Anti-nNOS (neuronal) (phospho S847)抗体(ab16650)

概述

  • 产品名称Anti-nNOS (neuronal) (phospho S847)抗体
    参阅全部 nNOS (neuronal) 一抗
  • 描述
    兔多克隆抗体to nNOS (neuronal) (phospho S847)
  • 经测试应用适用于: IHC-Fr, IHC-FoFr, WBmore details
  • 种属反应性
    与反应: Mouse, Rat, Apteronotus leptorhynchus
    预测可用于: Rabbit, Human, Xenopus laevis, Zebrafish
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 800 - 900 of Mouse nNOS (neuronal), phosphorylated at S847.

    (Peptide available as ab16981.)

  • 阳性对照
    • This antibody gave a positive signal in mouse forebrain, mouse spinal cord and mouse cortex tissue lysates. HeLa whole cell lysate was used as a negative control. nNOS is widely expressed in the nervous system: cerebrum, olfactory bulb, hippocampus, midbrain, cerebellum, pons, medulla oblongata, and spinal cord. It is also found in skeletal muscle, spleen, heart, kidney, and liver.

性能

应用

Our Abpromise guarantee covers the use of ab16650 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
IHC-Fr Use at an assay dependent concentration.
IHC-FoFr 1/3000.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 160 kDa (predicted molecular weight: 160 kDa).

靶标

  • 功能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.
  • 组织特异性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.
  • 序列相似性Belongs to the NOS family.
    Contains 1 FAD-binding FR-type domain.
    Contains 1 flavodoxin-like domain.
    Contains 1 PDZ (DHR) domain.
  • 结构域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.
  • 翻译后修饰Ubiquitinated; mediated by STUB1/CHIP in the presence of Hsp70 and Hsp40 (in vitro).
  • 细胞定位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
  • 数据库链接
  • 别名
    • 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

Anti-nNOS (neuronal) (phospho S847) antibody 图像

  • All lanes : Anti-nNOS (neuronal) (phospho S847) antibody (ab16650) at 1 µg/ml

    Lane 1 : Forebrain (Mouse) Tissue Lysate
    Lane 2 : Mouse Cortex Tissue Lysate
    Lane 3 : Spinal Cord (Mouse) Tissue Lysate
    Lane 4 : Forebrain (Mouse) Tissue Lysate with Mouse nNOS (neuronal) (phospho S847) peptide (ab16981) at 1 µg/ml
    Lane 5 : Mouse Cortex Tissue Lysate with Mouse nNOS (neuronal) (phospho S847) peptide (ab16981) at 1 µg/ml
    Lane 6 : Spinal Cord (Mouse) Tissue Lysate with Mouse nNOS (neuronal) (phospho S847) peptide (ab16981) at 1 µg/ml
    Lane 7 : Forebrain (Mouse) Tissue Lysate with nNOS (neuronal) peptide at 1 µg/ml
    Lane 8 : Mouse Cortex Tissue Lysate with nNOS (neuronal) peptide at 1 µg/ml
    Lane 9 : Spinal Cord (Mouse) Tissue Lysate with nNOS (neuronal) peptide at 1 µg/ml

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP)

    Performed under reducing conditions.

    Predicted band size : 160 kDa
    Observed band size : 190 kDa (why is the actual band size different from the predicted?)


    Exposure time : 30 seconds
  • All lanes : Anti-nNOS (neuronal) (phospho S847) antibody (ab16650) at 1 µg/ml

    Lane 1 : Forebrain (Mouse) Tissue Lysate
    Lane 2 : Spinal Cord (Mouse) Tissue Lysate
    Lane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (negative control)

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 160 kDa
    Observed band size : 190 kDa (why is the actual band size different from the predicted?)


    Exposure time : 2 minutes


  • Predicted band size : 160 kDa

    This image is courtesy of Chris Anderson, Wellcome Trust Sanger Institute, United Kingdom

  • Immunostaining using Rabbit polyclonal to nNOS (neuronal) (phospho S847) (ab16650) on rat brain tissue sections (30 micron free floating). ab16650 was used at a dilution of 1/3000 and incubated for 18 hours at RT (in PBS triton 0.3%). Secondary Antibody Goat anti-rabbit alexa Fluor 488 was used at a dilution of 1/1000. The image shows cytoplasmic staining of CNS neurons with ab16650 in naïve rats; the staining being observed in the soma and processes of these neurons. The staining was quenched by pre-incubation with peptide against phospho S847 (ab16981), but not by the control peptide (ab57047) indicating that ab16650 is specific for nNos phosphorylated at S847. Protocol: Rats were perfused-fixed with 4% paraformaldehyde. Tissues were post-fixed overnight in the same fixative and then cryoprotected in 20% sucrose overnight. Following embedding in OCT and freezing, tissues were cut and immunostained using the 'free floating’ technique.
  • ab16650 staining nNOS (neuronal) (phospho S847) in 16 µm thick sections of Apteronotus leptorhynchus by Immunohistochemistry (Frozen sections).
    Tissue was fixed in 2% paraformaldehyde, permeabilized using 0.3% Triton X-100, blocked with 3% sheep serum, 1% BSA, 1% teleostean gelatine in TBS for 1 hour at 24°C and then incubated with ab16650 at a 1/100 dilution for 18 hours at 4°C. The secondary used was an Alexa-Fluor 488 conjugated goat anti-rabbit polyclonal used at a 1/200 dilution.

    See Abreview

Anti-nNOS (neuronal) (phospho S847) antibody (ab16650)参考文献

This product has been referenced in:
  • Niu X  et al. ß3-adrenoreceptor stimulation protects against myocardial infarction injury via eNOS and nNOS activation. PLoS One 9:e98713 (2014). WB ; Mouse . Read more (PubMed: 24911015) »
  • Garzón J  et al. RGSZ2 Binds to the Neural Nitric Oxide Synthase PDZ Domain to Regulate Mu-Opioid Receptor-Mediated Potentiation of the N-Methyl-D-Aspartate Receptor-Calmodulin-Dependent Protein Kinase II Pathway. Antioxid Redox Signal 15:873-87 (2011). WB ; Mouse . Read more (PubMed: 21348811) »

See all 7 Publications for this product

Product Wall

Abcam guarantees this product to work in the 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 14 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

提交于 Sep 28 2012

Thank you for the extra details of your protocol. I think there is nothing wrong with your protocol. I was concerned that you may have been using milk to block the membrane and dilute the antibody, since this can sometimes interfere with detection of p...

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Thank you for bringing this to our attention.

Can you please confirm that you are using ab16650?

I think there are too many bands to identify which is nNOS, but if you send a few details of your western blot protocol, I may be abl...

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Thank you for taking time to complete our questionnaire. I am sorry to hear that this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for...

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Abcam guarantees this product to work in the 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 3% sheep serum, 1% BSA, 1% teleostean gelatine in TBS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 24°C
Username

Dr. Ruxandra Sirbulescu

Verified customer

提交于 Nov 18 2011

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...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - other (Mouse cortex)
Loading amount 15 µg
Specification Mouse cortex
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

提交于 Oct 05 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample Rat Tissue sections (Brain sections (30 um free floating))
Specification Brain sections (30 um free floating)
Fixative Paraformaldehyde
Antigen retrieval step None
Permeabilization No
Username

Dr. Sophie Pezet

Verified customer

提交于 Jan 17 2008

Thank you for the protocol details that you have provided. I would expect you to see a band at approximately 160 kDa using mouse forebrain samples and using ab16650 at a concentration of 1 ug/ml. Since you are not seeing anything at all, even with the ...

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