Overview

  • Product name
    Anti-nNOS (neuronal) (phospho S1417) antibody
    See all nNOS (neuronal) primary antibodies
  • Description
    Rabbit polyclonal to nNOS (neuronal) (phospho S1417)
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-FoFr, WB, IHC-Fr, ICC, IP, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Rabbit, Human
    Predicted to work with: Xenopus laevis, Zebrafish
  • Immunogen

    Synthetic peptide corresponding to Human nNOS (neuronal) aa 1411-1425 (phospho S1417).
    Sequence:

    CNRLRSE(pS)IAFIEESK


    Database link: P29475
    (Peptide available as ab41773)

Properties

Applications

Our Abpromise guarantee covers the use of ab5583 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-FoFr Use at an assay dependent concentration. PubMed: 20881244
WB Use a concentration of 2 µg/ml. Predicted molecular weight: 160 kDa.Can be blocked with Human nNOS (neuronal) (phospho S1417) peptide (ab41773).
IHC-Fr 1/500.
ICC 1/10 - 1/100.
IP Use at an assay dependent concentration.
ICC/IF 1/10 - 1/100.

Target

  • Function
    Produces nitric oxide (NO) which is a messenger molecule with diverse functions throughout the body. In the brain and peripheral nervous system, NO displays many properties of a neurotransmitter. Probably has nitrosylase activity and mediates cysteine S-nitrosylation of cytoplasmic target proteins such SRR.
  • Tissue specificity
    Isoform 1 is ubiquitously expressed: detected in skeletal muscle and brain, also in testis, lung and kidney, and at low levels in heart, adrenal gland and retina. Not detected in the platelets. Isoform 3 is expressed only in testis. Isoform 4 is detected in testis, skeletal muscle, lung, and kidney, at low levels in the brain, but not in the heart and adrenal gland.
  • Sequence similarities
    Belongs to the NOS family.
    Contains 1 FAD-binding FR-type domain.
    Contains 1 flavodoxin-like domain.
    Contains 1 PDZ (DHR) domain.
  • Domain
    The PDZ domain in the N-terminal part of the neuronal isoform participates in protein-protein interaction, and is responsible for targeting nNos to synaptic membranes in muscles. Mediates interaction with VAC14.
  • Post-translational
    modifications
    Ubiquitinated; mediated by STUB1/CHIP in the presence of Hsp70 and Hsp40 (in vitro).
  • Cellular localization
    Cell membrane > sarcolemma. Cell projection > dendritic spine. In skeletal muscle, it is localized beneath the sarcolemma of fast-twitch muscle fiber by associating with the dystrophin glycoprotein complex. In neurons, enriched in dendritic spines.
  • Information by UniProt
  • Database links
  • Alternative names
    • 2310005C01Rik antibody
    • BNOS antibody
    • Constitutive NOS antibody
    • EC 1.14.13.39 antibody
    • IHPS 1 antibody
    • IHPS1 antibody
    • N-NOS antibody
    • NC-NOS antibody
    • neuronal Nitric Oxide Synthase antibody
    • Neuronal NOS antibody
    • Nitric oxide synthase , neuronal, included antibody
    • Nitric oxide synthase 1 (neuronal) antibody
    • Nitric oxide synthase 1 antibody
    • Nitric oxide synthase, brain antibody
    • Nitric oxide synthase, penile neuronal, included antibody
    • NNOS antibody
    • NO antibody
    • NOS 1 antibody
    • NOS antibody
    • NOS type I antibody
    • NOS-I antibody
    • NOS1 antibody
    • NOS1_HUMAN antibody
    • Peptidyl-cysteine S-nitrosylase NOS1 antibody
    see all

Images

  • ab5583 staining nNOS (phospho S1417) in Mouse brain tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with formalin, permeabilized with 0.25% Triton X-100 and blocked with 3% serum for 1 hour at 22°C. Samples were incubated with primary antibody (1/500 in PBS + Triton X-10 + normal donkey serum) for 48 hours at 4°C. A Biotin-conjugated Donkey anti-rabbit IgG polyclonal (1/10000) was used as the secondary antibody.

    See Abreview

  • Immunocytochemistry/Immunofluorescence analysis of nNOS (neuronal) (phospho S1417) (green) showing staining in the cytoplasm and nucleus of SH-SY5Y cells treated with 100 ng/ml nocodazole (left) and untreated SH-SY5Y cells (right). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were incubated with ab5583 in 3% BSA-PBS at a dilution of 1:20 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

References

This product has been referenced in:
  • Wang Z  et al. Quantitative phosphoproteomic analysis of the molecular substrates of sleep need. Nature 558:435-439 (2018). Read more (PubMed: 29899451) »
  • Jean A  et al. Revisiting medial preoptic area plasticity induced in male mice by sexual experience. Sci Rep 7:17846 (2017). IHC-P ; Mouse . Read more (PubMed: 29259324) »
See all 14 Publications for this product

Customer reviews and Q&As

1-10 of 13 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Apteronotus leptorhynchus Tissue lysate - whole (Brain)
Loading amount
50 µg
Specification
Brain
Gel Running Conditions
Reduced Denaturing (4-15%)
Blocking step
Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Jan 31 2013

Answer

Thank you for this information. Based on your protocol I think increasing the amount of primary antibody you use would improve your result. We recommend using 2 ug/ml, which would be 1/500. I would try using this (or even 1/250) overnight at 4 degrees. The GAPDH will be much more highly expressed than the phospho nNOS so I am not surprised that it is showing a good band. Please let me know if you are still having difficulty.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Not Given)
Loading amount
30 µg
Specification
Not Given
Gel Running Conditions
Reduced Denaturing (4-15%)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Sep 04 2012

Answer

I am very pleased to hear you would like to use ab5583 in WB and send us an image of your results.

This code will give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview with the image for WB and include this code in the Additional Comments section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.

For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

The code will be active once the Abreview with the image has been submitted and can be redeemed in one of the following ways:
1) Call to place your order and mention the code to our customer service department;
2) Include the code in your fax order;
3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated. Remember if the product does not work you will be covered by our Abpromise guarantee and eligible for a full refund or replacement.

If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

Read More

Answer

Thank you for your reply.


If you have purchased ab90443 within the last month and are able to submit data in ICC, I am able to issue a code retroactively. However, you will not be able to use this code to obtain ab5583 to test it. You must purchase the antibody to be tested in order to receive a valid code.


I hope this information is helpful. Please let me know and I will issue the code for ab90443 and/or ab5583.

Read More

Answer

Thank you very much for your interest in ab5583 and ab90443.

We do not currently have an image of ab5583 in WB and ab90443 has not been tested in ICC. Therefore, I can offer a discount off a future purchase if you buyab5583 and/or ab90443 now, use it in WB, ICC and submit an image of your results to us in the form of an Abreview. The discount would be to the value of 1 free primary antibody.

It doesn't matter whether the result is positive or negative, we would just really like to receive your feedback. Of course, if the results are negative you will be covered by our Abpromise guarantee and eligible for a refund or replacement.

If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and send an image of ab5583, ab90443 in WB, ICC. I will then send a discount code. This code must be issued before purchasing so please wait for my reply before ordering.

2. Purchase ab5583, ab90443either by phone, fax, or online (www.abcam.com).

3. Use it in WB, ICC.

4. Send us an image of the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for anyprimary antibodyordered and the discount code is valid for 4 months after issue.

Please remember that submission of the Abreview is sufficient for the discount code to become active.

We are always pleased to obtain feedback about our products and any information is greatly appreciated!

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

Read More

Answer

Thank you for your telephone enquiry this afternoon. I am sorry to hear you have had difficulty obtaining satisfactory results from the current vial of this antibody.

I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.

As discussed on the telephone, I would like to reassure you that this antibody is tested and covered by our guarantee for WB and mouse. Before deciding how to proceed, I would like to investigate this particular case further for you, and also obtain some further information for our quality records.

In order to do this, I have enclosed a questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily. I would also appreciate if you are able to provide images, including molecular weight markers, from the vials that have and have not worked, this will be helpful for our investigations and assessment of the results.

In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

Thank you for your time and cooperation. We look forward to receiving the completed questionnaire.

Order Details
Antibody code:
Problem Choose: Non-specific band Multiple bands No signal or weak signal High background
Lot number
Purchase order number or preferably Abcam order number:
General Information Antibody storage conditions (temperature/reconstitution etc)
Description of the problem (high background, wrong band size, more bands, no band etc.)
Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)
Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)
Amount of protein loaded
Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)
Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)
Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
Detection method (ECL, ECLPlus etc.)
Positive and negative controls used (please specify)
Optimization attempts (problem solving) How many times have you tried the Western?
Have you run a "No Primary" control? Yes No
Do you obtain the same results every time? Yes No e.g. are the background bands always in the same place?
What steps have you altered?
Additional Notes:
Image: We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to assess the results.

Read More
Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Apteronotus leptorhynchus Tissue sections (Brain)
Specification
Brain
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.3% Triton X-100
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 24°C

Dr. Ruxandra Sirbulescu

Verified customer

Submitted Nov 24 2011

Application
Western blot
Sample
Human Tissue lysate - whole (Penile Tissue)
Loading amount
60 µg
Specification
Penile Tissue
Gel Running Conditions
Reduced Denaturing (4-15% mini gel)
Blocking step
Starting Block T20 (PBS) as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C

Miss. Gwen Lagoda

Verified customer

Submitted Nov 18 2011

Question
Answer

I am very pleased to hear you would like to accept our offer and test ab16650 and ab5583 in electric fish. This code will give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for electric fish and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews. Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code. Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

Read More

1-10 of 13 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up