Anti-nNOS (neuronal) 抗体 (ab6175)

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Immunohistochemistry (Frozen sections) - Anti-nNOS (neuronal) antibody (ab6175)

    Key features and details

    • Sheep polyclonal to nNOS (neuronal)
    • Suitable for: IHC-Fr
    • Reacts with: Rat, Rabbit, Guinea pig
    • Isotype: IgG

    製品の概要

    • 製品名

      Anti-nNOS (neuronal) antibody
      nNOS (neuronal) 一次抗体 製品一覧

    • 製品の詳細

      Sheep polyclonal to nNOS (neuronal)

    • 由来種

      Sheep

    • 特異性

      Reacts with nNOS. Cross-reactivity with iNOS or eNOS not known, but not expected.

    • アプリケーション

      適用あり: IHC-Frmore details

    • 種交差性

      交差種: Rat, Rabbit, Guinea pig

    • 免疫原

      Synthetic peptide:

      RSESIAFIEESKKDADEVFSS

      conjugated to KLH by a Glutaraldehyde linker, corresponding to amino acids 1409-1429 of Rat NOS1 (nNOS).
      Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

    • 特記事項

      This antibody is a high quality immunohistochemical reagent that has been used for studies in both the peripheral and central nervous system. It is also useful for double label studies when combined with rabbit or other primary antibodies.

      Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

      Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

      We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

      In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

      We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

      Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

      Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

    製品の特性

    アプリケーション

    Our Abpromise guarantee covers the use of ab6175 in the following tested applications.

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

    アプリケーション Abreviews 特記事項
    IHC-Fr 1/100 - 1/2000.

    ターゲット情報

    • 機能

      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.

    • 組織特異性

      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.

    • 配列類似性

      Belongs to the NOS family.
      Contains 1 FAD-binding FR-type domain.
      Contains 1 flavodoxin-like domain.
      Contains 1 PDZ (DHR) domain.

    • ドメイン

      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.

    • 翻訳後修飾

      Ubiquitinated; mediated by STUB1/CHIP in the presence of Hsp70 and Hsp40 (in vitro).

    • 細胞内局在

      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

    • 参照データベース

      • 別名

        • 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

      画像

      • Immunohistochemistry (Frozen sections) - Anti-nNOS (neuronal) antibody (ab6175)
        Immunohistochemistry (Frozen sections) - Anti-nNOS (neuronal) antibody (ab6175)

        Rat brain stem tissue staining for nNOS with ab6175 (1:2000) (perfusion fixed frozen sections).

      参考文献 (5)

      ab6175 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

      ab6175 は 5 報の論文で使用されています。

      • Reuss S  et al. Neuroglobin Expression in the Mammalian Auditory System. Mol Neurobiol 53:1461-1477 (2016). IHC-FoFr ; Mouse . PubMed: 25636685
      • Stenkamp-Strahm CM  et al. Prolonged high fat diet ingestion, obesity, and type 2 diabetes symptoms correlate with phenotypic plasticity in myenteric neurons and nerve damage in the mouse duodenum. Cell Tissue Res 361:411-26 (2015). PubMed: 25722087
      • Colucci M  et al. Intestinal dysmotility and enteric neurochemical changes in a Parkinson's disease rat model. Auton Neurosci 169:77-86 (2012). PubMed: 22608184
      • Buchwalow IB  et al. Nitric oxide synthase in muscular dystrophies: a re-evaluation. Acta Neuropathol (Berl) 111:579-88 (2006). PubMed: 16718354
      • Buchwalow IB  et al. Compartmentalization of NO signaling cascade in skeletal muscles. Biochem Biophys Res Commun 330:615-21 (2005). PubMed: 15796928

      レビューと Q&A

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      1-9 of 9 Abreviews or Q&A

      Question

      with regard to the antibodies that have been tested and guaranteed (especially for ab72428), what rabbit tissues were used, and what was the localization of the staining? using the same tissue will help to at least confirm the reported reactivity. Aside from your own in-house validation, do you have record of publications that use your antibodies? specifically, are there any that use the antibodies that you've suggested in rabbit tissue IHC-P?

      specifically regarding ab72428, the antigen for the antibody is a pentamer from the nNOS c-terminus. with such a short sequence, is there concern for cross-reactivity, especially to iNOS or eNOS, forexample?

      thanks for suggesting some other alternative antibodies. Unfortunately ab79051 and ab95436 use a similar antigen to ab6175 and ab1376 (1400-1429 aa region in nNOS), so I would not consider those. ab106417, the last one you suggested, is a rabbit polyclonal. is this an issue when applying the antibody to rabbit tissue?

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      Answer

      Thank you for your reply.

      As for ab72428, I will need to check with the lab regarding tested tissue,localization of staining and potential cross-reactivity with eNOS and iNOS. I will letyou know what I find out. Thank you for your patience.

      As for references, we list them on the online datasheet in a tab called "Specific references" if we are aware of any. Also, customer reviews can contain testing data on new species and those would be listed in the tab "Abreviews" on the online datasheet.

      For ab72428:
      References: https://www.abcam.com/nNOS-neuronal-antibody-C-terminal-ab72428-references.html
      Abreviews: wehave not yet received Abreviews from other customers for this antibody.

      As for ab106417, a rabbit polyclonal, yes, high background due to crossreactivity of the secondary antibody (anti-rabbit) with the endogenous rabbit IgG (just as mouse-on-mouse staining)could potentially pose a problem, but there are ways to get around this.
      Here is our mouse-on-mouse staining advice:
      https://www.abcam.com/index.html?pageconfig=resource&rid=11465
      which can be adapted to rabbit-on-rabbit staining as well.

      I hope this information is of some help so far. I will be in touch with more details soon.
      In the meantime, please do not hesitate to contact us if you need any more advice or information.

      Use our products? Submit an Abreview. Earn rewards!
      https://www.abcam.com/abreviews

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      Question

      Inquiry: I am interested in purchasing an nNOS antibody for use in immunohistochemistry/immunofluorescence in rabbit paraffin sections. A number of your antibodies in your catalog state they have reactivity with rabbit, and have been documented for use in IHC-P. In particular, I am interested in ab3511 and ab72428. ab6175 and ab1376 meet the criteria, but use a similar antigen to another antibody that I tested and didn't work.
      Have any antibodies in your catalog been documented to work specifically for IHC-P in rabbit tissue? which would you recommend that I try?

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      Answer

      Thank you for contacting us.

      We have 3 antibodies that were tested and are guaranteed (https://www.abcam.com/Abpromise) for IHC-P and rabbit samples: ab1376,ab6175 and ab72428. I would recommend trying one of thosethat were tested as they are covered under our guarantee.
      We do not compare antibodies side by side, thus, I have no infomation on which of these antibodies works best in rabbit tissue. If you had unsuccessful experiences with immunogens similar to ab6175 and ab1376, I would suggest to try ab72428.

      We have 3 other nNOS antibodies that are tested in IHC-P and predicted to react with rabbit (based on sequence homology) but have not yet been tested in this species:
      ab79051, ab106417, ab95436

      As for ab3511: This antibody was not tested in IHC-P, thus we do not know how well it would work in this application.

      If you decide to try any of the 4 untestedantibodies, you can use our testing discount program. For UNTESTED species and/or applications, we have established a testing discount program. Here is a brief description of how it works:

      The testing discount program is for customers who like to use an antibody/protein/kit on an untested species/application. You would purchase the antibody at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount for ONE antibody/protein at the full price (100%) of the antibody/protein you have tested, or a full price discount for the amount paid for the kit. The terms and conditions applicable to this offer can be found here: https://www.abcam.com/collaborationdiscount.
      This programapplies to ELISA kits and other kits we offer.

      I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

      Use our products? Submit an Abreview. Earn rewards!
      https://www.abcam.com/abreviews

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      Question

      BATCH NUMBER AGC 0 ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM Only background staining, without specific staining of neurons. SAMPLE Rat brain PRIMARY ANTIBODY Abcam ab6157 (sheep anti rat), dileted in cold fish gelatine 5%. Incubation overnight at room temperature. We tried the following dilutions: 1:200, 1:500, 1:1000, both on PFA2% and PFA 4% perfused brains' sections. After the primary antibodies, the sections were washed 4 times in PBS (+tween and TritonX) DETECTION METHOD Fluorescence POSITIVE AND NEGATIVE CONTROLS USED Negative - omission of primary. Positive should not be needed as the rat brain should be a strong positive control ANTIBODY STORAGE CONDITIONS Aliquotes were made as soon as the antiboy was delivered, and then stores at -80 C FIXATION OF SAMPLE PFA 2% and 4% perfused rat brains, no postfixation. Cut on cryotome (10 micrometer thickness) ANTIGEN RETRIEVAL none. PERMEABILIZATION STEP Triton X in PBS BLOCKING CONDITIONS Cold fish gelatine 5% 30 minutes SECONDARY ANTIBODY Invitrogen Donkey anti sheep Alexa 633 1:200 with DAPI 4 mcl/ml, incubation time 1 h, then washing as for the primary antibody HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 2 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes

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      Answer

      I'm sorry to hear you are experiencing problems with ab6175, I have received feedback from the laboratory that informed me that your protocol is very similar to ours and we would expect it to work, except the addition of tritonx100 in the antibody dilution buffer which significantly helps penetration of the antibody into the tissue. Indeed rat brain is a very good positive control and in our hands the antibody works at 1:2000 (with some background occasionally). This suggests that the antibody has been damaged during shipping or storage. If you purchased the antibody in the last 90 days I can arrange for a replacement vial or refund/credit note if you can provide me with your order details,

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      Question

      Regarding Ab6175, would you expect this antibody to cross react with rabbit eNOS?

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      Answer

      All the information available for this product is listed on the on-line datasheet (price, datasheet, publication, suitability, cross-reactivity).

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      Question

      What is a good paraffin-embedded tissue to use as positive-control? Thank you in advance. Carlo A. Beltrami, M.D.

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      Answer

      A positive control tissue for this antibody is the thoracic spinal cord (of rat). Pre-ganglionic sympathetic neurons stain for NOS.

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      Question

      Reading the questions I realized that on the 22 Nov 2000 was asked whether this Ab detects: 1. INDUCIBLE NOS (= iNOS not bNOS=nNOS) 2. in paraffin embedded slices. Only the second part of the question was answered.

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      Answer

      We cannot provide a definitive answer to this query. However, it is unlikely that our antibody to bNOS recognizes inducible NOS.

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      Question

      I require an antibody against NOS in rabbits - what would you recommend?

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      Answer

      This antibody, ab6175, is known to react with bNOS in rabbits.

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      Question

      does it work in mouse?

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      Answer

      Yes the antibody does cross react with mouse.

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      Question

      Is this antibody is useful to detect expression of inducible nitric oxide synthase in paraffin embedded sections of human brain? Thank you for your help.

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      Answer

      This anti-NOS antibody has not, to our knowledge, been tested in paraffin sections, but we would be surprised if this antibody did not work in paraffin. It is an antibody that is well suited to immunohistochemistry and we have found it works well for human brain (frozen sections).

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