Anti-eNOS (phospho S1177) antibody [EPR20991] (ab215717)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20991] to eNOS (phospho S1177)
- Suitable for: IP, WB, Dot blot
- Reacts with: Mouse, Human
Overview
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Product name
Anti-eNOS (phospho S1177) antibody [EPR20991]
See all eNOS primary antibodies -
Description
Rabbit monoclonal [EPR20991] to eNOS (phospho S1177) -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species IP HumanWB MouseHuman -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HUVEC (+/- VEGF or pervanadate) whole cell lysates; bEND.3 whole cell lysate. IP: HUVEC treated with VEGF whole cell lysate. Dot blot: eNOS (phospho S1177) peptide.
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General notes
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
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
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20991 -
Isotype
IgG -
Research areas
Images
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All lanes : Anti-eNOS (phospho S1177) antibody [EPR20991] (ab215717) at 1/1000 dilution
Lane 1 : bEND.3 (mouse brain endothelioma cell line) whole cell lysate
Lane 2 : bEND.3 (mouse brain endothelioma cell line) treated with alkaline phosphatase for 1 hour, whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Developed using the ECL technique.
Predicted band size: 133 kDa
Observed band size: 140 kDa why is the actual band size different from the predicted?
Exposure time: 2 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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Dot blot analysis of eNOS (phospho S1177) labeled with ab215717 at 1/1000 dilution.
Lane 1: eNOS (phospho S1177) peptide.
Lane 2: eNOS non-phospho peptide.Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution was used as secondary antibody.
Exposure time: 1 minute.
Blocking/Dilution buffer: 5% NFDM/TBST.
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eNOS (phospho S1177) was immunoprecipitated from 0.35 mg of HUVEC (human umbilical vein endothelial cell line) treated with 10 ng/ml VEGF for 5 min, whole cell lysate with ab215717 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab215717 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.
Lane 1: HUVEC treated with VEGF whole cell lysate 10 μg (Input).
Lane 2: ab215717 IP in HUVEC treated with VEGF whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab215717 in HUVEC treated with VEGF whole cell lysate.Exposure time: 30 seconds.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
The 60 kDa band may be a cleaved form of eNOS (PMID: 16195740).
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All lanes : Anti-eNOS (phospho S1177) antibody [EPR20991] (ab215717) at 1/1000 dilution
Lane 1 : HUVEC (human umbilical vein endothelial cell line) whole cell lysate
Lane 2 : HUVEC (human umbilical vein endothelial cell line) treated with 100 uM pervanadate for 10 min, whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 133 kDa
Observed band size: 140 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
The 60 kDa band may be a cleaved form of eNOS (PMID: 16195740).
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All lanes : Anti-eNOS (phospho S1177) antibody [EPR20991] (ab215717) at 1/1000 dilution
Lane 1 : Untreated HUVEC (human umbilical vein endothelial cell line) whole cell lysate
Lane 2 : HUVEC (human umbilical vein endothelial cell line) treated with 10 ng/ml VEGF for 5 min, whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 133 kDa
Observed band size: 140 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
The 60 kDa band may be a cleaved form of eNOS (PMID: 16195740).
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