Overview

  • Product name
    Anti-Neurexin II alpha antibody
  • Description
    Rabbit polyclonal to Neurexin II alpha
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-P, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Rat, Human
    Predicted to work with: Mouse
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 250 - 350 of Rat Neurexin II alpha.

    Read Abcam's proprietary immunogen policy (Peptide available as ab34449.)

  • Positive control
    • normal adult human cerebral cortex SKNSH cell line

Properties

Applications

Our Abpromise guarantee covers the use of ab34245 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-P 1/700. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
WB 1/400.
ICC/IF Use a concentration of 5 µg/ml.

Target

  • Relevance
    Neurexin 2 alpha belongs to the Neurexin family, a group of neuronal proteins that function as cell adhesion molecules during synaptogenesis and intracellular signaling. Mammalian genomes contain just three neurexin genes. However, many (>200) neurexin isoforms may be expressed through the use of two alternative promoters and alternative splicing at up to five different positions.
  • Cellular localization
    Membrane; single-pass type I membrane protein.
  • Database links
  • Alternative names
    • Neurexin 2 antibody
    • Neurexin II antibody
    • NRXN2 antibody

Images

  • (Image courtesy of Human Protein Atlas)
    ab34245 staining Neurexin II alpha protein in normal adult human cerebral cortex. Brown color indicates presence of protein, blue color shows cell nuclei. Paraffin embedded human cerebral cortex tissue was incubated with ab34245 (1/700 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.

    .
  • Anti-Neurexin II alpha antibody (ab34245) at 1/400 dilution + whole cell lysate prepared from murine N2A cells at 20 µg

    Secondary
    Goat polyclonal conjugated to Dylight 800 at 1/20000 dilution

    Performed under reducing conditions.

    Predicted band size: 185 kDa
    Observed band size: 160 kDa
    why is the actual band size different from the predicted?



    Samples were blocked using 0.2% casein for 1 hour at 37°C.

    See Abreview

  • ICC/IF image of ab34245 stained SKNSH cells. The cells were 4% formaldehdye 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 (ab34245, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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.

References

This product has been referenced in:
  • Harkin LF  et al. Neurexins 1-3 Each Have a Distinct Pattern of Expression in the Early Developing Human Cerebral Cortex. Cereb Cortex 27:216-232 (2017). Read more (PubMed: 28013231) »
  • Brito-Moreira J  et al. Interaction of amyloid-ß (Aß) oligomers with neurexin 2a and neuroligin 1 mediates synapse damage and memory loss in mice. J Biol Chem 292:7327-7337 (2017). Read more (PubMed: 28283575) »
See all 4 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Question

1) Abcam product code ab34245
2) Abcam order reference number or product batch number : GR10984-1
3) Description of the problem: Detection of a band with lower MW than predicted in datasheet (˜50 kDa)
4) Sample preparation:
Type of sample (whole cell lysates, fraction, recombinant protein…): rat hippocampal primary culture
Lysis buffer : RIPA
Protease inhibitors: protease inhibitor cocktail from Pierce
Phosphatase inhibitors : none
Reducing agent : beta-mercapto
Boiling for ≥5 min? yes/no: no
Protein loaded ug/lane or cells/lane: 20 ug
Positive control : none
Negative control : none

5) Percentage of gel
Type of membrane : PVDF
Protein transfer verified: Ponceau
Blocking agent and concentration: Odyssey blocking buffer (Lycor)
Blocking time : 1 hour
Blocking temperature : RT (˜25 oC)

6) Primary antibody (If more than one was used, describe in “additional notes”) : ab34245
Concentration or dilution : 1:400
Diluent buffer : Odyssey blocking buffer (Lycor)
Incubation time: overnight
Incubation temperature: 4 oC

7) Secondary antibody: Odyssey
Species:
Reacts against: rabbit
Concentration or dilution : 1:15,000
Diluent buffer : Odyssey blocking buffer (Lycor)
Incubation time: 2 hours
Incubation temperature: RT
Fluorochrome or enzyme conjugate: 800 (green, Odyssey)

8) Washing after primary and secondary antibodies:
Buffer: TBS + Tween 0.1%
Number of washes: 4 times, 5 min each
9) Detection method: Odyssey (Lycor)
10) How many times have you run this staining? 2 times
Do you obtain the same results every time? yes
What steps have you altered to try and optimize the use of this antibody? none

Read More
Answer

Thank you for your enquiry regarding ab34245 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody.

The expected band size for mature Neurexin II alpha should be around 185 kDa. The band observed at 50 kDa either represent some degradation product or different splice variants. Since RIPA buffer has been used for sample preparation, as long as the proteinase inhibitors are freshly prepared before the experiments, the degradation can be ruled out.

It is worth mentioning that cells in primary culture tend to dedifferentiate very quickly and the protein expression profile may be different from cells isolated from tissues or from well established cell cultures.

As our datasheet indicates, normal adult human cerebral cortex SKNSH cell line and murine N2A cells were used to test this antibody. It would be advisable to run a good positive control alongside with the samples to compare the expression pattern of Neurexin II alpha.

I would also suggest checking the literature what type isoforms are reported to be expressed in rat hippocampal cells. According to Swiss-Prot database, 18 (!) different isoforms produced by alternative promoter usage and alternative splicing. Please visit this site for further information:

http://www.uniprot.org/uniprot/Q63374

If you need any further assistance in the future, please do not hesitate to contact me.

Read More

Answer

Thank you for your enquiry regarding ab34245.

According to Swiss-Prot database, there seems to be some splice variants and isoforms Neurexin II alpha. You may wish to take a look at this site for further information:

http://www.uniprot.org/uniprot/Q9P2S2

Could you please provide some further details of the protocol used and complete the following form (attached as a word document). It would be much appreciated if you could attach an image to the response.

Thank you for your understanding and co-operation in this matter. I look forward to hearing from you soon and resolving this issue as soon as possible.

Read More

Answer

Thank you very much for your interest in ab34245. To our knowledge, ab34245 has not been tested in Rat (it is only predicted). Therefore, I can offer a discount off a future purchase if you buy ab34245 now, test it in Rat and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free PRIMARY ANTIBODY. 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 test ab34245 in Rat. I will then send a discount code. This code must be issued before purchasing ab34245 so please wait for my reply before ordering. 2. Purchase ab34245 either by phone, fax, or online (www.abcam.com). 3. Test it in Rat. 4. Let us know 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 any PRIMARY ANTIBODY ordered and the discount code is valid for 4 months after issue. We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab34245 turns out to be unsuitable for Rat, you will still receive the discount on your next purchase after your Abreview has been submitted. 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
Application
Western blot
Sample
Mouse Cell lysate - whole cell (N2A cells)
Loading amount
20 µg
Specification
N2A cells
Gel Running Conditions
Reduced Denaturing
Blocking step
casein as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.2% · Temperature: 37°C

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Submitted Dec 23 2010

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