Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Angiotensin Converting Enzyme 1 antibody [EPR22291-247] - BSA and Azide free (ab254278)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR22291-247] to Angiotensin Converting Enzyme 1 - BSA and Azide free
  • Suitable for: WB, IHC-P, IHC-Fr, ELISA
  • Knockout validated
  • Reacts with: Mouse, Rat, Human, Recombinant fragment

Overview

  • Product name

    Anti-Angiotensin Converting Enzyme 1 antibody [EPR22291-247] - BSA and Azide free
    See all Angiotensin Converting Enzyme 1 primary antibodies

  • Description

    Rabbit monoclonal [EPR22291-247] to Angiotensin Converting Enzyme 1 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, IHC-Fr, ELISAmore details
    Unsuitable for: Flow Cyt,ICC or IP

  • Species reactivity

    Reacts with: Mouse, Rat, Human, Recombinant fragment

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • 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

    Ab254278 is the carrier-free version of ab254222. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab254278 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Images

  • Immunohistochemistry (Frozen sections) - Anti-Angiotensin Converting Enzyme 1 antibody [EPR22291-247] - BSA and Azide free (ab254278)
    Immunohistochemistry (Frozen sections) - Anti-Angiotensin Converting Enzyme 1 antibody [EPR22291-247] - BSA and Azide free (ab254278)

    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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254222).

  • Immunohistochemistry (Frozen sections) - Anti-Angiotensin Converting Enzyme 1 antibody [EPR22291-247] - BSA and Azide free (ab254278)
    Immunohistochemistry (Frozen sections) - Anti-Angiotensin Converting Enzyme 1 antibody [EPR22291-247] - BSA and Azide free (ab254278)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254222).

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Angiotensin Converting Enzyme 1 antibody [EPR22291-247] - BSA and Azide free (ab254278)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Angiotensin Converting Enzyme 1 antibody [EPR22291-247] - BSA and Azide free (ab254278)

    Immunohistochemical analysis of paraffin-embedded rat 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 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254222).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Angiotensin Converting Enzyme 1 antibody [EPR22291-247] - BSA and Azide free (ab254278)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Angiotensin Converting Enzyme 1 antibody [EPR22291-247] - BSA and Azide free (ab254278)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254222).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Angiotensin Converting Enzyme 1 antibody [EPR22291-247] - BSA and Azide free (ab254278)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Angiotensin Converting Enzyme 1 antibody [EPR22291-247] - BSA and Azide free (ab254278)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254222).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Angiotensin Converting Enzyme 1 antibody [EPR22291-247] - BSA and Azide free (ab254278)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Angiotensin Converting Enzyme 1 antibody [EPR22291-247] - BSA and Azide free (ab254278)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254222).

  • Western blot - Anti-Angiotensin Converting Enzyme 1 antibody [EPR22291-247] - BSA and Azide free (ab254278)
    Western blot - Anti-Angiotensin Converting Enzyme 1 antibody [EPR22291-247] - BSA and Azide free (ab254278)
    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 1/10000 dilution

    Predicted band size: 150 kDa



    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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254222).

     

  • ELISA - Anti-Angiotensin Converting Enzyme 1 antibody [EPR22291-247] - BSA and Azide free (ab254278)
    ELISA - Anti-Angiotensin Converting Enzyme 1 antibody [EPR22291-247] - BSA and Azide free (ab254278)

    This data was developed using ab254222, the same antibody clone in a different buffer formulation.

    ELISA analysis of ACE recombinant protein at 1000 ng/mL with ab254222. 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 [EPR22291-247] - BSA and Azide free (ab254278)
    Anti-Angiotensin Converting Enzyme 1 antibody [EPR22291-247] - BSA and Azide free (ab254278)

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