Anti-pro Caspase-12 抗体 (ab8118)

Submit a review Q&A (4)References (2)

Key features and details

  • Rabbit polyclonal to pro Caspase-12
  • Suitable for: IHC-P, WB
  • Reacts with: Mouse, Rat
  • Isotype: IgG

製品の概要

  • 製品名

    Anti-pro Caspase-12 antibody
    pro Caspase-12 一次抗体 製品一覧

  • 製品の詳細

    Rabbit polyclonal to pro Caspase-12

  • 由来種

    Rabbit

  • 特異性

    The antibody only recognizes the prodomain, not the activated form.

  • アプリケーション

    適用あり: IHC-P, WBmore details

  • 種交差性

    交差種: Mouse, Rat
    非交差種: Human

  • 免疫原

    Synthetic peptide corresponding to Mouse pro Caspase-12 aa 2-17 (N terminal).
    Sequence:

    AARRTHERDPIYKIKG


    Database link: Q6UXS9
    (Peptide available as ab8375)

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

製品の特性

  • 製品の状態

    Liquid

  • 保存方法

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

  • バッファー

    Preservative: 0.02% Sodium azide
  • Concentration information loading...

  • 精製度

    Affinity purified

  • 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

Our Abpromise guarantee covers the use of ab8118 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
IHC-P Use a concentration of 10 µg/ml.
WB Use a concentration of 0.5 - 1 µg/ml. Detects a band of approximately 55 kDa.

ターゲット情報

  • 関連性

    Caspase 12 is activated by ER stress, including disruption of ER calcium homeostasis and mediates ER stress-induced apoptosis. Caspase 12 is co-localized to the ER with several proteins that are involved in Alzheimers disease including gamma-secretase presenilin and beta-amyloid precursor protein (APP). Caspase 12 mediates cytotoxicity induced by amyloid-beta. It is ubiquitously expressed in mouse tissue.

  • 細胞内局在

    Endoplasmic reticulum

  • 参照データベース

    • 別名

      • CASP12 antibody
      • CASP12P1 antibody

    画像

    • Western blot - Anti-pro Caspase-12 antibody (ab8118)
      Western blot - Anti-pro Caspase-12 antibody (ab8118)
      All lanes : Anti-pro Caspase-12 antibody (ab8118) at 1 µg/ml

      Lane 1 : Human spleen tissue lysate
      Lane 2 : Mouse spleen tissue lysate
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pro Caspase-12 antibody (ab8118)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pro Caspase-12 antibody (ab8118)

      ab8118 at 10µg/ml staining Caspase-12 in human heart tissue by IHC

    参考文献 (2)

    ab8118 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。

    ab8118 は 2 報の論文で使用されています。

    • Feng SQ  et al. VEGF Antagonism Attenuates Cerebral Ischemia/Reperfusion-Induced Injury via Inhibiting Endoplasmic Reticulum Stress-Mediated Apoptosis. Biol Pharm Bull 42:692-702 (2019). PubMed: 30828041
    • Hauri-Hohl MM  et al. Donor T-cell alloreactivity against host thymic epithelium limits T-cell development after bone marrow transplantation. Blood 109:4080-8 (2007). PubMed: 17213290

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    1-4 of 4 Abreviews or Q&A

    Question

    Thanks for giving the suggestions to the problems the customer has with ab8118. The customer has come back with a couple of questions: 1) Why is it the western blot on the abcam website for ab24582 and ab8118 exactly the same (though they identify different forms of caspase 1)? 2) Before, the customer has bought and used ab8118 with lot number 127548 and was successful ( see attachement picture 1, 55KDa), but later when he bough another ab8118 (lot 132383), the results was not clean (see attachment picture 2) and the major band is not the target protein. Why is that? I look forward to your reply. Thanks

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    Answer

    Thank you for your e-mail. Did you mean ab24852, rather than 24582? The antibodies are against caspase 12, not caspase 1. Can you please confirm that the customer is looking for caspase 12 and not caspase 1? I compared the two sheets of ab24852 and ab8118 and am asking my colleagues for an explanation of this important point. The two antibodies come from different suppliers and therefore we need to get in touch with them and find out why they can supply different products with the same image. It seems that it is the same product to me but I will confirm this. We have already noticed that ab8118 is recommended for human while ab24852 is not. Is the customer using human samples? Thank you for the customer's images. In the first attachment (1) there are two membranes. Is the top membrane the one probed with ab8118? Did it give other bands too? Is the bottom membrane the same? The two lots come from the same master stock 0302 so should give the same quality of bands. Is the customer using the same samples with the two lots or are they different? I look forward to hearing how the customer gets on with the recommended changes and more details on the blot and species of samples.

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    Question

    1. Order details: * Batch number: 127548 * Abcam order or Purchase order number: ab8118 * Antibody storage conditions (temperature/reconstitution etc):aliquot at -20C 2. Please describe the problem (high background, wrong band size, more bands, no band etc). Weak signal on described molecular weight but significant signals on the other site; no correlation pattern with active form . 3. On what material are you testing the antibody in WB? * Species: Rat * Cell extract or Nuclear extract: Cell extract * Purified protein or Recombinant protein:purified trotein 3. The lysate * How much protein was loaded:30 ug * What lysis buffer was used:PBS * What protease inhibitors were used:protease inhibitor cocktail setI.(caltiochem) * What loading buffer was used: SDS gel-loading buffer * Did you heat the samples: temperature and time:100 C , 5 min 4. Electrophoresis/Gel conditions/ Transfer conditions * Reducing or non reducing gel: Reducing gel * Gel percentage : 12 % * Transfer conditions: 90 volts , 90 min 5. Blocking conditions * Buffer: PBST * Blocking agent: milk, BSA, serum, what percentage: 3 % non-fat milk * Incubation time: one hour * Incubation temperature: room temperature 6. Primary Antibody * Specification (in which species was it raised against): rabbit anti-rat * At what dilution(s) have you tested this antibody: 1:1000 * What dilution buffer was used: 3% non-fat milk in PBST * Incubation time: overnight * Incubation temperature: 4 C * What washing steps were done:10 min, three times 7. Secondary Antibody * Specification (in which species was it raised against)? goat anti-rabbit * At what dilution(s) have you tested this antibody:1;3000 * Incubation time: two hours * Wash steps: 10 min, three times * Do you know whether the problems you are experiencing come from the secondary? We do not think the problems comeing from the secondary antibody. 8. Detection method ECl, ECl+, other detection method: ECL 9. Background bands * Have you eliminated the possibility that any background bands could be due to the secondary antibody? (Run a "No primary" control): No, but it would not be the factor. * Is the blocking step sufficient? Yes * Are your washing steps sufficiently stringent? (Multiple short washes are more effective than fewer longer wash steps) Yes * At what size are the bands migrating? Could they be degradation products of your target? * Please provide an image of your blot (as an e-mail attachment, a faxed image is not sufficient) Fine 11. Did you apply positive and negative controls along with the samples? Please specify. Yes. 0. Optimization attempts * How many times have you tried the Western? Exceed ten times * Do you obtain the same results every time e.g. are background bands always in the same place? Yes.

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    Answer

    I'm sorry to hear your customer is experiencing problems with ab8118. The customer mentions that he/she provided images however these did not reach us, so I cannot see the membrane pattern with this antibody I'm afraid. It is very unclear what type of tissue was used and I would like to recommend a positive control of Human or Mouse spleen tissue lysate to make sure that the low signal at the correct band size is not due to low levels of the protein in the customer's samples. The protein is located at the ER and thus needs to be extracted with a strong detergent, a PBS buffer will therefore not be sufficient and I recommend to use a RIPA buffer for example, as the NP40 will lyse the cells very well. It is also very important to make sure that the right amount of protease inhibitors is added, and that those are fresh as they are often responsible for multiple bands unfortunately. The customer should also try a 1:500 dilution of the antibody, as recommended on the datasheet, to enhance his signal. I think an Ecl+ kit or more sensitive kits are more appropriate than Ecl, which is less performant. Regarding the problem of background bands, I would recommend to use 5% BSA for 1hr in TBST (not PBST) (Tween 0.1%) and to try to incubate the antibody in TBST only (and to try also with 1-2% BSA) (typically we incubate in TBST only but with some samples a little BSA in the antibody dilution buffer reduces the background). As the secondary antibody has not been tested alone I would suggest to try this, just to make sure that it is indeed performing well. I hope these recommendations will help. Please do not hesitate to contact me again should the customer still experience problems with those changes,

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    Question

    How do I distinguish between activated/non activated forms? Is there an increase in the amount of caspase 12 upon activation of the cells or are the activated and non-activated forms different is size? Is a positive control supplied with the antibody?

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    Answer

    This antibody is raised against the N-terminal sequence AARRTHERDPIYKIKG. If this sequence forms part of the pro-peptide, then this antibody will be selective for the non-activated form. The blocking peptide for this antibody is available separately, ab8375.

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    Question

    I am interested in purchasing your caspase 12 antibody. Do either of the antibodies listed on your webpage recognize the procaspase 12 and the cleaved form? Your site only mentions that one band (43 KDa) is recognized on western blot. I would like to use the antibody to determine if caspase 12 is active.

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    Answer

    These two antibodies ab8118 and ab8117 recognize the pre-activated form of caspase 12, however we have not tested for the activated/cleaved form.

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