Overview

  • Product name

  • Description

    Rabbit polyclonal to DLK-1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, WBmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat, Cow
  • Immunogen

    Synthetic peptide corresponding to Human DLK-1 aa 350 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab23442)

  • Positive control

    • This antibody gave a positive signal in E18.5 Mouse Muscle Wild Type Tissue Lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab21682 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 1/150. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.
WB 1/2000. Detects a band of approximately 45,48 kDa (predicted molecular weight: 41 kDa).

Abcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product.

Target

  • Function

    May have a role in neuroendocrine differentiation.
  • Tissue specificity

    Found within the stromal cells in close contact to the vascular structure of placental villi, yolk sac, fetal liver, adrenal cortex and pancreas and in the beta cells of the islets of Langerhans in the adult pancreas. Found also in some forms of neuroendocrine lung tumor tissue.
  • Sequence similarities

    Contains 6 EGF-like domains.
  • Post-translational
    modifications

    N- and O-glycosylated.
  • Cellular localization

    Membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • Adipocyte differentiation inhibitor protein antibody
    • Brevideltin antibody
    • Delta like 1 homolog antibody
    • Delta like homolog antibody
    • Delta like protein antibody
    • Delta1 antibody
    • DLK 1 antibody
    • DLK antibody
    • DLK-1 antibody
    • DLK1 antibody
    • DLK1_HUMAN antibody
    • FA 1 antibody
    • FA1 antibody
    • Fetal antigen 1 antibody
    • pG2 antibody
    • Preadipocyte factor 1 antibody
    • Pref 1 antibody
    • Pref antibody
    • Pref1 antibody
    • Protein delta homolog 1 antibody
    • Secredeltin antibody
    • ZOG antibody
    see all

Images

  • Image courtesy of Human Protein Atlas

    ab21682 staining DLK-1 in human bone marrow within the haemopoietic tissue and not the adipocytes. Paraffin embedded human bone marrow was incubated with ab21682 (1/150 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.

    ab21682 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org.

  • All lanes : Anti-DLK-1 antibody (ab21682) at 1/2000 dilution ((5% Milk blocking))

    Lane 1 : E18.5 Mouse Muscle Wild Type Tissue Lysate
    Lane 2 : E18.5 Mouse Muscle DLK1 Knockout Tissue Lysate

    Lysates/proteins at 5 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 41 kDa
    Observed band size: 45,48 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 15 kDa (possible cleavage fragment), 55 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 3 minutes


    This blot was produced using an 8% Tris-glycine gel under the Tris-glycine buffer system. The gel was run at 120V for 1 hour 15 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Milk before being incubated with ab21682 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

     

    Abcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.

  • Immohistochemical analysis of formaldehyde-fixed paraffin-embedded human placental tissue sections, labelling DLK-1 with ab21682 at a dilution of 1/1000 incubated for 20 minutes at 25°C. Blocking was with hydrogen peroxide at 3% incubated for 10 minutes at 25°C. The secondary used was a gekko ant-rabbit polyclonal HRP conjugate used undiluted.

    See Abreview

References

This product has been referenced in:

  • Yang W  et al. Adiponectin promotes preadipocyte differentiation via the PPAR? pathway. Mol Med Rep 17:428-435 (2018). Read more (PubMed: 29115433) »
  • Suffee N  et al. Atrial natriuretic peptide regulates adipose tissue accumulation in adult atria. Proc Natl Acad Sci U S A 114:E771-E780 (2017). Read more (PubMed: 28096344) »
See all 15 Publications for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Bone)
Permeabilization
No
Specification
Bone
Blocking step
Vetor Blocker as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 23°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Sep 19 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (placenta)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate, pH6, 100C, 20 min
Permeabilization
No
Specification
placenta
Blocking step
H2O2 as blocking agent for 10 minute(s) · Concentration: 3% · Temperature: 25°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Aug 19 2015

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (neurons)
Permeabilization
Yes - 0.2% Tween 20
Specification
neurons
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 24°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Apr 23 2014

Answer

Muchas gracias por tu paciencia.

Lamento la espera. Mis compañeros del laboratorio me han confirmado que este anticuerpo solo lo han testado en células MES en inmunocitoquímica, y en WB en lisados de MEL-1, F9 y Mouse EG.

Incluso en nuestra imagen en la datasheet parece haber cierta tinción filamentosa. Estamos investigando el caso, y haremos las pruebas adicionales pertinentes.

Tenemos otros dos anticuerpos anti DLK monoclonales, testados en ICC en células humanas:

https://www.abcam.com/DLK-antibody-3A10-ab119930.html

https://www.abcam.com/DLK-antibody-MM0514-9D8-ab89908.html]

Si quisieras recibir alguno de ellos en compensación por las molestias de forma gratuita, no dudes en comunicármelo. Alternativamente, y dado que ab21682 está cubierto por la garantía Abpromise, si lo prefieres podemos directamente reembolsarte el importe del mismo, mandarte un anticuerpo contra otra proteína, o emitir un vale sin caducidad para futuras compras con nosotros.

De nuevo te pido disculpas, y quedo a la espera de tu respuesta. ¡Que pases un buen día!

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Question
Answer

x
Je suis ravie que vous souhaitiez participer a notre offre et tester ab21682 en cytométrie de flux. Le code de réduction correspondant est xx . Ce code de réduction est utilisable sous 120 jours et pour l’achat d’un autre anticorps primaire.
Vous devez dans un premier temps soumettre une Abreview avec vos résultats en cytométrie de flux en notant ce code dans la section « Additional notes ». Ce code sera ainsi activé et vous pourrez par la suite contacter notre service clientèle par téléphone pour passer votre prochaine commande, en vous munissant des informations suivantes : code de réduction, numéro de commande de ab21682.
Nous restons à votre disposition pour plus d’informations concernant cette offre promotionnelle. Dans l’attente de recevoir votre Abreview nous vous souhaitons bonne chance dans vos recherches.
Termes et Conditions : www.abcam.com/collaborationdiscount.

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Question

First issue: thank you for the offer: we would indeed appreciate a lot to receive a new vial of ab110363 and a purification kit for tissue culture supernatant ab109207 to replace the lost material. This could help us a lot.
Second issue: Questionnaire for ab21682:
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 ab21682, rabbit anti-DLK1

2) Abcam order reference number or product batch number : GR31847-1
3) Description of the problem: high aspecific background, impossible/difficult to pick up the DLK1 signal. Conclusion: antibody is not specific, but gives a very particular intracellular/perinuclear staining pattern (=golgi/ER?)
4) Sample preparation:
Species: rat pancreatic beta cells
Type of sample: Fresh frozen sections, perfusion fixed frozen sections, PFA/formalin fixed paraffin embedded sections, cells in culture, other:
Sample preparation : fixation in formol
Positive control: unstimulated cells (MIX1), negative for DLK1
Negative control : cells treated with specific compound to trigger DLK1 expression (MIX2), confirmed at level of qPCR (80.000 fold increase at RNA level), ICC with 5 other rabbit anti DLK1 antibodies, including ab110636
5) Fixation step
Yes/No
If yes: Fixative agent and concentration : formaldehyde 3.7% in PBS
Fixation time: 30 minutes to 1 day
Fixation temperature: cold room 4°C
6) Antigen retrieval method: none, not required for the other tested antibodies, which all were very specific (see picture)
7) Permeabilization method:
Did you do a permeabilization step (details please) or add permeabilizing agent in any dilution buffers?
Permeabilizing agent and concentration:
TritonX100, 0.5% in PBS
8) Blocking agent (eg BSA, serum…): BSA 3%
Concentration
Blocking time: 30 to 60 minutes
Blocking temperature room temperature
9) Endogenous peroxidases blocked? No, not relevant
Endogenous biotins blocked? No, not relevant
10) Primary antibody (If more than one was used, describe in “additional notes”) :
Concentration or dilution 1/250, 1/1000
Diluent buffer: PBS
Incubation time: overnight, 4°C
11) Secondary antibody:
Species: donkey anti rabbit
Reacts against:
Concentration or dilution: 1/3000
Diluent buffer PBS
Incubation time: 30 minutes
Fluorochrome or enzyme conjugate: 488
12) Washing after primary and secondary antibodies:
Buffer: PBS+3% BSA
Number of washes : 3x 5 minutes
13) Detection method: 488 fluorescence microscope – DAPI for nuclei
14) How many times have you run this staining? 3x, in parallel with other DLK1 antibodies (ab110636, santacruz and an non commercial ab)
Do you obtain the same results every time? yes
What steps have you altered to try and optimize the use of this antibody? Not, other antibodies have been used as possitve control, we have no need to optimize this ab further.
Document attachment: see enclosed picture

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Answer

Thank you for these precisions.
We are still working on your complaint for ab21682 (anti-DLK1) and we will come back to you as soon as possible.
As requested, I have issued a free of charge replacement with the order number *******.
To check the status of the order please contact our Customer Service team and reference this number.
Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

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Question

Dear,
We have a number of concerns:
1) We were recently adviced to buy the antibody purification kit (ab102784) to remove azide from the monoclonal ab110636 (see letter below), which we need to be able to use in cell culture as indicated. In the mean time we have bought this kit together with a FITC-labelingskit and two new batches of DLK1 antibodies (ab110636 and ab21682). After clean up with ab102784, it was found that ab110636 did not bind to the adviced collums, infact the antibody eluted already with the first step, the bidingbuffer. Only a minor fraction could be recovered in the washing buffers and elution buffers. An overview of our concentration measurements is provided in the attached file. This finding was also confirmed by ICC wish failed to provide a signal on a positive cell sample.
For ab21682 whitch was purified in parallel we did not notice this problem, but we only need this antibody as a negative control. It is Ab110636 which we should obtain in a purified, azide free form. What is possible?
One of the goals was also to label the purified ab110636 with de labeling kit, which now cannot be used.
2) ab21682 has been tested for ICC using 2 conditions, one negative and a second with a stimulated expression of DLK1, it was compared for its specificity to 5 other DLK1 antibodies (see attached figure). Our conclusion is that ab21682 is not specific for its target, but shows a high aspecific background staining, which is problematic since this antibody is mentioned in a few publications, and is as such the first choice to buy ffrom the website. It is however doubtfull whether this antibody can be used for ICC, and should not be sold for this purpose, since the "background" signal could be misinterpreted. I did not test it on paraffin samples for IHC
Ab110636 on the contrary is highly specific.
What could you propose?

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Answer

Thank you very much for your email and your patience.
In regards to the first issue, with the purification of ab110363, I had contacted the laboratory. There is nothing special with this clone. The only thing which might could explain the result is, that the ab110636 you received is at a low concentration. Therefore it could be too dilute to bind and elute efficiently. I can therefore propose to send you a new vial of ab110363 and a purification kit for tissue culture supernatant ab109207 for free, so you can perform the experiment. Please let me know whether you would like to accept this offer.
In regards to the ab21682 staining in ICC, I would be pleased to investigate this further. Can you please provide me with an image and the protocol details? I have attached a questionnaire which will help you to put the protocol details together. Once I have received this information, it will be easier for me to investigate and troubleshoot the results, as well as initiate internal actions if necessary. The specificity of the product is obviously very important. If the product is faulty, we will also replace it for you.
Thank you very much for your cooperation. I am looking forward to hear back from you.

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Answer

Thank you for calling Abcam last night.

I have spoken to my colleagues about your wish to return the 2 vials of ab21682. Since they were purchased more than 2 weeks ago, we cannot accept them as a return, I am sorry about that. I understand that these antibodies are expensive, so below is a discount that will give you 30% off the next antibody that you purchase:

Discount Code : *******
Expiration Date: ***********

If there is anything else I can help you with, please let me know.

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Answer

Thank you for contacting us regarding this possibility. I have contacted the lab regarding the possibility that this product is recognizing the extracellular domain. I’m afraid we have no testing data to show the specificity for the intracellular domain as we do not specifically test for this. We only have the fact that the immunogen is raised against the intracellular region and the peptide blocking on the WB. It could be possible that it may be cross reacting with the extracellular domain but it is just not something we currently test for. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information. Use our products? Submit an Abreview. Earn rewards! https://www.abcam.com/abreviews

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Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Embryonic stem cell-derived hepatocyte precursors)
Permeabilization
No
Specification
Embryonic stem cell-derived hepatocyte precursors
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Fixative
Paraformaldehyde

Dr. Irina Drobinskaya

Verified customer

Submitted Feb 12 2009

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