Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23750-3] to eNOS - BSA and Azide free
  • Suitable for: IHC-P, ICC, IP, Flow Cyt (Intra), WB
  • Reacts with: Human

Overview

  • Product name

    Anti-eNOS antibody [EPR23750-3] - BSA and Azide free
    See all eNOS primary antibodies

  • Description

    Rabbit monoclonal [EPR23750-3] to eNOS - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, ICC, IP, Flow Cyt (Intra), WBmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

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

  • Positive control

    • WB: EA.hy926 and HUVEC whole cell lysates. IHC-P: Human lung carcinoma, kidney, spleen and placenta tissue. ICC: HUVEC cells. Flow Cyt (intra): HeLa cells. IP: EA.hy926 whole cell lysate.

  • General notes

    ab273574 is the carrier-free version of ab252439.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

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

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. 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.

Properties

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)

    This data was developed using ab252439, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling eNOS with ab252439 at 1/500 (1.198 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on endothelial cells in human spleen. The section was incubated with ab252439 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Immunocytochemistry - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
    Immunocytochemistry - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)

    This data was developed using ab252439, the same antibody clone in a different buffer formulation.

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HUVEC cells labelling eNOS with ab252439 at 1/100 (5.99 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in HUVEC cells. Negative control: HeLa (PMID: 19559671). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

  • Flow Cytometry - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
    Flow Cytometry - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)

    This data was developed using ab252439, the same antibody clone in a different buffer formulation.

    Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (human cervix adenocarcinoma epithelial cell, left) /HUVEC (human umbilical vein endothelial cell, right) cells labelling eNOS with ab252439 at 1/50 dilution (1ug)(Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

    A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

    Negative control: Hela (PMID: 19559671).

  • Immunoprecipitation - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
    Immunoprecipitation - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)

    This data was developed using ab252439, the same antibody clone in a different buffer formulation.

    eNOS was immunoprecipitated from 0.35 mg EA.hy926 (human somatic cell hybrid endothelial) whole cell lysate with ab252439 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab252439 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: EA.hy926 (human somatic cell hybrid endothelial) whole cell lysate 10 ug

    Lane 2: ab252439 IP in EA.hy926 (human somatic cell hybrid endothelial) whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab252439 in EA.hy926 (human somatic cell hybrid endothelial) whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 6 seconds.

    IP lysates were unboiled.

  • Western blot - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
    Western blot - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
    Anti-eNOS antibody [EPR23750-3] (ab252439) at 1/1000 dilution + HUVEC (human umbilical vein endothelial cell) whole cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

    Predicted band size: 133 kDa
    Observed band size: 140 kDa why is the actual band size different from the predicted?



    This data was developed using ab252439, the same antibody clone in a different buffer formulation. 

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    HUVEC whole cell lysate in lane 1 is made freshly and used in WB test immediately to minimize protein degradation.

    Exposure time: 6 seconds.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)

    This data was developed using ab252439, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue labeling eNOS with abab252439 at 1/500 (1.198 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on endothelial cells in human lung carcinoma. The section was incubated with ab252439 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Western blot - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
    Western blot - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)

    This data was developed using ab252439, the same antibody clone in a different buffer formulation. 

    Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue labeling eNOS with abab252439 at 1/500 (1.198 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on endothelial cells in human lung carcinoma. The section was incubated with ab252439 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)

    This data was developed using ab252439, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling eNOS with ab252439 at 1/500 (1.198 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining in human placenta. The section was incubated with ab252439 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)

    This data was developed using ab252439, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling eNOS with ab252439 at 1/500 (1.198 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on endothelial cells in human kidney. The section was incubated with ab252439 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
    Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)

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