Anti-VEGF antibody (ab46154)

Thanks for your answer,but there is another question.If the protein sample was treated by reducing agent,most of them should be momomer.But we could not see the 22kDa band.Did you identify the band or have other data to support your idea ?

ANSWER:

I agree that if the samples were treated with reducing agent, then unreduced VEGF is a bad explanation for the absence of 22 kDa monomer,

The only other explanation I know of for VEGF appearing at 40 - 45 kDa is the possibility that the samples contain only L-VEGF. This is an alternative transcript with 180+ more amino acids than the "normal" transcript. The long isoforms are listed at this link for the UniProt entry for human VEGF:

http://www.uniprot.org/uniprot/P15692

This article has a discussion of long VEGF but I am not able to access the full article:

Nuclear localization of long-VEGF is associated with hypoxia and tumor angiogenesis.
Biochem Biophys Res Commun. 2005 Jun 24;332(1):271-8.
PMID: 15896327

Here is the abstract: and some small images:
http://www.sciencedirect.com/science/article/pii/S0006291X0500896X

This article also shows a blot with ab46154 showing a band at 43 kDa for tumor tissue in Figure 2:

Xu D et al. Heparan sulfate regulates VEGF165- and VEGF121-mediated vascular hyperpermeability. J Biol Chem 286:737-45 (2011).
http://www.jbc.org/content/286/1/737.long

There are several other references for ab46154 that used the antibody for western blotting listed on the datasheet.

If you have not detected a band at 20 - 22 kDa and are concerned about the specificity of the antibody, I will be happy to discuss your protocol and samples. If you expect a band at 22 kDa for you samples, I will also be happy to discuss a replacement or refund, if you have purchased the antibody within the past 6 months.

Can you please send me some information about your samples and your western blot protocol? If you are testing cell lysates, what are the cells, or tissues? Are the samples reduced and denatured? How much protein do you load in the gel, and how much antibody do you incubate with the blot?

I look forward to your reply.

Technical service,
I want to know whether the band of 43kDa is the band of VEGF protein in WB by using Anti-VEGF antibody (ab46154) .The VEGF protein band should be about 20 KDa ,if the exeriment is performed under reducing condition,because it is in state of monomer.Thanks!

ANSWER:

Thank you for contacting us.

The laboratory that validated this antibody for western blotting suspects that it is detecting the homodimer, not the monomer at 22 kDa. The samples were treated with a reducing agent but the lab thinks the treatment did not sufficiently reduce the dimer. I will find out if there is more recent data, as I expect the blot was repeated. I will forward this data to you when I receive it.

Please do not hesitate to contact us if you need any more advice or information.

Phone call: Has this antibody already been tested on frozen sections?

ANSWER:

DISCOUNT CODE: xx Ablaufdatum: 20. Juli 2012

Es freut mich zu hören, dass Sie unser Angebot unseren Antikörper ab46154 auf gefrorenen Schnitten zu testen, annehmen möchten. Wie am Telefon besprochen, können Sie den Gutscheincode zum Kauf eines weiteren Antikörpers einsetzen, sobald Sie uns ein Abreview für diesen Antikörper eingereicht haben. Der Discountcode hat die Nummer xx, deckt den Wert von ab46154 ab und kann zum Kauf weiterer Produkte eingesetzt werden.

Der Code wird gültig, sobald Sie uns ein Abreview für diesen Antikörper über den Test auf gefrorenen Schnitten (IHC-Fr) eingereicht haben. Bitte geben Sie diesen Code auch im Abschnitt “Additional Notes” des Abreviews mit an, so dass wir wissen, dass sich dieses Abreview auf die Gutscheinaktion bezieht. Der Code wird dadurch aktiviert. Bitte kontaktieren Sie bei der nächsten Bestellung unsere Kundendienst mit dem Code und der ursprünglichen Bestellnummer.

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Hello,
we are just starting a new project looking at VEGF. I am interested in trying the anti-VEGF antibody (ab46154) and was wondering if this will work in sheep? Is it possible for me to get a sample to test?
Thank you,

ANSWER:

Thank you for contacting us with your question.

Ab46154 has not been tested in sheep samples, but I ran an alignment of the sheep protein sequence to the immunogen, and there is 96% identity between the sequences. So, it is quite possible that this antibody will cross-react but we can't say for sure until it's tested.

We only have the full size vials so I don't have a small sample to send out, however we do have a testing discount program that you might be interested in. In this program, you would first purchase ab46154 at full price and test it in sheep, but in exchange for submitting an Abreview with your data, we will give you one free antibody on a future purchase. So if ab46154 does not work for some reason, you can either try a different VEGF antibody for free or you can select an antibody to a different target. If you would like to take part in this program, I would be happy to send more information.

Please let me know if you have any further questions or if there is anything else that I can do for you.

Thank you for your reply.

This customer would like to apply the credit note.
Would you please help provide the credit note?


Thank you very much.
Best regards,

ANSWER:

Thank you for confirming this information and for your help and cooperation with this case.

As requested, I have asked our accounting department to issue you with a credit note. This can then be redeemed against the invoice of a future order.

As usual if you have any further questions regarding this credit note, please contact the accounts department by email at creditcontrol@abcam.com. The reference number, or credit ID, is #####. Please refer to this number in any correspondence with the accounting department.

I would like to wish the customer good luck with their research. The technical team is always at your service, should you require further expert advice.