Anti-VEGFC 抗体 (ab9546)

Hello! This antibody should be able to detect both the prescursor and mature VEGFC. Please let me know if I can be of further assistance.

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The rabbit anti-mouse VEGF-A antibody ab51745 did not cross react with human VEGF-C in Western blotting (reduced, non-reduced). According to our results the rabbit anti-rat VEGF-C ab9546 cross reacts with human VEGF-C but not with human VEGF165.

Regarding ab105219 I am sorry, we have not tested this antibody for reactivity with VEGFC.


I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Thank you for contacting us.

I am happy that you have decided to use our antibody for your research purposes. 1 vial of ab9546 will contain 100 ul at 1mg/ml. This will be sufficient for you to perform 10 western blots and 10 immunohistochemistry experiments if you use the amounts indicated below:

Western blot
Stock solution: 100 µg at 1mg/ml
Recommended concentration for western blot: 1 ug/ml

If you incubate your western blot membrane in 5 ml of primary antibody solution, you will need 5 ug of ab9546 per sample. Therefore, for 10 samples you would need 5ug x 10 = 50ug. Since each vial has 100 ug of antibody, you would be left with 50 ug of antibody left for your immunohistochemistry studies.

Immunohistochemistry
Stock solution: 100 µg at 1mg/ml
Recommended concentration for IHC: 10 ug/ml

You need a 1:100 dilution to get a concentration of 10ug/ml. If you use 100 ul of primary antibody solution per sample, you would need 1 ul of antibody per sample. Therefore for 10 samples you would need 1ug x 10 = 10 ug.

In total, this means that for 10 samples of WB and 10 samples of IHC, you would use up a total of 60 ul of antibody from the 100 ul supplied.

Unfortunately, we do not offer discounts for one vial of product. We can arrange discounts on orders for five vials or more. This can be five vials of the same product, or mixed products. If you would like to place a bulk order, please contact our customer services team for further information. However, at the moment we are running an exciting promotion where for any primary antibody bought, you get a rabbit monoclonal antibody for free. Just add the following code to your next order to take advantage of this offer: RABMAB-XBSMG

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

Thank you for contacting us.

Firstly I would like to confirm that you are referring to ab9546 and not ab9456?

ab9546 is a rabbit polyclonal and therefore does not have a specific epitope. This antibody contains a combination of immunoglobulin molecules each identifying a different epito[e. I can however confirm that the antibody is produced in the same way as the antibody used in the publication.

For storage conditions, make sure you read the datasheet carefully and follow the instructions. The antibody should be shipped at 4°C but upon delivery, you should aliquot it and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles

Aliquotting minimizes damage due to freezing and thawing, as well as contamination introduced by pipetting from a single vial multiple times. Aliquots are to be frozen and thawed once, with any remainder kept at 4oC. Upon receiving the antibody, centrifuge at 10,000 x g for 20 seconds to pull down solution that is trapped in the threads of the vial, and transfer aliquots into low-protein-binding microcentrifuge tubes. The size of the aliquots will depend on how much one typically uses in an experiment. Aliquots should be no smaller than 10 µl; the smaller the aliquot, the more the stock concentration is affected by evaporation and adsorption of the antibody onto the surface of the storage vial.

In general if an antibody is stored properly as stated on the datsheet and is kept at -20°C, it will keep for 5 years. If stored at -80°C, it will keep for 10 years. However, please note that these are only guidelines and we can not guarentee how long the products will last. Please only note that our guarentee is only valid for 6 months after the delivery date of the product.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Thank you for contacting us.

VEGF-C analysis has not yet been done on human skin with antibody ab9546. However, this antibody is guarenteed to work in humans and in WB and IHC therefore it should be fine to use ab9456 on
innumohistochemistry and western blotting on human skin samples.

If you would like to test 100 samples in western blot and 100 samples in IHC, you will need the following amounts of this antibody:

Western blot
Stock solution: 100 µg at 1mg/ml
Recommended concentration for western blot: 1 ug/ml

If you incubate your western blot membrane in 5 ml of primary antibody solution, you will need 5 ug of ab9546 per sample. Since each vial has 100 ug of antibody, you could do 20 samples with one vial. Therefore in total, 5 vials of antibody will be needed to do 100 western blot.

Immunohistochemistry
Stock solution: 100 µg at 1mg/ml
Recommended concentration for IHC: 10 ug/ml

If you use 100 ul of primary antibody solution per sample, you can do 100 tests with one vial of antibody.

Therefore you would need 6 vials of ab9456 in total each at £234.00. However, on orders of more than 4 items, it is possible for us to give a discount. If you our interested in this offer please conatact our customer services team by emailing mailto:orders@abcam.com

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

I am very sorry to hear that you have been experiencing problems using this product in the application that you wish.

I would like to reassure you that our Abpromise applies to your complaint since you purchased this product within the guarantee period. This means that in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.

Could you 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.

I can confirm that this antibody has been tested for ELISA and IP in the lab but the IHC-P as tested application has been added to the datasheet based on several specific publications. You may wish to take a look at these papers for further information:

Yan A et al. Mechanisms of lymphatic regeneration after tissue transfer. PLoS One 6:e17201 (2011). IHC-P; Mouse. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=21359148&dopt=Abstract

Wada H et al. Distinct characteristics of circulating vascular endothelial growth factor-a and C levels in human subjects. PLoS One 6:e29351 (2011). IHC-FoFr; Mouse. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=22206010&dopt=Abstract

Machnik A et al. Mononuclear phagocyte system depletion blocks interstitial tonicity-responsive enhancer binding protein/vascular endothelial growth factor C expression and induces salt-sensitive hypertension in rats. Hypertension 55:755-61 (2010). WB, IHC-P; Human. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=20142563&dopt=Abstract

Machnik A et al. Macrophages regulate salt-dependent volume and blood pressure by a vascular endothelial growth factor-C-dependent buffering mechanism. Nat Med 15:545-52 (2009). IHC-P; Rat.

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.

Thank you for your reply.

I was wandering if you had tried using a higher concentration of the antibody? If the antibody binds to the mouse protein with a lower affinity (this is a theory of mine, we have not actually tested it), then you may need to use more antibody.

I’ll keep searching to see if I can determine what the issue. It may be that for whatever reason the vial of antibody that you have may not be able to detect that low levels of protein. In which case I would be happy to replace or refund the cost of the antibody.

Let me know if you have, or if using more antibody proves to be successful.

Thank you for your patience while I was looking into this matter.

I have been searching our notes and I cannot find the reference number for the exact VEGF-C protein that was used to so the test ELISA, but I suspect that it was a protein that encoded recombinant rat VEGF-C, http://www.uniprot.org/uniprot/O35757.

I aligned the mouse and rat sequences and they are 97% homologus, but it may be that the antibody has a lower affinity to the mouse protein compared to the rat and so it may not be able to detect that low a level of mouse protein.

I am going to look into this a little further, but wanted to let you know that we are investigating this matter and I also had one question: what was the concentration of antibody you were using, was it 2.5ug/ml?

I look forward to your reply and helping to resolve this issue.

Thank you for contacting us.

Because we carry over 70,000 products, it isn't feasible for us to keep small sample sizes of our products.
We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase.

A discount can be offered if you purchase 5 products or more.

Please find below the selection of antibodies that, I think, may be suitable for your work, assuming you work on Human tissue samples:

VEGF : https://www.abcam.com/ab46154 (Rabbit polyclonal), https://www.abcam.com/ab1316 (Mouse monoclonal) or https://www.abcam.com/ab68334 (Mouse monoclonal). All very well characterised and tested in IHC-P.
VEGFA : https://www.abcam.com/ab105219 (Mouse monoclonal). Tested in IHC-P.
VEGFB : https://www.abcam.com/ab58461(Rabbit polyclonal). Not yet tested in IHC-P. Eligible for a testing discount (see details below).
VEGFC : https://www.abcam.com/ab9546 (Rabbit polyclonal), tested in IHC-P.
VEGFD : https://www.abcam.com/ab103685 (Rabbit polyclonal), tested in IHC-P




To our knowledge, ab58461 has not been tested in IHC-P. Therefore, I can offer a discount off a future purchase if you buy ab58461 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 ab58461 in IHC-P. I will then send a discount code. This code must be issued before purchasing ab58461 so please wait for my reply before ordering.

2. Purchase ab58461 either by phone, fax, or online (https://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.
Please remember that submission of the Abreview is sufficient for the discount code to become active.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab58461 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: https://www.abcam.com/collaborationdiscount.

Following up my previous e-mail, I received the following from our source for this antibody:

"The proteolytic processing of mammalian VEGF-C is a complicated step and described before. Therefore the work by Joukov et al. (EMBO J 16, pp3898-11, 1997) must be read very carefully. The antibody made against the mature VEGF-C region will certainly recognize the full unprocessed, partially processed and mature VEGF-C. If you take whole mouse tissue you may come up with the detection of the more intracellular unprocessed or partially processed forms and not so much the released forms fully processed by (extracelluar?) proteases. Cell culture supernatants from mouse trachea cells will probably give a total different picture."

Please let me know if you have questions or concerns.