Recombinant Anti-iNOS antibody [EPR16635] - BSA and Azide free (ab213987)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR16635] to iNOS - BSA and Azide free
- Suitable for: ICC/IF, Indirect ELISA, WB, IP
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-iNOS antibody [EPR16635] - BSA and Azide free
See all iNOS primary antibodies -
Description
Rabbit monoclonal [EPR16635] to iNOS - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, Indirect ELISA, WB, IPmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: RAW 264.7 treated with 0.1 µg/mL LPS for 6 hours, HepG2 treated with 10 µg/mL LPS for 6 hours,whole cell lysates; Human fetal brain lysate; L6 treated with 50 ng/ml IL-1 beta, 20 ng/ml TNF-alpha and 100U/ml IFN-gamma for 24 h, whole cell lysate; ICC/IF: RAW 264.7 cells treated with LPS (0.1 µg/mL), for 6 hours. IP: RAW 264.7 whole cell lysate treated with 1µg/mL LPS for 24h.
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General notes
ab213987 is the carrier-free version of ab178945.
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 <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
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
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. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: 100% PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR16635 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab213987 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 |
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ICC/IF |
1/500.
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Indirect ELISA |
Use at an assay dependent concentration.
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WB |
1/1000. Detects a band of approximately 131 kDa (predicted molecular weight: 131 kDa).
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IP |
1/100.
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Notes |
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ICC/IF
1/500. |
Indirect ELISA
Use at an assay dependent concentration. |
WB
1/1000. Detects a band of approximately 131 kDa (predicted molecular weight: 131 kDa). |
IP
1/100. |
Target
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Function
Produces nitric oxide (NO) which is a messenger molecule with diverse functions throughout the body. In macrophages, NO mediates tumoricidal and bactericidal actions. Also has nitrosylase activity and mediates cysteine S-nitrosylation of cytoplasmic target proteins such COX2. -
Tissue specificity
Expressed in the liver, retina, bone cells and airway epithelial cells of the lung. Not expressed in the platelets. -
Sequence similarities
Belongs to the NOS family.
Contains 1 FAD-binding FR-type domain.
Contains 1 flavodoxin-like domain. - Information by UniProt
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Database links
- Entrez Gene: 4843 Human
- Entrez Gene: 18126 Mouse
- Entrez Gene: 24599 Rat
- Omim: 163730 Human
- SwissProt: P35228 Human
- SwissProt: P29477 Mouse
- SwissProt: Q06518 Rat
- Unigene: 709191 Human
see all -
Alternative names
- HEP-NOS antibody
- Hepatocyte NOS antibody
- HEPNOS antibody
see all
Images
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All lanes : Anti-iNOS antibody [EPR16635] (ab178945) at 1/1000 dilution
Lane 1 : Mouse hippocampus tissue lysate
Lane 2 : Mouse colon tissue lysate
Lane 3 : Mouse colon cancer tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 131 kDa
Observed band size: 131 kDa
Exposure time: 180 secondsThis data was developed using the same antibody clone in a different buffer formulation (ab178945).
Blocking and diluting buffer 5% NFDM/TBST.
iNOS is not normally expressed in the brain, but can be induced in the brain after inflammatory, infectious, or other damages (PMID: 11138926, PMID: 16156895, PMID: 10322315).
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All lanes : Anti-iNOS antibody [EPR16635] (ab178945) at 1/1000 dilution
Lane 1 : Untreated L6 (rat skeletal muscle myoblast) whole cell lysate
Lane 2 : L6 treated with 50 ng/ml IL-1 beta, 20 ng/ml TNF-alpha and 100U/ml IFN-gamma for 24 h, whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 131 kDaThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178945).
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All lanes : Anti-iNOS antibody [EPR16635] (ab178945) at 1/20000 dilution
Lane 1 : Untreated RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
Lane 2 : RAW 264.7 whole cell lysate treated with 0.1 µg/mL LPS for 6 hours
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 131 kDa
Observed band size: 131 kDa
Exposure time: 30 secondsBlocking and dilution buffer: 5% NFDM/TBST.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178945).
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ELISA analysis of House mouse iNOS recombinant protein at 1000 ng/mL with ab178945. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution was used as the secondary antibody.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178945).
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All lanes : Anti-iNOS antibody [EPR16635] (ab178945) at 1/500 dilution
Lane 1 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates
Lane 2 : HepG2 (Human hepatocellular carcinoma epithelial cell) treated with 10ug/ml lipopolysaccharides for 6 hours whole cell lysates
Lane 3 : THP-1 (Human monocytic leukemia monocyte) whole cell lysates
Lane 4 : THP-1 (Human monocytic leukemia monocyte) treated with 100ng/ml lipopolysaccharides for 3 hours whole cell lysates
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/200000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 131 kDa
Exposure time: 3 minutesThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178945).
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iNOS was immunoprecipitated from 1mg of RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate treated with 1 µg/ml LPS for 24h with ab178945 at 1/100 dilution.
Lane 1: RAW 264.7 whole cell lysate treated with 1 µg/ml LPS for 24h,10ug (Input).
Lane 2: ab178945 IP in RAW 264.7 whole cell lysate treated with 1 µg/ml LPS for 24h.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab178945 in RAW 264.7 whole cell lysate treated with 1 µg/ml LPS for 24h.
Western blot was performed from the immunoprecipitate using ab178945 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1500 dilution
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178945).
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Immunocytochemistry/ Immunofluorescence - Anti-iNOS antibody [EPR16635] - BSA and Azide free (ab213987)
Clone EPR16635 (ab213987) has been successfully conjugated by Abcam. This image was generated using Anti-iNOS antibody [EPR16635] (Alexa Fluor® 647). Please refer to ab209027 for protocol details.
Ab209027 staining iNOS in Raw264.7 cells. The lower panel shows cells treated with 1ug/ml Lipopolysaccharides (24 hr). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab209027at 1/100 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
This product also gave a positive signal under the same testing conditions in SW480 cells fixed with 4% formaldehyde (10 min).
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Immunocytochemistry/ Immunofluorescence - Anti-iNOS antibody [EPR16635] - BSA and Azide free (ab213987)
Immunocytochemistry/Immunofluorescence analysis of RAW 264.7 non-treated and LPS treated (0.1 µg/mL) cells labelling iNOS with ab178945 at 1/250. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. The cells were co-stained with ab195889, Alexa Fluor® 594 conjugated anti-alpha tubulin (1/200). Nuclei counterstained with DAPI (blue).
Secondary antibody only controls performed on non-treated and treated cells.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178945).
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (14)
ab213987 has been referenced in 14 publications.
- Yao M et al. Exosomal miR-21 secreted by IL-1ß-primed-mesenchymal stem cells induces macrophage M2 polarization and ameliorates sepsis. Life Sci 264:118658 (2021). PubMed: 33115604
- Wang L et al. The protective role of the miR-25-mediated notch signaling pathway in the memory capacity and brain tissue of mice with central nervous system infections. Am J Transl Res 13:4835-4843 (2021). PubMed: 34150065
- McDowell SAC et al. Neutrophil oxidative stress mediates obesity-associated vascular dysfunction and metastatic transmigration. Nat Cancer 2:545-562 (2021). PubMed: 35122017
- Zhou Z et al. Withaferin A alleviates traumatic brain injury induced secondary brain injury via suppressing apoptosis in endothelia cells and modulating activation in the microglia. Eur J Pharmacol 874:172988 (2020). PubMed: 32032599
- Zhang Z et al. Avicularin Reduces the Expression of Mediators of Inflammation and Oxidative Stress in Bradykinin-Treated MG-63 Human Osteoblastic Osteosarcoma Cells. Med Sci Monit 26:e921957 (2020). PubMed: 32463805