Good evening guys, I did a comparision between the mAb anti-M13:HRP from GE (the one everybody use and unfortunatelly it is discontinued) with a brand new version from Santa Cruz Biotech (SC-53004-HRP).

Basically it works as good as the one from GE at the same dilution. I also evaluated the signal of the same titration curve using decreasing amounts of the secondary antibody and the performance is very good.

The product sc-53004 HRP is available at 200 ug/mL (1 mL) and It worked very nicely at 1/5K dilution (100 uL/well). Enough reagent for 96-well 500 plates.

Hello Martin,

Besides the anti-M13-HRP from GE I have also good experience with anti-p8(M13)-biotin (http://www.antibodies-online.com/antibody/125965/anti-M13+Bacteriophage+G8P+Capsid+antibody+Biotin/?utm_source=partner&utm_medium=biocompare&utm_campaign=listing&utm_content=165 or the same alternative https://www.acris-antibodies.com/antibodies/primary-antibodies/m13-bacteriophage-g8p-capsid-bm5516b.htm), although you also need a secondary conjugate streptavidin-HRP for detection. In western blots these antibodies seem to work very well and appears to be highly sensitive.

Otherwise, I also usually use anti-M13-HRP from GE and I am very surprised that this conjugate is discontinious...

Good luck,

Peter

Dear Martin & Peter,

Others have posted the same question not too long ago, and clearly the loss of the GE antibody is an issue for many. If you have had good results with the Abs from antibodies-online or Acris or others, we encourage you to not just mention it here, but to also post a review at http://pabmabs.com/wordpress/. Other researchers will thank you!

I have had many issues with this, and I have also not completely solved it yet.

1) Do not buy the Abcam antibody. It is shitty, expensive, and cannot be diluted more than 1/20 (compared to normal dilutions of 1/1000 to 1/5000). This means you pay something like GBP 400 to test HALF an ELISA plate.

2) I have changed to a system using an anti-m13 mouse antibody as primary antibody and then an anti-mouse-HRP secondary antibody (https://www.jacksonimmuno.com/catalog/products/115-005-205). The secondary antibody work WELL, but I am yet to find a good primary anti-m13 mouse antibody

Thank you so much Andreas and Peter for the advice! I will try Andreas' option and probabilly I will make a polyclonal in rabit and conjugate it to HRP in house. Thank you so much!!!

Hi Martin, may I suggest you to look at this discussion https://www.researchgate.net/post/HRP_Anti-M13_Monoclonal_antibody_for_phage_ELISA#view=594b8d3af7b67eccce6e9422. I have the same problem than you and they suggest me 2 alternative 

Good Morning, I asked a sample of this mAb to Santa Cruz Biotech and I compared it side by side with the mAb (without HRP) from GE and worked beautifully. I titrated the antibody and it is has to be used 5X more concentrate but still it was 1/5000 dilution from the stock (200 ug/mL).

The only issue is the extra step with the anti-mouse HRP (1/5000) but I am very happy that this is a good alternative. The company also has the HRPversion of this antibody but they didnt have samples of it, but I am going to buy it and I will post a graph making a comparation with the mAB:HRP from GE.

https://www.scbt.com/scbt/product/m13-major-coat-protein-antibody-rl-ph2

HRP/Anti-M13 Monoclonal antibody for phage ELISA?. Available from: https://www.researchgate.net/post/HRP_Anti-M13_Monoclonal_antibody_for_phage_ELISA#view=594b8d3af7b67eccce6e9422

As I saw in other post Larbi Mokhtari already ordered this HRP version of the antibody, please keeo us updated.

Thanks for the info. Martin. Please consider also posting a review of the mAb on pabmabs.com

Hi Martin, unfortunately after wainting more than a month, Santa Cruz told me that the product didn't pass quality control testing, so I canceled my order and now I am going to to order from Sino Biological (cat: 11973-MM05)

uhhh thats so bad! Let us know how this one works please.

The one that I did  test was the unlabeled and worked very nicelly but you have an extra step with the antimouse:HRP.

Best luck

PS. I also placed an order for the same item from Santa Cruz

I use the anti-Fd from sigma and then an anti-mouse-HRP for detection.

Dear all,

Regarding the anti-m13 (HRP) selection. Based on the discussion above, I already got a quotation for SC-53004-HRP. I saw Martin A Rossotti also order the item 7 months ago, did u have any feedback on this since Larbi Mokhtari cancelled the order due to QC issue. In Malaysia I can't find any distributor for Sino Biological (cat: 11973-MM05T-H).

Thanks & best regards,

Chew

Good evening guys, I did a comparision between the mAb anti-M13:HRP from GE (the one everybody use and unfortunatelly it is discontinued) with a brand new version from Santa Cruz Biotech (SC-53004-HRP).

Basically it works as good as the one from GE at the same dilution. I also evaluated the signal of the same titration curve using decreasing amounts of the secondary antibody and the performance is very good.

The product sc-53004 HRP is available at 200 ug/mL (1 mL) and It worked very nicely at 1/5K dilution (100 uL/well). Enough reagent for 96-well 500 plates.

Hi Martin. Thanks a lot for sharing this valuable information on pAbmAbs.com. Best wishes form the pAbmAbs team

Thanks Martin for feedback regarding SC-53004-HRP.

Very unfortunately, I faced same problem with Larbi Mokhtari, pending for delivery for more than 2 months, distributor also can't promise when the item can release from Santa Cruz. Now the only way to solve it is seeking for Sino Biological (cat: 11973-MM05T-H).

So bad to hear that! Fortunatelly in my case the product arrived very fast to Ottawa, Canada. Best luck!

Thnx a lot, Martin!

You did a really good job, performing a direct comparison between 'gold standard' GE and Santa Cruz antibodies.

I think SC should provide you with lifetime support of SC-53004-HRP!)

Hi everybody.

I ordered the anti-M13 HRP from santa cruz (sc-53004) 2 months ago and I received it well.

But all my tests with it were bad.

I need to dilute the antibody to 1/50 or 1/100 or put a lot of phages in elisa wells (1.10e12 phages/well) to see a signal. AND this antibody has a very high background (see attached).

Has anybody the same experience than mine. Do you know why there is a huge backgroud when my well is coated with PBS, and no background when it's coated whith 1.10e9 phages/well???

Thanks a lot.

Hey Aurelie Schneider it is so bad to hear this, and I am also so dissaponted of the only antibody that looked decent in the market.

Recentelly I had a similar issue, I did run out of the first vial I bought 6 month a, I recieved a new one and I did the titration to verify that everything was ok. comparing the new one side by side with the older alicuot. In the assay I also included the awesome and sadly discontinued unlabelled mAb from GE. As you can see in the pdf file, the signal of the second vial (I guess anotehr batch) was much weaker. I think it could be a problem in the peroxidation that inactivate the mAb (? just guessing but honestelly I dont know).

Before ordering the HRP labelled anti M13, I requested to Santa Cruz a sample of the unlabelled one. So do the same, ask them a sample and share with them whats happening and when you recieve ittry it in the same experiment using a secondary anti mouse.:HRP I know it is annoying because it add an extra step and you can not use it against antigens with mouse Fc, but this seems to be the only solution.

To reduce the background always helps to add tween and another protein in the dilution buffer but I guess the problem is because the hight concnetration that need to be used to get a signal. Another silly question. Best luck

Hi, Progen is producing the B62-FE2 as HRP conjugate: https://www.progen.com/anti-m13-fd-f1-filamentous-phages-mouse-monoclonal-b62-fe2-hrp-conjugate.html

Best wishes,

Michael