Overview

  • Product name
    Anti-Syntaxin antibody [STX01 (HPC-1)]
    See all Syntaxin primary antibodies
  • Description
    Mouse monoclonal [STX01 (HPC-1)] to Syntaxin
  • Host species
    Mouse
  • Specificity
    In hippocampus, the antibody recognizes the stratum radiatum, the layer formed by the fibers of denate granule cells, as they synapse onto hippocampal pyramidal cells. In rat retina, it stains the membrane of the amacrine cell bodies and inner plexiform layer. It recognizes some neurons early in development and can be detected on migrating amacrine cells in embryonic retina.
  • Tested applications
    Suitable for: WB, IP, ICC/IFmore details
  • Species reactivity
    Reacts with: Rat, Human
  • Immunogen

    Recombinant fragment corresponding to Rat Syntaxin aa 3-225.

  • Positive control
    • IMR-5 cells Cerebral cortex tissue sections. SH-SY5Y cell lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab3265 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
WB 1/100. Detects a band of approximately 31 kDa (predicted molecular weight: 35 kDa).
IP 1/200.
ICC/IF 1/50.

Target

Images

  • All lanes : Anti-Syntaxin antibody [STX01 (HPC-1)] (ab3265) at 1 µg/ml

    Lane 1 : Human brain tissue lysate - total protein (ab29466)
    Lane 2 : Human spinal cord tissue lysate - total protein (ab29188)
    Lane 3 : Spinal Cord (Rat) Tissue Lysate
    Lane 4 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 35 kDa
    Observed band size: 35 kDa
    Additional bands at: 48 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 1 minute
  • ICC/IF image of ab3265 stained PC12 cells. The cells were 4% formaldehyde 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 (ab3265, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse 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.

References

This product has been referenced in:
  • Toft-Bertelsen TL  et al. Regulation of Ca2+ channels by SNAP-25 via recruitment of syntaxin-1 from plasma membrane clusters. Mol Biol Cell 27:3329-3341 (2016). WB ; Mouse . Read more (PubMed: 27605709) »
  • Shen C  et al. The trans-SNARE-regulating function of Munc18-1 is essential to synaptic exocytosis. Nat Commun 6:8852 (2015). Read more (PubMed: 26572858) »
See all 8 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Answer

Thank you for contacting us. I am sorry to hear that you are having problems with the 3 antibodies ab53723, ab11104 and ab3265. Unfortunately, ab53723 has not been tested for IHC-Frozen, horse and pig samples. Also, ab11104 has not yet been tested in IHC-Frozen and horse samples. It is the same situation for ab3265 which has not yet been tested for IHC in general as well as horse and pig samples. (We refer to fresh frozen tissue as IHC, and to cultured cells as ICC/IF). Thus, we do not know if and how well these antibodies would perform for the application/species used. The correct seconday antibodies would have been an anti-mouse secondary for both ab3265 and ab11104, and an anti-rabbit secondary for ab53723. What fluorophore have you been using? What exactly was the problem that you encountered for each antibody? And what antibody dilution, blocking and detection step did you use? Please let me know. Also, it might be advisable to use a fixation step to ensure the proteins are kept in the correct location. Was a permeabilization step included? The crossreactivity of an antibody depends mostly on the sequence homology between the immunogen and the protein sequence of interest. Do you have the horse and pig sequences for all 3 proteins? That way I could check which antibody might be most suitable for horse and pig samples. I checked the antibodies we currently have in the catalog: For Syntaxin, we have 4 other antibodies that have been tested with pig samples: ab113986 (FITC), ab112200 (biotin), ab112198 and ab24731. But none of these have been tested in IHC or with frozen sections or with horse samples. For Synaptobrevin, we have 3 other antibodies that have been tested with pig samples: ab24735, ab18015 and ab18013. Again, none of these have been tested with IHC or horse samples. For SNAP25, we have ab24732 that has been tested with pig samples, ab117083 that has been predicted to react with pig samples. None of these however were tested with horse or IHC-Fr samples. We have also 4 antibodies that were tested in frzen sections (IHC-Fr), but not yet in pig or horse: ab68859, ab72396, ab68858 and ab72394. 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 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 at the full price (100%) of the antibody you have tested. The terms and conditions applicable to this offer can be found here: https://www.abcam.com/collaborationdiscount. As for secondary antibodies, the choice depends on the host of the primary antibody (e.g. for a rabbit-raised primary antibody, the secondary shuld be an anti-rabbit) as well as what conjugation you prefer to use or which detection system is available to you (e.g. which wavelength can be detected). I look forward to hear back from you with more details so that I can assist you further.

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Answer

Thank you very much for your patience. Product ab3265 is raised against Syntaxin 1 of rat origin. Since there are only 2 amino acids that fall within an extracellular region, the chances of Syntaxin 1 (HPC-1) mapping to this region are quite low (very low). However, because we do not characterize the exact eptiope of this antibody, we cannot guarantee the exact region the antibody maps to. I have updated the datasheet. I apologize for the inconvenience. I hope this information will be helpful. Should you have any other question please do not hesitate to contact me.

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Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Horse Tissue sections (cranial cervical ganglion)
Specification
cranial cervical ganglion
Fixative
Formalin
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: High pH Vector labs H-3301
Permeabilization
No

Abcam user community

Verified customer

Submitted Feb 09 2011

Answer

Thank you for getting back to me. The details of the epitope region (extracellular) are provided by the source of this antibody and is shown on the datasheet. Please see the link below. I hope this information helps. Please do not hesitate to contact me should you require further assistance.

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Question
Answer

Thank you for your enquiry. I am sorry but the epitope sequence ab3265 remains proprietary. I can however tell you that the epitope resides in the extracellular domain of the protein. I hope this information helps. Please do not hesitate to contact me should you require further assistance.

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Answer

Thank you for your enquiry. All the information we have on species cross reactivity is specified on the datasheet. To our knowledge, cross reactivity with mouse has not yet been tested. Should you decide to go ahead and purchase this product, please let us know how you get on and in return we will forward a reward of your choice, typically an Amazon gift voucher.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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