Anti-NGF 抗体 (ab6199)

Reviews (8)Q&A (20)References (47)

Key features and details

  • Rabbit polyclonal to NGF
  • Suitable for: ICC/IF, WB
  • Reacts with: Mouse, Rat, Chicken, Human
  • Isotype: IgG

製品の概要

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab6199 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
ICC/IF Use a concentration of 10 µg/ml.
WB Use a concentration of 1 µg/ml.

ターゲット情報

  • 機能

    Nerve growth factor is important for the development and maintenance of the sympathetic and sensory nervous systems. Extracellular ligand for the NTRK1 and NGFR receptors, activates cellular signaling cascades through those receptor tyrosine kinase to regulate neuronal proliferation, differentiation and survival. Inhibits metalloproteinase dependent proteolysis of platelet glycoprotein VI (PubMed:20164177).

  • 関連疾患

    Neuropathy, hereditary sensory and autonomic, 5

  • 配列類似性

    Belongs to the NGF-beta family.

  • 細胞内局在

    Secreted.
  • Information by UniProt

  • 参照データベース

    see all

  • 別名

    • Beta nerve growth factor antibody
    • Beta NGF antibody
    • Beta-nerve growth factor antibody
    • Beta-NGF antibody
    • HSAN5 antibody
    • MGC161426 antibody
    • MGC161428 antibody
    • Nerve growth factor (beta polypeptide) antibody
    • Nerve growth factor antibody
    • Nerve growth factor beta antibody
    • Nerve growth factor beta polypeptide antibody
    • Nerve growth factor beta subunit antibody
    • NGF antibody
    • NGF_HUMAN antibody
    • NGFB antibody
    • NID67 antibody
    see all

画像

  • Western blot - Anti-NGF antibody (ab6199)
    Western blot - Anti-NGF antibody (ab6199)
    All lanes : Anti-NGF antibody (ab6199) at 5 µg/ml

    Lane 1 : rhNGF protein at 100 µg
    Lane 2 : Mouse salivary gland tissue lysate at 50 µg


    ab6199 detects a strong band at 13 kDa consistent with the expected molecular weight of mature NGF monomer.

  • Western blot - Anti-NGF antibody (ab6199)
    Western blot - Anti-NGF antibody (ab6199)
    Anti-NGF antibody (ab6199) at 1 µg/ml + Active human NGF full length protein (ab69759) at 0.1 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Exposure time: 30 seconds
  • Immunocytochemistry/ Immunofluorescence - Anti-NGF antibody (ab6199)
    Immunocytochemistry/ Immunofluorescence - Anti-NGF antibody (ab6199)
    ICC/IF image of ab6199 stained MEF1 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab6199, 10µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

参考文献 (47)

ab6199 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。

ab6199 は 47 報の論文で使用されています。

  • Zheng Y  et al. Altered expression of glycoprotein non-metastatic melanoma protein B in the distal sciatic nerve following injury. Int J Mol Med 45:1909-1917 (2020). PubMed: 32236569
  • Zhang J  et al. Upregulation of TRPV1 in spinal dorsal root ganglion by activating NGF-TrkA pathway contributes to pelvic organ cross-sensitisation in rats with experimental autoimmune prostatitis. Andrologia 51:e13302 (2019). PubMed: 31074030
  • Nagura N  et al. Nerve growth factor continuously elevates in a rat rotator cuff tear model. J Shoulder Elbow Surg 28:143-148 (2019). PubMed: 30146165
  • Rudnitskaya EA  et al. Alterations of hippocampal neurogenesis during development of Alzheimer's disease-like pathology in OXYS rats. Exp Gerontol 115:32-45 (2019). PubMed: 30415068
  • Zhao Y  et al. L-NBP, a multiple growth factor activator, attenuates ischemic neuronal impairments possibly through promoting neuritogenesis. Neurochem Int 124:94-105 (2019). PubMed: 30629983
View all Publications for this product

レビューと Q&A

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1-10 of 28 Abreviews or Q&A

Western blot abreview for Anti-NGF antibody

 Excellent
Abreviews
Application
Western blot
Sample
Human Cell lysate - whole cell (gastric cancer cell)
Gel Running Conditions
Reduced Denaturing (15% gel)
Loading amount
30 µg
Specification
gastric cancer cell
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1µg/mL · Temperature: 24°C
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Mr. JUNGHO YOON

Verified customer

投稿 Aug 16 2017

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-NGF antibody

 Excellent
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Dako FLEX Peroxidase blocking as blocking agent for 5 minute(s) · Concentration: 100% · Temperature: RT°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Dako High pH
Sample
Human Tissue sections (Tendon)
Specification
Tendon
Permeabilization
No
Fixative
Formaldehyde
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Abcam user community

Verified customer

投稿 Aug 15 2014

Question

Can you suggest a positive control for Western blotting?

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Answer

Several different lysates could be used as a positive control. We have used ab6199 with MEF1 cell line in Western blotting. Alternatively it has also been used with rat brain tissue and should also work with mouse or human brain tissue. We have a range of different brain lysates. These can be viewed under the following catalogue numbers:

Brain (Human) Tissue Lysate - adult normal tissue (ab29466)

Brain (Mouse) Whole Cell Lysate - normal tissue (ab30151)

Brain (Rat) Whole Cell Lysate - normal tissue (ab29475)

Brain (Mouse) Tissue Lysate - normal tissue, 0 days old (ab7188)

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Question


Inquiry: For neutralization studies using this anti-NGF antibody, what control rabbit IgG product do you recommend?

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Answer

Thank you for contacting Abcam.

I beleive the rabbit IgG control below would work very well for your application.

https://www.abcam.com/Rabbit-IgG-ChIP-Grade-ab37415.html



Also, please see the attachment to find out details of our current promotion, of buy one antibody, get a RabMab for free.

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Question

Sehr geehrte Damen und Herren,
gerade haben wir den AB NGF ( ab 6199) erhalten.
In den tech notes wird geraten, glycerol hinzuzugeben.
" add glyceerol to a final volume of 50 %".
Wie soll dies denn verstanden werden. Unser Ab hat ein Volumen von 500 µl.
Könnten sie die Logik dahinter kurz erläutern. Wir haben gerade darüber
schwer gegrübelt
Es gab verschiedene Ansätze und eine Wette dazu. ;-)
Besten Dank für ihre Antwort
Mit freundlichen Grüßen
--

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Answer

Vielen Dank für Ihre Anfrage.
Bitte geben Sie bei einem Volumen von 500ul Antikörper 500ul Glycerin (Glycerol) dazu.
Das ergibt eine Endkonzentration von 50% Glycerin in 1000ul.
Ich hoffe, der Wetteinsatz ist interessant :-)

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Question

Bonjour,
Merci beaucoup pour votre retour. Je suis disponible toute la matinée au
Cordialement,

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Answer

Suite à notre conversation téléphonique, je vous envoie la référence de la publication de Emoto où différentes méthodes de démasquage d'antigène ont été testées et comparées : Emoto et al., J Histochem Cytochem. 2005. 53 (11) : 1311-21.

J'espère que ces informations vous seront utiles. N'hésitez pas à nous contacter de nouveau si vous avez d'autres questions.

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Question

Hi,
I was looking to use the Anti-NGF antibody (ab6199)for Western blotting (reducing conditions)in mouse heart lysates. Thespecification sheet has a picture of a Western blot witha band corresponding to ˜25-37kDa.
Just wondering if you knew a definitive MW that we should be expecting bands at?
Doesthe antibodydetect NGF precursor and mature forms?
Any additionalinformationon the running conditions for the blot shown in the specification sheetwould be greatly appreciated.

Cheers,

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Answer

Thank you for your inquiry.

The calculated molecular weight of NGF is 27kDa according to UniProt (http://www.uniprot.org/uniprot/P01138).

This molecular weight will be higher when the protein is phosphorylated and glycosylated. According to the SwissProt page there are multiple modifications in NGF.

We have customer feedback in form of an Abreview that will provide some more protocol information for you:

https://www.abcam.com/index.html?datasheet=6199&tab=abreviews&intabreviewid=29302

There are also publications that might be helpful:

Zhao LH et al. Cornel iridoid glycoside improves memory ability and promotes neuronal survival in fimbria-fornix transected rats. Eur J Pharmacol 647:68-74 (2010). WB; Rat. PubMed: 20826142
Maezawa I et al. Rett syndrome astrocytes are abnormal and spread MeCP2 deficiency through gap junctions. J Neurosci 29:5051-61 (2009). WB; Mouse. PubMed: 19386901



I hope this information is helpful and wish you good luck with your experiments.

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Question


I am a current user of theAnti-NGF (ab6199), I was wondering whether I could obtain some further information regarding this product.



1. What is the optimal antigen retreival method.



2.The pH of the antigen retreival solution, if any.



3.The optimal dilutionof the Anti-NGF (ab6199).



4. References on usage of the Anti-NGF (ab6199).

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Answer

Thank you for your enquiry.

Here are answers to your questions:

1.So far we know heat mediated antigen retrieval has worked.

2. citrate buffer pH 6,0

3. Optimal dilution must be determined emperically, but 5ug/ml has been used succesfully.

4. Here is a link to published papers using ab6199:
https://www.abcam.com/NGF-antibody-ab6199-references.html

I hope this is helpful. Please contact us again if you have any further questions.

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Question

Product code: 6199
Lot number: GR74133-1

Inquiry: Hello I'm trying to make the '5 % acetic acid buffer containing 0.9 % NaCL (ASB) at pH 3' for perfusion prior to fixing as your protocol states. Could you let me know a recipie for this please? Does the acetic acid buffer contain NaAcetate as well? I have tried to look at the reference you provide (Zhou et al 1994) but can not access it. Kind regards

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Answer

Thank you for your inquiry about a reference that was mentioned by a customer that submitted an Abreview (http://www.ncbi.nlm.nih.gov/pubmed/7815824)


Unfortunately, we do not have access to the specific article that was mentioned, nor do we have subscriptions to these journals. If you would like additional information regarding specific protocols or samples used, we would like to suggest contacting the authors of the article directly or ask your library to order the article for you.

For advice regarding our products and protocols, we are happy to assist you. Please do not hesitate to contact us if you would like additional information.

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Question

Désolée j'ai oublié de vous préciser le tissus, c'est du cerveau humain.

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Answer

Merci pour votre réponse et ces précisions.

J'ai de nouveau regardé vos marquages et je validerais personnellement ces résultats. Les deux anticorps montrent un bon marquage spécifique et lescontroles négatifs avec l'isotype sont d'aucune ambiguité.

J'espère que ceci vous sera utile. Merci de nous avoir contactés. N'hésitez pas à nous contacter de nouveau si vous avez d'autres questions.

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1-10 of 28 Abreviews or Q&A

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