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Sheep Anti-Mouse IgG H&L (HRP) (ab102458)

Submit a review Q&A (3)References (1)

Key features and details

  • Sheep Anti-Mouse IgG H&L (HRP)
  • Conjugation: HRP
  • Host species: Sheep
  • Isotype: IgG
  • Suitable for: ICC, ELISA, IHC-P, WB

製品の概要

  • 製品名

    Sheep Anti-Mouse IgG H&L (HRP)
    IgG 二次抗体 製品一覧

  • 由来種

    Sheep

  • ターゲット生物種

    Mouse

  • 特異性

    By immunoelectrophoresis and ELISA this antibody reacts specifically with mouse IgG and with light chains common to other mouse immunoglobulins. No antibody was detected against non-immunoglobulin serum proteins. This antibody may cross react with IgG from other species.

  • アプリケーション

    適用あり: ICC, ELISA, IHC-P, WBmore details

  • 標識

    HRP

製品の特性

  • 製品の状態

    Liquid

  • 保存方法

    Shipped at 4°C. Store at +4°C.

  • バッファー

    pH: 6.8
    Constituents: 0.2% BSA, 0.05% CMIT/MIT based preservative
  • Concentration information loading...

  • 精製度

    Immunogen affinity purified

  • 特記事項（精製）

    This antibody was isolated by affinity chromatography using antigen coupled to agarose beads and conjugated to HRP.

  • ポリ/モノ

    ポリクローナル

  • アイソタイプ

    IgG

  • 研究分野

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab102458の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
ICC
Use at an assay dependent dilution.
ELISA
1/10000 - 1/100000. (Primary)
IHC-P
1/200 - 1/5000.
WB
1/10000 - 1/50000. with a chemoluminescence detection method. If using a colorimetric method, dilute 1/5000 - 1/30000.
特記事項
ICC
Use at an assay dependent dilution.
ELISA
1/10000 - 1/100000. (Primary)
IHC-P
1/200 - 1/5000.
WB
1/10000 - 1/50000. with a chemoluminescence detection method. If using a colorimetric method, dilute 1/5000 - 1/30000.

プロトコール

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

参考文献 (1)

ab102458 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab102458 は 1 報の論文で使用されています。

  • Ramirez-Alvarado M  et al. Assessment of renal response with urinary exosomes in patients with AL amyloidosis: A proof of concept. Am J Hematol 92:536-541 (2017). PubMed: 28295502

レビューと Q&A

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1-3 of 3 Abreviews or Q&A

Question

OK, I have a little more idea of how best to use the sheep anti-mouse IgG-HRP antibody, ab102458. It turns out, from the panel of dot-blots that I did, that the Abcam secondary is just about 2.5 times stronger than the one we usually use, and if I adjust for this, I have better results and less fading.

I could not duplicate the fading I was seeing on my Westerns by using the dot blot, probably because the signal would be much more highly concentrated in a single Western blot band than dispersed in a dot, and I still do get some fading when I probe my Westerns, but I do not think it is the fault of your antibody after all. I will try your idea of refreshing the SuperSignal Pico when this happens in future.

I am satisfed with the antibody and do not believe that I will need to take you up on your offer of a repacement.

Thanks for your help in troubleshooting this problem!

BTW, we just got box of Fairytale Brownies from Eve! Thanks a bunch! You guys are great!

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Answer

Hi, glad to be of help, and Eve is REALLY glad you liked the brownies!

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Question

Thanks for your help today. Yes, we have used the same primary antibody with both anti-mouse secondaries. With the other secondary, as with yours, we occasionally get a too-strong signal that eventually burns out, starting in the middle of the band, and leaves a colored band on the blot. We have learned to incubate that secondary at a slightly higher dilution than the usual 1:2000 (1:3000 - 1:4000) for only 1 hour or so and then the Supersignal West Pico does great. As I told you over the phone, we run only about 10 - 15 ug of protein per lane (unless the protein of interest is weakly expressed). The only time, though, that I have had an issue of weaker bands fading away with the other secondary is when I try to reprobe an old blot. Then I have to actually increase the concentration of the secondary to 1:1000. The supplier doesn't say what the stock concentration of the antibody is, so I cannot compare them directly, but I would guess that theirs is already 10 times less concentrated than yours.

So I will try different dilutions as you suggest, on a dot blot, and compare the two. Should I let the antibody dilutions actually dry onto the membrane? Otherwise wouldn't it just wash off? ( We use nitrocellulose for most everything.) I will get back to you about a replacement antibody, I'd like to do a little digging around first.

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Answer

I have attached our dot blot protocol. However, on second thought, it may not be applicable to a blot of just the secondary, because I am not sure how well the HRP will handle the drying step that is at at the start of the attached protocol, after applying a protein sample to the nitrocellulose. You need the HRP to be active and drying the secondary is not equivalent to what happens when you apply the secondary in a standard WB procedure. (It never drys out in that procedure. On the other hand, the Pierce dot blot of HRP worked. I expect drying is necessary in order to keep it from washing of the blot).

Whatever you decide to do, just let me know whether you would like to try a different secondary or a credit or refund, and I will arrange that for you.

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Question

The secondary anti-mouse IgG antibody ab102458 appears to lose activity after exposure of the blot to the first film, compared to another HRP-conjugated secondary.

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Answer

Thank you for bringing this to our attention. It could very well be that the HRP is the issue, or the antibody:antibody affinity. I am assuming you are using the same primary with both secondaries (the more expensive one from another company and ours).

Without the data from the other secondary, I would say that the fading of chemiluminescent signal is expected. The depletion of substrate is possible, and diluting the antibody might address that, but I would expect that depletion would occur with the other secondary too, if the intensity of the initial signal was comparable to the ab102458 signal.
As I mentioned, if you are interested in trying one of the secondaries in our catalogue that have more sales, I will be happy to send one as a free replacement, for instance ab6789.

Click here (or use the following: https://www.abcam.com/Goat-Mouse-IgG-HL-HRP-ab6789.html).

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