Overview

  • Product name
    Anti-Doublecortin antibody
    See all Doublecortin primary antibodies
  • Description
    Goat polyclonal to Doublecortin
  • Host species
    Goat
  • Specificity
    ab113435 is expected to recognize all four reported isoforms (NP_000546.2; NP_835365.1; NP_835364.1; NP_001182482.1). Reported variants represent identical protein: NP_835364.1 and NP_835366.1.
  • Tested applications
    Suitable for: WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Horse, Cat, Dog, Pig
  • Immunogen

    Synthetic peptide:

    C-KTSANMKAPQS

    , corresponding to internal sequence amino acids 151-161 of Human Doublecortin (NP_835365.1; NP_835364.1; NP_001182482.1.), or amino acids 232-242 of Human Doublecortin (NP_000546.2).

  • Positive control
    • Mouse fetal Brain lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab113435 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 Use a concentration of 0.01 - 0.03 µg/ml. Detects a band of approximately 45 kDa (predicted molecular weight: 49 kDa).
IHC-P Use a concentration of 2 - 4 µg/ml.

Target

  • Function
    Seems to be required for initial steps of neuronal dispersion and cortex lamination during cerebral cortex development. May act by competing with the putative neuronal protein kinase DCAMKL1 in binding to a target protein. May in that way participate in a signaling pathway that is crucial for neuronal interaction before and during migration, possibly as part of a calcium ion-dependent signal transduction pathway. May be part with LIS-1 of an overlapping, but distinct, signaling pathways that promote neuronal migration.
  • Tissue specificity
    Highly expressed in neuronal cells of fetal brain (in the majority of cells of the cortical plate, intermediate zone and ventricular zone), but not expressed in other fetal tissues. In the adult, highly expressed in the brain frontal lobe, but very low expression in other regions of brain, and not detected in heart, placenta, lung, liver, skeletal muscles, kidney and pancreas.
  • Involvement in disease
    Defects in DCX are the cause of lissencephaly X-linked type 1 (LISX1) [MIM:300067]; also called X-LIS or LIS. LISX1 is a classic lissencephaly characterized by mental retardation and seizures that are more severe in male patients. Affected boys show an abnormally thick cortex with absent or severely reduced gyri. Clinical manifestations include feeding problems, abnormal muscular tone, seizures and severe to profound psychomotor retardation. Female patients display a less severe phenotype referred to as 'doublecortex'.
    Defects in DCX are the cause of subcortical band heterotopia X-linked (SBHX) [MIM:300067]; also known as double cortex or subcortical laminar heterotopia (SCLH). SBHX is a mild brain malformation of the lissencephaly spectrum. It is characterized by bilateral and symmetric plates or bands of gray matter found in the central white matter between the cortex and cerebral ventricles, cerebral convolutions usually appearing normal.
    Note=A chromosomal aberration involving DCX is found in lissencephaly. Translocation t(X;2)(q22.3;p25.1).
  • Sequence similarities
    Contains 2 doublecortin domains.
  • Cellular localization
    Cytoplasm.
  • Information by UniProt
  • Database links
  • Alternative names
    • DBCN antibody
    • Dbct antibody
    • DC antibody
    • DCX antibody
    • DCX_HUMAN antibody
    • Doublecortex antibody
    • Doublin antibody
    • FLJ51296 antibody
    • Lis X antibody
    • Lis-X antibody
    • Lissencephalin X antibody
    • Lissencephalin-X antibody
    • Lissencephaly X linked antibody
    • Lissencephaly X linked doublecortin antibody
    • LISX antibody
    • Neuronal migration protein doublecortin antibody
    • OTTHUMP00000023859 antibody
    • OTTHUMP00000023860 antibody
    • OTTHUMP00000216315 antibody
    • OTTHUMP00000216316 antibody
    • SCLH antibody
    • XLIS antibody
    see all

Images

  • Anti-Doublecortin antibody (ab113435) at 0.01 µg/ml + Mouse Fetal Brain lysate (in RIPA buffer) at 35 µg

    Developed using the ECL technique.

    Predicted band size: 49 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cerebellum labeling Doublecortin with ab113435 at 2µg/ml. Microwaved antigen retrieval with citrate buffer pH 6, HRP-staining.

References

This product has been referenced in:
  • Tu M  et al. Notch1 Signaling Activation Contributes to Adult Hippocampal Neurogenesis Following Traumatic Brain Injury. Med Sci Monit 23:5480-5487 (2017). IHC-P ; Rat . Read more (PubMed: 29150595) »
  • Jing YH  et al. Adult neural stem cell dysfunction in the subventricular zone of the lateral ventricle leads to diabetic olfactory defects. Neural Regen Res 12:1111-1118 (2017). IHC . Read more (PubMed: 28852393) »
See all 2 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Brain sections / Dentate Gyrus of the Hippocampus)
Antigen retrieval step
None
Permeabilization
Yes - PBST 3 x 5mn RT
Specification
Brain sections / Dentate Gyrus of the Hippocampus
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Aug 01 2017

Answer

Vielen Dank für Ihre Antwort. Ich habe Ihnen einen Anti-Doublecortin Antikoerper aus der Ziege herausgesucht. Click here (or use the following: https://www.abcam.com/index.html?datasheet=113435). Doublecortin färbt keine Glia -Zellen an (siehe Paper im Anhang). Allerdings wird es am stärksten im Neonatalen Gehirn expremiert. Wenn die Zellen aus einem alteren Tier stammen, kann das Signal dementsprechend schwächer sein. Unseres Wissens nach wurde der ab113435 bisher noch nicht in ICC getestet. Falls Sie dies selbst testen möchten, kann ich Ihnen zur Zeit ein spezielles Angebot über einen 100%igen Abreview-Rabatt anbieten. Bei diesem Angebot bekommen Sie einen Rabatt für eine zukünftige Bestellung, wenn Sie uns ein Abreview mit dem Testresultat zusenden. Der Rabatt würde den ganzen Wert von ab113435 abdecken, und gegen eine erneute Bestellung eines weiteren primären Antikörpers von uns verrechnet werden. Um von diesem Angebot profitieren zu können, folgen Sie bitte diesen Schritten: 1.) Bestätigen Sie mir bitte, dass Sie ab113435 in ICC testen möchten. 2.) Bitte bestellen Sie erst nach Erhalt des Rabattcodes. 3.) Bestellen Sie den Antikörper wie üblich per Telefon, Email oder Fax 4.) Testen Sie den Antikörper in ICC. 5.) Teilen Sie uns das Ergebnis Ihres Tests durch ein Abreview mit und notieren Sie den Discount Code in dem Feld "additional notes" Unter der folgenden URL können Sie mehr über unser Abreview System erfahren: https://www.abcam.com/abreviews 6.) Der Rabattcode ist nach dem Abschicken des Abreviews aktiv, und Sie können einen anderen primären Antikörper zu dem gleichen Preis wie ab113435bei uns bestellen (halten Sie bei der Bestellung bitte den Rabattcode und die Bestellnummer von ab113435 bereit). Bitte beachten Sie, dass der Rabattcode innerhalb von 4 Monaten nach Ausstellung eingelöst werden muss. Der Rabattcode wird gültig unabhängig davon, ob Ihr Ergebnis positiv oder negativ ist. Die Bedingungen zu unserem 100% Abreview Rabatt können Sie unter dem folgenden Link nachlesen: https://www.abcam.com/collaborationdiscount Ich hoffe, diese Informationen helfen Ihnen weiter. Falls Sie einen Rabattcode erhalten möchten oder weitere Fragen haben, zögern Sie bitte nicht, sich wieder an mich zu wenden.

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Question

Hi,

Here are the answers to your IHC questionnaire about the double-cortin antibody. Any advice would be much appreciated.

Thanks
xxxxx

IHC Questionnaire
It will take approximately 5 minutes to complete the questionnaire.
Please fill-in all fields so that we can better assist you
1) Abcam product code
ab113435
2) Abcam order reference number or product batch number
lot: GR74374-2, order ref: 30234634
3) Description of the problem
Antibody used for immunohistochemistry, but only non-specific staining seen.
4) Sample preparation:
Species Mouse
Type of sample: perfusion fixed frozen sections, fixed with 4% PFA, strored in cryoprotectant
Sample preparation:
Positive control
Negative control No antibody
5) Fixation step
Yes
If yes: Fixative agent and concentration 4% paraformaldehyde
Fixation time: 4 hours
Fixation temperature room temperature
6) Antigen retrieval method
7) Permeabilization method:
Did you do a permeabilization step (details please) or add permeabilizing agent in any dilution buffers?
Permeabilizing agent and concentration: triton-X, 0.3% in PBS in all buffers
8) Blocking agent (eg BSA, serum…):
Concentration 10% normal horse serum
Blocking time 1 hour prior to antibody addition, and overnight with antibody
Blocking temperature room temp, then 4oC overnight
9) Endogenous peroxidases blocked? Yes (3% hydrogen peroxide, 10% methanol in distilled water)
Endogenous biotins blocked? No
10) Primary antibody (If more than one was used, describe in “additional notes”) :
Concentration or dilution 1:500, 1:1000 and 1:2000
Diluent buffer PBS with 0.3% triton-X and 10% horse serum
Incubation time 4oC overnight
11) Secondary antibody:
Species: horse
Reacts against: goat
Concentration or dilution 1:200
Diluent buffer PBS with 0.3% triton-X and 10% horse serum
Incubation time 2 hours at room temp
Fluorochrome or enzyme conjugate biotinylated
12) Washing after primary and secondary antibodies:
Buffer PBS
Number of washes 3
13) Detection method DAB and fluorescence
14) How many times have you run this staining? 6
Do you obtain the same results every time? yes
What steps have you altered to try and optimize the use of this antibody? I have altered the antibody concentrations, blocking and buffer solutions.

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Answer

Thank you for getting back to me with those details.
As I have mentioned previously, we have not performed any IHC experiments with this antibody, we would therefore not be sure of how well it would perform in this application.
You mentioned on the phone that one of your colleagues had used a Doublecortin antibody for IHC. Were you able to confirm that it was ab113435?
Having reviewed the protocol you have been using there are a few suggestions I can make which may help to improve the results obtained.
1. You have used PFA to fix the tissue. This may mean that some form of antigen retrieval is required which can be very tricky with brain tissue. In order to avoid this you could fix the tissue in a different fixative such as acetone, methanol or ethanol. Otherwise it may be worth having a look at the following reference for antigen retrieval of frozen sections:
Yamashita S, Okada Y. Application of heat induced antigen retrieval to aldehyde fixed fresh frozen sections. J. Histochem Cytochem. 2005 Nov;53(11):1421-32.
2. I would suggest performing a "no primary" control alongside with the normal staining to see if the secondary antibody is contributing to the non-specific binding. It may also be worthwhile to reduce the concentration of secondary antibody.
3. I would reduce the concentration of hydrogen peroxide used to block the endogenous peroxidases to 0.3 % and it may be worthwhile to perform this block following the incubation of the primary antibody to reduce any potential damage to the epitope.
I hope these suggestions help to understand where the problems may be coming from. Have you tried using the antibody in Western blotting (an application which the anitbody has been used with before). What were the results of this?
We have several other antibodies which may provide better results than the ones you have observed so far, such as rabbit polyclonals ab18723 and ab77450. With both of these antibodies, if you were to have any problems in achieving good staining, you would be guaranteed by the Abpromise, more information of which can be found from the following link:
www.abcam.com/abpromise
I hope this information has been of help. I look forward to recieving your reply.

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Answer

Thank you for contacting us and reporting the difficulty you have found in using the anti-Doublecortin antibody (ab113435) in IHC.
As discussed over the phone, we have not ourselves tested this antibody in IHC and have had no reports of our customers having tried it either. It may therefore be that the antibody is not suitable for this application. However, I would be happy to try to advise you as to how the protocol may be optimised in order to obtain the best results. For this reason, I have attached the questionnaire we discussed over the phone to this email.
If you have managed to find out if the antibody your colleagues were using to stain Doublecortin was ab113435 that would be very interesting to know. And if you are able to share with me the lot number they used I can have a look into this as well.
I look forward to receiving your reply.

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Answer

Thank you very much for your interest in ab113435.

We do not supply samples of our antibodies and so therefore I am unable to send you an aliquot to try out and you are correct that ab18723 is a rabbit polyclonal antibody.

As ab113435 has not been tested in IHC-P. Therefore, I can offer a discount off a future purchase if you buy ab113435 now, test it in IHC-P 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 ab113435 in IHC-P. I will then send a discount code. This code must be issued before purchasing ab113435 so please wait for my reply before ordering.

2. Purchase ab113435 either by phone, fax, or online (www.abcam.com).

3. Test it in IHC-P.

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 ab113435 turns out to be unsuitable for IHC-P 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.

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Answer

Thank you for contacting Abcam.

We have not yet tested ab113435 in IHC on brain sections, therefore we do not have a protocol that has been worked out for this antibody, I have however attached the Abcam general IHC protocol. Also as it has not been tested in this application, I am happy to offer you a testing discount for this antibody, you can find more information on this by following the link below:

https://www.abcam.com/index.htmlpageconfig=resource&rid=11998&viapagetrap=collaborationdiscount



I would recommend that you also look at ab18725, as this antibody has been shown to work in various IHC methods and in multiply species.

Please let me know if you would like to take advantage of our testing discount program or if there is anything else I can help you with.

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