Overview

  • Product name
  • Description
    Rabbit polyclonal to NGF
  • Host species
    Rabbit
  • Specificity
    Less than 1% cross-reactivity against recombinant human Brain Derived Neurotrophic Factor, Neurotrophin 3 and Neurotrophin 4/5 by ELISA.
  • Tested applications
    Suitable for: ICC/IF, Dot blot, Neutralising, WB, IHC-FoFr, IHC-Fr, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Human
    Does not react with: Cow
  • Immunogen

    Native mouse salivary gland NGF purified by modification of the method of Mobley et al (1976). Antigen purity was greater than 95% by PAGE.

  • Positive control
  • General notes
    This antibody has been shown to be useful for a variety of techniques and its specificity has been demonstrated by immunoblot.

Properties

Applications

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.

Application Abreviews Notes
ICC/IF Use a concentration of 10 µg/ml.
Dot blot Use at an assay dependent concentration.
Neutralising Use a concentration of 2 - 10 µg/ml. 2ug/ml of ab6199 can neutralize 100ng/ml mouse NGF.
WB Use a concentration of 1 µg/ml.
IHC-FoFr 1/300 - 1/5000. (see Abreview on perfusion fixed tissue for detailed protocol).
IHC-Fr Use at an assay dependent concentration.
IHC-P 1/500.

Target

  • Function
    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).
  • Involvement in disease
    Neuropathy, hereditary sensory and autonomic, 5
  • Sequence similarities
    Belongs to the NGF-beta family.
  • Cellular localization
    Secreted.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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

Images

  • ab6199 staining perfusion fixed rat brain and dorsal root ganglion by IHC-Fr. Animals were pre-perfused with Tris buffer pH 10, followed by 4% paraformadehyde and 15% of a saturated solution of picric acid. The brains were post-fixed in the same fixative overnight, cryoprotected in 20% sucrose for 24 hours, frozen and cut with a cryostat. Free floating immunostaining was performed. An Alexa Fluor ® 488 conjugated goat anti-rat antibody was used as the secondary.

    The image shows the staining obtained with this antibody using direct fluorescence in the rat cortex and dorsal root ganglion. The staining is not only of the cell body of the cortical neurons but a part of their processes.

    See Abreview

  • ab6199 staining NGF in Human tendon tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with Dako FLEX Peroxidase blocking for 5 minutes at room temperature; antigen retrieval was by heat mediation in Dako high pH. Samples were incubated with primary antibody (1/250) for 30 minutes. An undiluted HRP-conjugated Goat polyclonal was used as the secondary antibody.

    See Abreview

  • 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.

  • ab6199 at a 1/500 dilution staining rat brain tissue sections by Immunohistochemistry (Formalin-fixed paraffin-embedded sections). The tissue section was paraformaldehyde fixed and blocked with 2% BSA prior to incubation with the antibody for 24 hours. Bound antibody was detected using a biotinylated goat anti-rabbit IgG antibody.

    This image is courtesy of an Abreview submitted by Grazyna Niewiadomska.

    See Abreview

References

This product has been referenced in:
  • Fu F  et al. NDUFA4 enhances neuron growth by triggering growth factors and inhibiting neuron apoptosis through Bcl-2 and cytochrome C mediated signaling pathway. Am J Transl Res 10:164-174 (2018). Read more (PubMed: 29423002) »
  • Jasim H  et al. Saliva as a medium to detect and measure biomarkers related to pain. Sci Rep 8:3220 (2018). Read more (PubMed: 29459715) »
See all 26 Publications for this product

Customer reviews and Q&As

1-10 of 28 Abreviews or Q&A

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

Mr. JUNGHO YOON

Verified customer

Submitted Aug 16 2017

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

Abcam user community

Verified customer

Submitted Aug 15 2014

Question
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)

Read More

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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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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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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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.

Read More

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.

Read More

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
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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