Overview

  • Product name
    Anti-ADAM12 antibody - Aminoterminal end
    See all ADAM12 primary antibodies
  • Description
    Rabbit polyclonal to ADAM12 - Aminoterminal end
  • Host species
    Rabbit
  • Specificity
    This antibody recognizes latent unprocessed and processed forms of ADAM12, but does not react with other ADAMs.
  • Tested applications
    Suitable for: Flow Cyt, ICC/IF, WB, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Pig
  • Immunogen

    Synthetic peptide corresponding to Human ADAM12 (N terminal).

  • Positive control
    • Cell lysate from human fetal fibroblasts (treated with TNF alpha).

Properties

Applications

Our Abpromise guarantee covers the use of ab39155 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
Flow Cyt Use a concentration of 2 µg/ml. PubMed: 19213876

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

 

ICC/IF Use at an assay dependent concentration.
WB 1/1000 - 1/5000. Predicted molecular weight: 100 kDa. A recommended starting concentration for Western blots is 1/1000 when using colorimetric substrates such as BCIP/NBT and 1/5000 for chemiluminescent substrates. EDTA/EGTA treatment of tissues or lysates is required to see latent zymogen.
IHC-Fr Use at an assay dependent concentration. PubMed: 23671573

Target

  • Function
    Involved in skeletal muscle regeneration, specifically at the onset of cell fusion. Also involved in macrophage-derived giant cells (MGC) and osteoclast formation from mononuclear precursors.
  • Tissue specificity
    Isoform 1 is expressed in placenta and skeletal, cardiac, and smooth muscle. Isoform 2 seems to be expressed only in placenta or in embryo and fetus. Both forms were expressed in some tumor cells lines. Not detected in brain, lung, liver, kidney or pancreas.
  • Sequence similarities
    Contains 1 disintegrin domain.
    Contains 1 EGF-like domain.
    Contains 1 peptidase M12B domain.
  • Domain
    The cysteine-rich domain supports cell adhesion through syndecans and triggers signaling events that lead to beta-1 integrin-dependent cell spreading. In carcinomas cells the binding of this domain to syndecans does not allow the integrin-mediated cell spreading.
    The conserved cysteine present in the cysteine-switch motif binds the catalytic zinc ion, thus inhibiting the enzyme. The dissociation of the cysteine from the zinc ion upon the activation-peptide release activates the enzyme.
  • Post-translational
    modifications
    The precursor is cleaved by a furin endopeptidase.
  • Cellular localization
    Secreted and Cell membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • A disintegrin and metalloproteinase domain 12 antibody
    • ADA12_HUMAN antibody
    • ADAM 12 antibody
    • ADAM metallopeptidase domain 12 antibody
    • ADAM12 antibody
    • CAR10 antibody
    • Disintegrin and metalloproteinase domain-containing protein 12 antibody
    • MCMP antibody
    • MCMPMltna antibody
    • Meltrin alpha antibody
    • Meltrin-alpha antibody
    • Metalloprotease disintegrin 12 transmembrane antibody
    • MLTN antibody
    • MLTNA antibody
    • OTTHUMP00000046766 antibody
    see all

Images

  • Anti-ADAM12 antibody - Aminoterminal end (ab39155) at 1/1000 dilution + Human heart tissue lysate - total protein (ab29431) at 10 µg

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/3000 dilution

    Predicted band size: 100 kDa
    Observed band size: 84 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 117 kDa, 42 kDa, 50 kDa. We are unsure as to the identity of these extra bands.

References

This product has been referenced in:
See all 7 Publications for this product

Customer reviews and Q&As

1-10 of 12 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (adipose stem cells)
Permeabilization
Yes - Triton 0.25%
Specification
adipose stem cells
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Mar 07 2017

Question
Answer

On our datasheet for ab39155 we suggest using human fetal fibroblasts as a positive control. Otherwise other positive controls which can be used are: Hela cells, A431 cells or human chondrosarcoma.

Read More

Answer

Vielen Dank für Ihre Email und für Ihr Interesse an unseren Produkten.
Unserem Wissen nach wurde ab39155 bisher noch nicht in IHC-Fr getestet und auch nicht in dieser Anwendung publiziert.
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 ab39155 abdecken, und gegen eine erneute Bestellung eines weiteren Produkts von uns verrechnet werden.
Um von diesem Angebot profitieren zu können, folgen Sie bitte diesen Schritten:
1.) Bestätigen Sie mir bitte, dass Sie ab39155 in IHC-Fr 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 IHC-Fr.
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 ein anderes Produkt zu dem gleichen Preis wie ab39155 bei uns bestellen (halten Sie bei der Bestellung bitte den Rabattcode 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.

Read More

Question
Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number "1131954" ab56366.

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.

I wish you the best of luck with your research.

Read More

Answer

No problem, I will send the product ab56366.

Could you let me know the purchase order number of ab39155?

Thanks!

Read More

Answer

Thank you for your email.

Lab has confirmed that the immunogen for ab115203 and ab28747 is indeed corresponds to N-terminal end and is in signal peptide region, so these antibodies might not be suitable.

Regarding ab56366 this antibody is tested only with recombinant protein used as immunogen; this hasn't yet been tested with endogenous lysates or cell. However If you are interested in using this antibody, then I can offer you Abreview discount along with this replacement. Please click the following link for more information about 100% Abreview discount.

https://www.abcam.com/index.html?pageconfig=resource&rid=11998&viapagetrap=collaborationdiscount

Thirdly, I can offer you a vial of ab39155 from different lot. It may well be that the problem you are experiencing due to damaged vial etc.

Could you please let me know how you would like to proceed?

Read More

Question

Product code: 39155
Lot number: gr3280-1
Inquiry: I've been using this antibody to validate a mass spec experiment whereby ADAM12 was identified as a potential target. We have performed RT-PCR to confirm its expression. I brought this antibody to use for validation primarily by flow cytometry as the data sheet supports the use of this antibody by flow and it also recognises an N-terminal extracellular epitope within the protein. However i have failed to get this antibody to work by flow cytometry despite numerous attempts on multiple human cell lines (all of which are positive by mass spec and RT-PCR). I have tried using trypsin and cell disociation buffer (preserve cell surface epitopes) for cell release from flasks and have used either no blocking (PBS alone) or FCS or BSA as a blocking agent. I have always run the flow using other positive markers that require the same rabbit secondary antibody with positive results. I have tried using down to a 1:50 dilution of the antibody (suggested is 1:500) with no change in result. As the data sheet suggests it can also be used by western blotting i have also tried using the antibody for this purpose twice using a concentration of 1:1000 or 1:500 and 5% non-fat milk for blocking. Again i have achieved no positive staining, although probing for other targets alongside ADAM12 with an identical rabbit-HRP secondary have been successful. Reading the data sheet i was a bit concerned by the size of the product quoted for the shown western blot as the predicted band size is 100kDa, but the shown blot has a major size band of 42 kDa and minor bands at 50 and 84kda. Please could you advise what the next steps are or if you have an alternative antibody that might be more appropiate?

Read More
Answer

Thank you for taking time to complete our questionnaire and for contacting us. I am sorry to hear this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.

Having reviewed the protocol details, I believe this product should have given satisfactory results. It appears that you may have received a faulty vial.

I apologize for the inconvenience and am pleased to offer you a free of charge replacement, credit note, or refund in compensation.

Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

Read More

Question

1) Abcam product code ab39155 2) Abcam order reference number or product batch number 3) Description of the problem detected bands differfrom the expected molcula weight of the molecule 4) Sample preparation: Type of sample (whole cell lysates, fraction, recombinant protein…):whole ell lysate Lysis buffer RIPA buffer Protease inhibitors: yes Phosphatase inhibitors yes Reducing agent yes Boiling for ≥5 min? yes 5 min Protein loaded ug/lane or cells/lane 50ug/lane Positive control yes heart cell lysate Negative control no 5) Percentage of gel 12% Type of membrane nc Protein transfer verified yes Blocking agent and concentration TBS/tween0.1%+milk0.05% Blocking time 1min Blocking temperature room temperature 6) Primary antibody (If more than one was used, describe in “additional notes”) :ab39155 Concentration or dilution 1:5000 and 1: 10000 Diluent buffer TBS/tween0.1%+milk0.05% Incubation time 10min (snap millipore) Incubation temperature: room temperature 7) Secondary antibody: anti-rabbit IgG (sc2054) Species:goat Reacts against: rabbit Concentration or dilution 1:2000 Diluent buffer TBS/tween0.1%+milk0.05% Incubation time 10min (snap millipore) Incubation temperature: room temperature Fluorochrome or enzyme conjugate:hrp 8) Washing after primary and secondary antibodies:yes Buffer TBS/tween0.1% Number of washes 3 9)Detection method chemiluminescence 10) How many times have you run this staining?1 Do you obtain the same results every time? What steps have you altered to try and optimize the use of this antibody?

Read More
Answer

Thank you for taking the time to provide us with the further details. I apologize for the delay of my reply. I am very sorry, that you are experiencing problems with two of our antibodies. I do actually suspect that you have no signal of Adam12 and the bands at 50kD are unspecific. Indeed, I am not aware of any isoform of adam12 around that molecular weight. On Swissprot, there are several isoformes described, all however above 50kD. http://www.uniprot.org/uniprot/O43184 I am therefore sorry not being able to help you to îdentify this band. I can recommend however to add protease inhibitors to the samples in order to exclude that these bands might be degradation products. I have looked throught the details provided, which will also enable us to investigate this case and will provide us with vital information for monitoring product quality. Having reviewed this case, I would like to offer some suggestions to help optimize the results from these two antibodies. I would also appreciate if you can confirm some further details. In the event that we can not solve the problems with these suggestions (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund. 1.) I have noticed that the blocking has been performed in a 0.05% Milk solution. I suppose this has been done for one hour. I can strongly recommend to test also another blocking solution, such as 5% BSA. Also, I do normally recommend to use also a 5% milk solution. Indeed, changing the blocking solution can drastically influence and improve the results. Please see the image on the datasheet of ab9385 as an example. Click here (or use the following: https://www.abcam.com/index.html?datasheet=9385). 2.) I can also suggest to use the primary antibodies in concentration of 1/1000. 3.) As mentinned above, I can recommend to include protease inhibitors in the samples if this has not yet been done sone. If the ab29431 has been used, can you please confirm that the lysat has been stored correctly? Thank you! 4,) I would also apprecaite if you can confirm the quality of the secondary antibody: Does the secondary antibody alone not give such a similar background? Does the secondary antibody work well with other primary antibodies? I hope this information is helpful, and I thank you for your cooperation. I am looking forward to hear back from you with your comments and/or results.    

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (NIH3T3)
Specification
NIH3T3
Fixative
Paraformaldehyde
Permeabilization
Yes - Triton-X-100
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Dec 17 2008

1-10 of 12 Abreviews or Q&A

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

Sign up