Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042) is a rabbit monoclonal antibody detecting Cleaved Caspase-3 in Western Blot, ICC/IF. Suitable for Human,.
- KO validated for confirmed specificity
- Biophysical QC for unrivalled batch-batch consistency
- Over 610 publications
- Trusted since 2006
Images
![Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (AB32042), expandable thumbnail](https://content.abcam.com/products/images/cleaved-caspase-3-antibody-e83-77-ab32042--western-blot-img16887.jpg/_jcr_content/renditions/webp-475.webp "Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (AB32042)")
![Immunocytochemistry/ Immunofluorescence - Anti-Cleaved Caspase-3 antibody [E83-77] (AB32042), expandable thumbnail](https://content.abcam.com/products/images/cleaved-caspase-3-antibody-e83-77-ab32042--immunocytochemistry-immunofluorescence-img92858.jpg/_jcr_content/renditions/webp-475.webp "Immunocytochemistry/ Immunofluorescence - Anti-Cleaved Caspase-3 antibody [E83-77] (AB32042)")
![Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (AB32042), expandable thumbnail](https://content.abcam.com/products/images/cleaved-caspase-3-antibody-e83-77-ab32042--western-blot-img62178.jpg/_jcr_content/renditions/webp-475.webp "Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (AB32042)")
![Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (AB32042), expandable thumbnail](https://content.abcam.com/products/images/cleaved-caspase-3-antibody-e83-77-ab32042--western-blot-img56999.jpg/_jcr_content/renditions/webp-475.webp "Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (AB32042)")
![Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (AB32042), expandable thumbnail](https://content.abcam.com/products/images/cleaved-caspase-3-antibody-e83-77-ab32042--western-blot-img416795.jpg/_jcr_content/renditions/webp-475.webp "Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (AB32042)")
Publications
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- PeerJ 11:e165932023Inhibition of hepatocellular carcinoma growth modulation of the miR-221/SOX11 axis by curcumin and berberine.Applications:Unspecified applicationReactive species:Unspecified reactive speciesSheng Li,Xiaoliang Cai,Liang Chen,Manbian Lin,Ziqi Zhu,Huihuang Xiao,Pingping Nie,Quanwen Chen,Xiaoyu YangPubMed 38084140
- Nature communications 14:78562023A click chemistry-mediated all-peptide cell printing hydrogel platform for diabetic wound healing.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJinjian Huang,Rong Yang,Jiao Jiao,Ze Li,Penghui Wang,Ye Liu,Sicheng Li,Canwen Chen,Zongan Li,Guiwen Qu,Kang Chen,Xiuwen Wu,Bo Chi,Jianan RenPubMed 38030636
- Analytical cellular pathology (Amsterdam) 2023:11082802023Depletion of lncRNA MEG3 Ameliorates Imatinib-Induced Injury of Cardiomyocytes via Regulating miR-129-5p/HMGB1 Axis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesPeng Tang,Jinjian Zhou,Huagang Liu,Shenglan Mei,Kai Wang,Hao MingPubMed 38028435
- American journal of cancer research 13:5577-55892023Detection of CSTF2 by nano fluorescent probe and its correlation with malignant biological characteristics in liver cancer.Applications:Unspecified applicationReactive species:Unspecified reactive speciesZhongmin Jiang,Xiaozhi Liu,Yongqing Deng, Duojierenqing, Renqingcuo,Wenhan Wu,Wei Meng,Zhijiang ShaoPubMed 38058835
- Signal transduction and targeted therapy 8:4282023Microcolin H, a novel autophagy inducer, exerts potent antitumour activity by targeting PITPα/β.Applications:Unspecified applicationReactive species:Unspecified reactive speciesHange Yang,Xiaowei Zhang,Cong Wang,Hailong Zhang,Juan Yi,Kun Wang,Yanzhe Hou,Peihong Ji,Xiaojie Jin,Chenghao Li,Min Zhang,Shan Huang,Haoyuan Jia,Kuan Hu,Lingyun Mou,Rui WangPubMed 37963877
- Molecular and cellular biochemistry :2023MIA3 promotes the degradation of GSH (glutathione) by binding to CHAC1, thereby promoting the progression of hepatocellular carcinoma.Applications:Unspecified applicationReactive species:Unspecified reactive speciesZhou Wanbiao,Man Jing,Zuo Shi,Chen Tengxiang,Zhao Xueke,Li HaiyangPubMed 37948019
- Molecular medicine reports 28:2023IGF‑1 inhibits palmitic acid‑induced mitochondrial apoptosis in macrophages.Applications:Unspecified applicationReactive species:Unspecified reactive speciesWanying Tang,Ming Zhang,Yu Wang,Dan Ma,Mi Hu,Yangkai Zhang,Huiling Lin,Weiwei Jiang,Yuxin Ouyang,Liping Jiang,Pingping He,Guojun Zhao,Xinping OuyangPubMed 37921069
- Allergologia et immunopathologia 51:1-72023Kaempferitrin alleviates LPS-induced septic acute lung injury in mice through downregulating NF-κB pathway.Applications:Unspecified applicationReactive species:Unspecified reactive speciesXiaoli Zhu,Yongyue Pan,Xin Xu,Jing XuPubMed 37937489
- Nature communications 14:69052023An autocatalytic multicomponent DNAzyme nanomachine for tumor-specific photothermal therapy sensitization in pancreatic cancer.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJiaqi Yan,Xiaodong Ma,Danna Liang,Meixin Ran,Dongdong Zheng,Xiaodong Chen,Shichong Zhou,Weijian Sun,Xian Shen,Hongbo ZhangPubMed 37903795
- BMC complementary medicine and therapies 23:3812023Afzelin induces immunogenic cell death against lung cancer by targeting NQO2.Applications:Unspecified applicationReactive species:Unspecified reactive speciesLei Xia,Xiaoqing Xu,Meijun Li,Xinyue Zhang,Fang CaoPubMed 37891619
Key facts
- Isotype
- IgG
- Host species
- Rabbit
- Storage buffer
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 50% Glycerol (glycerin, glycerine), 49% PBS, 0.05% BSA- Form
- Liquid
- Clonality
- Monoclonal
Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
Abcam Recommends
Reactivity data
Select an application| Flow Cyt | IP | WB | IHC-P | ICC/IF | |
|---|---|---|---|---|---|
| Human | Not recommended | Not recommended | Tested | Not recommended | Tested |
| Recombinant fragment | Not recommended | Not recommended | Not recommended | Not recommended | Not recommended |
| Recombinant fragment - Human | Not recommended | Not recommended | Tested | Not recommended | Not recommended |
Flow Cyt
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Recombinant fragment, Human, Recombinant fragment - Human | Dilution info - | Notes - |
IP
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Human, Recombinant fragment, Recombinant fragment - Human | Dilution info - | Notes - |
WB
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Recombinant fragment - Human | Dilution info 1/500 | Notes The pro-caspase-3 is cleaved only when apoptosis event occurs. So, in order to detect active Caspase-3, we strongly suggest to induce your samples into apoptotic pathway. |
Species Human | Dilution info 1/500 | Notes The pro-caspase-3 is cleaved only when apoptosis event occurs. So, in order to detect active Caspase-3, we strongly suggest to induce your samples into apoptotic pathway. |
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Recombinant fragment | Dilution info - | Notes - |
IHC-P
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Human, Recombinant fragment, Recombinant fragment - Human | Dilution info - | Notes - |
ICC/IF
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info 1/100 - 1/250 | Notes - |
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Recombinant fragment, Recombinant fragment - Human | Dilution info - | Notes - |
Associated Products
Select an associated product type
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Target data
Function
Thiol protease that acts as a major effector caspase involved in the execution phase of apoptosis (PubMed:18723680, PubMed:20566630, PubMed:23650375, PubMed:35338844, PubMed:35446120, PubMed:7596430). Following cleavage and activation by initiator caspases (CASP8, CASP9 and/or CASP10), mediates execution of apoptosis by catalyzing cleavage of many proteins (PubMed:18723680, PubMed:20566630, PubMed:23650375, PubMed:7596430). At the onset of apoptosis, it proteolytically cleaves poly(ADP-ribose) polymerase PARP1 at a '216-Asp-|-Gly-217' bond (PubMed:10497198, PubMed:16374543, PubMed:7596430, PubMed:7774019). Cleaves and activates sterol regulatory element binding proteins (SREBPs) between the basic helix-loop-helix leucine zipper domain and the membrane attachment domain (By similarity). Cleaves and activates caspase-6, -7 and -9 (CASP6, CASP7 and CASP9, respectively) (PubMed:7596430). Cleaves and inactivates interleukin-18 (IL18) (PubMed:37993714, PubMed:9334240). Involved in the cleavage of huntingtin (PubMed:8696339). Triggers cell adhesion in sympathetic neurons through RET cleavage (PubMed:21357690). Cleaves and inhibits serine/threonine-protein kinase AKT1 in response to oxidative stress (PubMed:23152800). Acts as an inhibitor of type I interferon production during virus-induced apoptosis by mediating cleavage of antiviral proteins CGAS, IRF3 and MAVS, thereby preventing cytokine overproduction (PubMed:30878284). Also involved in pyroptosis by mediating cleavage and activation of gasdermin-E (GSDME) (PubMed:35338844, PubMed:35446120). Cleaves XRCC4 and phospholipid scramblase proteins XKR4, XKR8 and XKR9, leading to promote phosphatidylserine exposure on apoptotic cell surface (PubMed:23845944, PubMed:33725486). Cleaves BIRC6 following inhibition of BIRC6-caspase binding by DIABLO/SMAC (PubMed:36758104, PubMed:36758106).
Alternative names
CPP32, CASP3, Caspase-3, CASP-3, Apopain, Cysteine protease CPP32, Protein Yama, SREBP cleavage activity 1, CPP-32, SCA-1
Recommended products
Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042) is a rabbit monoclonal antibody detecting Cleaved Caspase-3 in Western Blot, ICC/IF. Suitable for Human,.
- KO validated for confirmed specificity
- Biophysical QC for unrivalled batch-batch consistency
- Over 610 publications
- Trusted since 2006
Key facts
- Isotype
- IgG
- Host species
- Rabbit
- Storage buffer
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 50% Glycerol (glycerin, glycerine), 49% PBS, 0.05% BSA- Form
- Liquid
- Clonality
- Monoclonal
- Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
- Clone number
- E83-77
- Purification technique
- Affinity purification Protein A
- Specificity
This antibody is more sensitive for the detection of cleaved caspase-3 than for pro-caspase-3. For better observation of pro-caspase-3, we recommend ab32150.
PubMedID 19789217 describes the detection of human cells injected into mice. The pro-caspase-3 is cleaved only when apoptosis event occurs. So, in order to detect active Caspase-3, we strongly suggest to induce your samples into apoptotic pathway.
- Concentration
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- -20°C
- Aliquoting information
- Upon delivery aliquot
- Storage information
- Avoid freeze / thaw cycle
Notes
Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in ICC/IF and WB.
Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042) was first used in a scientific publication in 1994 and has been cited over 610 times in peer reviewed journals. It's performance in Western Blot in human samplesis trusted by the scientific community.
Abcam's high quality manufacturing and validation processes ensure Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.
The specificity of Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042) has been confirmed by Western Blot testing in Cleaved Caspase-3 knockout HAP1 cells.
Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042) has 25 independent reviews from customers.
Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042) specifically detects Cleaved Caspase-3 (UniProt ID: P42574; Molecular weight: 17kDa) and is sold in a convenient trial size to enable initial testing (20 µL) and larger sizes for subsequent scaling up experiments (100 µL and 1 mL).
Conjugation-ready, carrier free format available for antibody clone E83-77 - Anti-Cleaved Caspase-3 antibody [E83-77] - BSA and Azide free ab208003.
Cleaved Caspase-3 is a crucial marker of apoptosis, the programmed cell death process. It is the activated form of Caspase-3, resulting from proteolytic cleavage. Cleaved Caspase-3 plays a vital role in executing apoptosis by cleaving various cellular substrates, leading to cell dismantling. Researchers use Cleaved Caspase-3 as a reliable indicator of apoptosis in studies of cancer, neurodegenerative diseases and other conditions involving cell death. Its detection is essential for understanding the mechanisms of apoptosis and developing targeted therapies.
In cancer research, cleaved Caspase-3 is used to assess the effectiveness of anti-cancer therapies that induce apoptosis in tumor cells. Elevated levels of cleaved Caspase-3 have been associated with better prognosis in certain cancers, such as oral tongue squamous cell carcinoma. Additionally, cleaved Caspase-3 has been implicated in non-apoptotic roles, such as promoting angiogenesis and chemotherapy resistance.
Neurodegenerative Diseases: In neurodegenerative diseases like Alzheimer's (AD) and Parkinson's (PD) disease, cleaved Caspase-3 is involved in the apoptotic pathways that lead to neuronal cell death. For instance, caspase-3-cleaved Tau aggregates more easily than full-length Tau, contributing to the pathology of Alzheimer's disease.
Understanding the role of cleaved Caspase-3 in these diseases helps researchers develop targeted therapies to mitigate neuronal loss and disease progression.
Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
What are the advantages of a recombinant monoclonal antibody?
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 batch production
For more information, read more on recombinant antibodies.
Species reactivity
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species.
Please contact us for more information.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
The Cleaved Caspase-3 also known as active caspase 3 plays an important role in the execution phase of cell apoptosis. The caspase 3 molecule has a molecular weight of approximately 32 kDa. It undergoes cleavage into subunits that are important for its activation marking its conversion into the cleaved form. Cleaved caspase is widely expressed in various tissues with a notable presence in the cytoplasm of cells undergoing apoptotic processes. Cleavage of caspase 3 results in the formation of a functional enzyme that participates in the degradation of cellular components during apoptosis.
- Biological function summary
Caspase 3 acts as an executioner enzyme in the process of programmed cell death. Its cleavage from the inactive zymogen form into the active form triggers apoptotic pathways ensuring the removal of damaged or unnecessary cells. Cleaved caspase operates within a proteolytic cascade often in conjunction with other caspases such as caspase 9. Once activated caspase 3 targets and cleaves specific cellular substrates contributing to the characteristic morphological changes and DNA fragmentation associated with apoptosis.
- Pathways
Cleaved caspase 3 is significant within the intrinsic and extrinsic apoptotic pathways. It acts downstream of initiator caspases such as caspase 9 in the intrinsic pathway where its activation involves mitochondrial signals. In the extrinsic pathway it closely associates with caspase 8 mediating cell death signals from extracellular triggers. These pathways ensure that cleaved caspase 3 fulfills its role as a critical mediator of apoptosis linking upstream death signals to cellular dismantling during the apoptosis process.
- Associated diseases and disorders
Cleaved caspase 3 connects to conditions such as cancer and neurodegenerative diseases. In cancer dysregulation of caspase 3 activity may lead to evasion of apoptosis facilitating tumor progression. Therapeutic approaches aim to restore its apoptotic functions in cancer cells. In neurodegenerative diseases like Alzheimer's disease abnormal activation of cleaved caspase 3 contributes to neuronal loss. It interacts with proteins like amyloid precursor protein promoting the pathological changes associated with this disorder. Understanding these interactions provides insights into potential therapeutic avenues.
Product promise
Tested
We have tested this species and application combination and it works. It is covered by our product promise.
Expected
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
Predicted
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
Not recommended
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
5 product images
![Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042), expandable thumbnail](https://content.abcam.com/products/images/cleaved-caspase-3-antibody-e83-77-ab32042--western-blot-img16887.jpg "Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)")
Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)
Lane 1: Wild type HAP1 + DMSO for 24 hours, whole cell lysate (20 μg)
Lane 2: Wild type HAP1 + 2uM Staurosporine (Staurosporine (DMSO solution), protein kinase inhibitor ab146588) for 24 hours, whole cell lysate (20 μg)
Lane 3: HAP1 CASP3 KO + DMSO for 24 hours, whole cell lysate (20 μg)
Lane 4: HAP1 CASP3 KO + 2uM Staurosporine (Staurosporine (DMSO solution), protein kinase inhibitor ab146588) for 24 hours, whole cell lysate (20 μg)
Lane 5: HeLa + DMSO for 24 hours, whole cell lysate (20 μg)
Lane 6: HeLa + 2uM Staurosporine (Staurosporine (DMSO solution), protein kinase inhibitor ab146588) for 24 hours, whole cell lysate (20 μg)
Lanes 1 - 6: Merged signal (red and green). Green - ab32042 observed at 17 kDa. Red - loading control, Anti-Vinculin antibody [VIN-54] ab130007, observed at 130 kDa.ab32042 was shown to specifically react with CASP3 (Caspase-3) when CASP3 (Caspase-3) knockout samples were used. HAP1 wild-type and CASP3 (Caspase-3) knockout samples were subjected to SDS-PAGE. ab32042 and Anti-Vinculin antibody [VIN-54] ab130007 (Mouse anti vinculin loading control) were incubated overnight at 4°C at 500 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Cells were grown to confluency prior to treatment.
All lanes: Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)
Predicted band size: 32 kDa
![Immunocytochemistry/ Immunofluorescence - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042), expandable thumbnail](https://content.abcam.com/products/images/cleaved-caspase-3-antibody-e83-77-ab32042--immunocytochemistry-immunofluorescence-img92858.jpg "Immunocytochemistry/ Immunofluorescence - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)")
Image from Zhu ZX et al., PLoS One. 2015;10(12):e0144495. Fig 7.; doi: 10.1371/journal.pone.0144495. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/Immunocytochemistry/ Immunofluorescence - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)
High-glucose induces apoptosis in human Vascular endothelial cells (VECs).Apoptotic responses in VEC were analyzed by detection of cleaved-Caspase-3 immunofluorescence using ab32042. Cells were treated with low glucose (LG) or high glucose (HG) for 72 hours before treated with 100 ng/mL bFGF, 1 μM sp600125 (sp), or 1 μM U0126 (U) or 10 μM MnTmPyP for 1 hour. Bar = 100 μm.
![Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042), expandable thumbnail](https://content.abcam.com/products/images/cleaved-caspase-3-antibody-e83-77-ab32042--western-blot-img62178.jpg "Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)")
Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)
All lanes: Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042) at 1/500 dilution
Lane 1: HeLa Whole Cell Lysate (2 uM Staurosporine, 4Hr) at 20 µg
Lane 2: HeLa Whole Cell Lysate (untreated) at 20 µg
Lane 3: Cleaved Caspase 3 (recombinant protein) at 0.1 µg
SecondaryAll lanes: 800CW Goat Anti-Rabbit IgG at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 32 kDa
Observed band size: 17 kDa
![Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042), expandable thumbnail](https://content.abcam.com/products/images/cleaved-caspase-3-antibody-e83-77-ab32042--western-blot-img56999.jpg "Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)")
Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)
Lanes 1 - 2: anti Pro Caspase 3 at 1/10000 dilution
Lanes 3 - 4: Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042) at 1/500 dilution
Lane 1: Jurkat (human T cell leukemia cell line from peripheral blood) cell lysate
Lanes 2 and 4: Jurkat cell lysate + Camptothecin
Lane 3: Jurkat cell lysate
Predicted band size: 32 kDa
Observed band size: 17 kDa, 30 kDa
![Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042), expandable thumbnail](https://content.abcam.com/products/images/cleaved-caspase-3-antibody-e83-77-ab32042--western-blot-img416795.jpg "Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)")
Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)
Cleaved Caspase-3 Western blot staining using rabbit Anti-Cleaved Caspase-3 antibody
Western blot: Anti-CASP3 antibody [E83-77] (ab32042) staining at 1/500 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32042 was shown to bind specifically to CASP3. A band was observed at 16/28 kDa in treated wild-type HeLa cell lysates with no signal observed at this size in CASP3 knockout cell line. To generate this image, wild-type and CASP3 knockout HeLa 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.
All lanes: Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042) at 1/500 dilution
Lane 1: Wild-type HeLa Treated Staurosporine (2uM, 4h) cell lysate at 20 µg
Lane 2: CASP3 knockout HeLa Treated Staurosporine (2uM, 4h) cell lysate at 20 µg
Lane 3: Wild-type HeLa Vehicle Control Staurosporine (0uM, 4h) cell lysate, at 20 µg
Lane 4: CASP3 knockout HeLa Vehicle Control Staurosporine (0uM, 4h) cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 32 kDa
Observed band size: 16 kDa, 28 kDa
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