is observed. Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

      The section was incubated with ab254222 for 30 mins at room temperature. 
      The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

    • All lanes : Anti-Angiotensin Converting Enzyme 1 antibody [EPR22291-247] (ab254222) at 1/1000 dilution

      Lane 1 : Mouse brain tissue lysate
      Lane 2 : Mouse heart tissue lysate
      Lane 3 : Mouse kidney tissue lysate
      Lane 4 : Mouse spleen tissue lysate
      Lane 5 : Mouse lung tissue lysate
      Lane 6 : bEND.3 (mouse brain endothelioma cell line) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 150 kDa
      Observed band size: 180 kDa why is the actual band size different from the predicted?



      Blocking and dilution buffer: 5% NFDM/TBST.

      Exposure times.

      Lanes 1-4: 10 seconds. Lane 5: 5.5 seconds. Lane 6: 48 seconds.

    References

    ab254222 has not yet been referenced specifically in any publications.

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