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遺伝子ノックアウト細胞株 検証済リコンビナントRabMAb

Anti-Caspase-3 抗体 [EPR18297] (ab184787)

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Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787)
  • Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787)
  • Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787)
  • Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787)
  • Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-3 antibody [EPR18297] (ab184787)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-3 antibody [EPR18297] (ab184787)
  • Immunoprecipitation - Anti-Caspase-3 antibody [EPR18297] (ab184787)
  • Anti-Caspase-3 antibody [EPR18297] (ab184787)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18297] to Caspase-3
  • Suitable for: WB, IHC-P, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

製品の概要

  • 製品名

    Anti-Caspase-3 antibody [EPR18297]
    Caspase-3 一次抗体 製品一覧

  • 製品の詳細

    Rabbit monoclonal [EPR18297] to Caspase-3

  • 由来種

    Rabbit

  • 特異性

    This antibody recognizes pro-Caspase 3 and potentially cross reacts with active caspases after apoptosis has been induced in wildtype cells and not Caspase 3 knockout cells

  • アプリケーション

    適用あり: WB, IHC-P, IPmore details

  • 種交差性

    交差種: Mouse, Rat, Human

  • 免疫原

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • ポジティブ･コントロール

    • WB: Wild-type HAP1 treated Staurosporine (2 uM, 4h) and Vehicle Control Staurosporine (0 uM, 4h), Wild-type HeLa treated Staurosporine (2 uM, 4h) and Vehicle Control Staurosporine (0 uM, 4h), untreated NIH/3T3 and treated with 1µM Staurosporine for 4 hours, untreated Jurkat and treated with 1uM Staurosporine for 4 hours, mouse and rat brain tissue lysates. Human brain, heart and liver tissue. IHC-P: Human tonsil and cervical cancer tissues. IP: HeLa lysate treated with 1uM Staurosporine for 4 hours.

  • 特記事項

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

製品の特性

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ab184787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
WB (2)
1/2000. Detects a band of approximately 32, 17 kDa (predicted molecular weight: 32 kDa).

The 17 kDa band is the active form of the cleaved caspase 3 (subunit p17).
ab184787 recognizes pro-Caspase 3 and unable to detect the active caspases after induction in mouse and rat samples.
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
1/80.
特記事項
WB
1/2000. Detects a band of approximately 32, 17 kDa (predicted molecular weight: 32 kDa).

The 17 kDa band is the active form of the cleaved caspase 3 (subunit p17).
ab184787 recognizes pro-Caspase 3 and unable to detect the active caspases after induction in mouse and rat samples.
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
1/80.

ターゲット情報

  • 機能

    Involved in the activation cascade of caspases responsible for apoptosis execution. At the onset of apoptosis it proteolytically cleaves poly(ADP-ribose) polymerase (PARP) at a '216-Asp-
    -Gly-217' bond. Cleaves and activates sterol regulatory element binding proteins (SREBPs) between the basic helix-loop-helix leucine zipper domain and the membrane attachment domain. Cleaves and activates caspase-6, -7 and -9. Involved in the cleavage of huntingtin.

  • 組織特異性

    Highly expressed in lung, spleen, heart, liver and kidney. Moderate levels in brain and skeletal muscle, and low in testis. Also found in many cell lines, highest expression in cells of the immune system.

  • 配列類似性

    Belongs to the peptidase C14A family.

  • 翻訳後修飾

    Cleavage by granzyme B, caspase-6, caspase-8 and caspase-10 generates the two active subunits. Additional processing of the propeptides is likely due to the autocatalytic activity of the activated protease. Active heterodimers between the small subunit of caspase-7 protease and the large subunit of caspase-3 also occur and vice versa.
    S-nitrosylated on its catalytic site cysteine in unstimulated human cell lines and denitrosylated upon activation of the Fas apoptotic pathway, associated with an increase in intracellular caspase activity. Fas therefore activates caspase-3 not only by inducing the cleavage of the caspase zymogen to its active subunits, but also by stimulating the denitrosylation of its active site thiol.

  • 細胞内局在

    Cytoplasm.
  • Information by UniProt

  • 参照データベース

    see all

  • 別名

    • A830040C14Rik antibody
    • Apopain antibody
    • CASP 3 antibody
    • CASP-3 antibody
    • CASP3 antibody
    • CASP3_HUMAN antibody
    • Casp3a antibody
    • Caspase 3 antibody
    • Caspase 3, apoptosis-related cysteine peptidase antibody
    • Caspase 3, apoptosis-related cysteine protease antibody
    • Caspase 3, apoptosis-related cysteine protease a antibody
    • Caspase-3 subunit p12 antibody
    • Caspase3 antibody
    • CC3 antibody
    • CPP 32 antibody
    • CPP-32 antibody
    • CPP32 antibody
    • CPP32B antibody
    • Cysteine protease CPP32 antibody
    • EC 3.4.22.56 antibody
    • ICE3 antibody
    • LICE antibody
    • mldy antibody
    • OTTHUMP00000165052 antibody
    • OTTHUMP00000165053 antibody
    • OTTHUMP00000165054 antibody
    • PARP cleavage protease antibody
    • Procaspase3 antibody
    • Protein Yama antibody
    • SCA 1 antibody
    • SCA-1 antibody
    • SCA1 antibody
    • SREBP cleavage activity 1 antibody
    • Yama antibody
    • Yama protein antibody
    see all

画像

  • Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787)
    Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787)
    All lanes : Anti-Caspase-3 antibody [EPR18297] (ab184787) at 1/1000 dilution

    Lane 1 : Mouse Alzheimer's disease brain tissue lysate
    Lane 2 : Mouse brain cancer tissue lysate
    Lane 3 : Mouse hippocampus tissue lysate
    Lane 4 : Mouse spinal cord tissue lysate
    Lane 5 : Mouse cerebellum tissue lysate
    Lane 6 : Mouse cerebral cortex tissue lysate
    Lane 7 : Mouse hypothalamus tissue lysate
    Lane 8 : Mouse heart tissue lysate
    Lane 9 : Mouse liver tissue lysate
    Lane 10 : Human brain tissue lysate
    Lane 11 : Human liver tissue lysate
    Lane 12 : Human hypothalamus tissue lysate
    Lane 13 : Human heart tissue lysate
    Lane 14 : Human cerebellum tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

    Predicted band size: 32 kDa
    Observed band size: 27-32 kDa why is the actual band size different from the predicted?


    Exposure time: 10 seconds


    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    ab181602 was used as GAPDH loading control.

    Bands between 27-32kDa represent cleavage of the procaspase at D9 and D28, respectively (PMID: 14567691)

  • Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787)
    Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787)
    All lanes : Anti-Caspase-3 antibody [EPR18297] (ab184787) at 1/1000 dilution

    Lane 1 : Rat brain tissue lysate
    Lane 2 : Rat hippocampus tissue lysate
    Lane 3 : Rat spinal cord tissue lysate
    Lane 4 : Rat cerebellum tissue lysate
    Lane 5 : Rat cerebral cortex tissue lysate
    Lane 6 : Rat hypothalamus tissue lysate
    Lane 7 : Rat heart tissue lysate
    Lane 8 : Rat liver tissue lysate
    Lane 9 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

    Predicted band size: 32 kDa
    Observed band size: 27-32 kDa why is the actual band size different from the predicted?


    Exposure time: 20 seconds


    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    ab181602 was used as GAPDH loading control.

    Bands between 27-32kDa represent cleavage of the procaspase at D9 and D28, respectively (PMID: 14567691)

  • Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787)
    Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787)
    All lanes : Anti-Caspase-3 antibody [EPR18297] (ab184787) at 1/2000 dilution

    Lane 1 : Wild-type HAP1 Treated Staurosporine (2 uM, 4h) cell lysate
    Lane 2 : CASP3 knockout HAP1 Treated Staurosporine (2 uM, 4h) cell lysate
    Lane 3 : Wild-type HAP1 Vehicle Control Staurosporine (0 uM, 4h) cell lysate
    Lane 4 : CASP3 knockout HAP1 Vehicle Control Staurosporine (0 uM, 4h) cell lysate
    Lane 5 : Wild-type HeLa Treated Staurosporine (2 uM, 4h) cell lysate
    Lane 6 : CASP3 knockout HeLa Treated Staurosporine (2 uM, 4h) cell lysate
    Lane 7 : Wild-type HeLa Vehicle Control Staurosporine (0 uM, 4h) cell lysate
    Lane 8 : CASP3 knockout HeLa Vehicle Control Staurosporine (0 uM, 4h) cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

    Performed under reducing conditions.

    Predicted band size: 32 kDa
    Observed band size: 35 kDa why is the actual band size different from the predicted?



    Anti-CASP3 antibody [EPR18297] (ab184787) staining at 1/2000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab184787 was shown to bind specifically to CASP3. A band was observed at 35 kDa in treated wild-type HAP1 and HeLa cell lysates with no signal observed at this size in CASP3 knockout HAP1 cell line and a band at a lower molecular weight in the CAPS3 knockout HeLa cell line ab255370 (knockout cell lysate ab263779) which cannot be cleaved to active CASP3. To generate this image, wild-type and CASP3 knockout HAP1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  • Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787)
    Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787)
    All lanes : Anti-Caspase-3 antibody [EPR18297] (ab184787) at 0.7 µg/ml

    Lane 1 : Untreated NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
    Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) treated with 1µM Staurosporine for 4 hours whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 32 kDa
    Observed band size: 32 kDa



    Blocking and diluting buffer: 5% NFDM/TBST.

    ab184787 recognizes pro-Caspase 3 and unable to detect the active caspases after induction in mouse and rat samples.

  • Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787)
    Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787)
    All lanes : Anti-Caspase-3 antibody [EPR18297] (ab184787) at 1/2000 dilution

    Lane 1 : Untreated Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysates
    Lane 2 : Jurkat whole cell lysates treated with 1uM staurosporine for 4 hours

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 32 kDa
    Observed band size: 17,32 kDa why is the actual band size different from the predicted?


    Exposure time: 1 minute


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Specificity: interacts with full length pro-Caspase 3 and the p17 subunit.

    The Caspase-3 precursor is first cleaved between D175 and S176 to produce the p11 subunit and p20 fragment. Subsequently, the p20 fragment is cleaved between D28 and S29 to generate the p17 subunit (Proc. Natl. Acad. Sci. USA. 93, 7464-7469 - PMID:8755496).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-3 antibody [EPR18297] (ab184787)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-3 antibody [EPR18297] (ab184787)

    Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling active and pro Caspase 3 with ab184787 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus and cytoplasm staining on lymphocytes of tonsil is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-3 antibody [EPR18297] (ab184787)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-3 antibody [EPR18297] (ab184787)

    Immunohistochemical analysis of paraffin-embedded Human cervical cancer tissue labeling active and pro Caspase 3 with ab184787 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus and cytoplasm staining on tumor cells of Human cervix cancer is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunoprecipitation - Anti-Caspase-3 antibody [EPR18297] (ab184787)
    Immunoprecipitation - Anti-Caspase-3 antibody [EPR18297] (ab184787)

    active and pro Caspase 3 was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate treated with 1uM staurosporine for 4 hours with ab184787 at 1/80 dilution. Western blot was performed from the immunoprecipitate using ab184787 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/1500 dilution.

    Lane 1: HeLa whole cell lysate treated with 1uM staurosporine for 4 hours 10 µg (Input).

    Lane 2: ab184787 IP in HeLa whole cell lysate treated with 1uM staurosporine for 4 hours.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab184787 in HeLa whole cell lysate treated with 1uM staurosporine for 4 hours.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 minutes.

  • Anti-Caspase-3 antibody [EPR18297] (ab184787)
    Anti-Caspase-3 antibody [EPR18297] (ab184787)

参考文献 (223)

ab184787 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。

ab184787 は 223 報の論文で使用されています。

  • Wu C  et al. Mapk7 deletion in chondrocytes causes vertebral defects by reducing MEF2C/PTEN/AKT signaling. Genes Dis 11:964-977 (2024). WB ; Mouse . PubMed: 37692479
  • Jiang D  et al. microRNA-105-5p protects against chondrocyte injury, extracellular matrix degradation, and osteoarthritis progression by targeting SPARCL1. Histol Histopathol 39:483-496 (2024). PubMed: 37534667
  • Liao Z  et al. Network pharmacology prediction and experimental verification of Rhubarb-Peach Kernel promoting apoptosis in endometriosis. BMC Complement Med Ther 23:291 (2023). PubMed: 37598188
  • Xiong J  et al. Exosomes from Adipose-Derived Mesenchymal Stem Cells Protect Against Cyclophosphamide-Induced Cardiotoxicity in Rats. Int Heart J 64:935-944 (2023). PubMed: 37778997
  • Li Y  et al. Study of Therapeutic Mechanisms of Puerarin against Sepsis-Induced Myocardial Injury by Integrating Network Pharmacology, Bioinformatics Analysis, and Experimental Validation. Crit Rev Immunol 43:25-42 (2023). PubMed: 37824375
View all Publications for this product

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Western blot abreview for Anti-Caspase-3 antibody [EPR18297]

 Poor
Abreviews
abreview image
Application
Western blot
Sample
Dog Cell lysate - whole cell (melanoma cell line)
Gel Running Conditions
Reduced Denaturing
Loading amount
10 µg
Specification
melanoma cell line
Blocking step
block ace as blocking agent for 50 minute(s) · Concentration: 100% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. 令 中野

Verified customer

投稿 Jun 17 2019

Western blot abreview for Anti-Caspase-3 antibody [EPR18297]

 Good
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Glioma)
Gel Running Conditions
Reduced Denaturing (12.5% gel)
Loading amount
25 µg
Specification
Glioma
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 May 23 2016

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