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Product Info Summary
| SKU: | M00334-6 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Mouse |
| Application: | Flow Cytometry, IHC, WB |
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Product info
Product Name
Anti-Caspase-3 Antibody Picoband® (monoclonal, 8B6)
SKU/Catalog Number
M00334-6
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Caspase-3 Antibody Picoband® (monoclonal, 8B6) catalog # M00334-6. Tested in Flow Cytometry, IHC, WB applications. This antibody reacts with Human. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-Caspase-3 Antibody Picoband® (monoclonal, 8B6) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M00334-6)
Host
Mouse
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Monoclonal
Clone Number
8B6
Isotype
Mouse IgG1
Immunogen
E.coli-derived human Caspase-3 recombinant protein (Position: T67-D175). Human Caspase-3 shares 86% and 90% amino acid (aa) sequence identity with mouse and rat Caspase-3, respectively.
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
M00334-6 is reactive to CASP3 in Human
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500μg/ml.
Observed Molecular Weight
35 kDa
Calculated molecular weight
31.608kDa
Background of Caspase-3
Caspase 3 is a caspase protein which interacts with Survivin, XIAP, CFLAR, Caspase 8, HCLS1, Deleted in Colorectal Cancer, TRAF3 and GroEL. This gene which is located on 4q35 encodes a protein that is a member of the cysteine-aspartic acid protease (caspase) family. Sequential activation of caspases plays a central role in the execution-phase of cell apoptosis. Caspases exist as inactive proenzymes that undergo proteolytic processing at conserved aspartic residues to produce two subunits, large and small, that dimerize to form the active enzyme. It is the predominant caspase involved in the cleavage of amyloid-beta 4A precursor protein, which is associated with neuronal death in Alzheimer's disease. And the caspase-3 activation in heart failure sequentially cleaves SRF and generates a truncated SRF that appears to function as a dominant-negative transcription factor. Additionally, the caspase-3 influence on bone mineral density should be considered in any in vivo application of caspase-3 inhibitors to the treatment of human disease. In erythroid precursors undergoing terminal differentiation, Hsp70 prevents active CASP3 from cleaving GATA1 and inducing apoptosis.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
M00334-6 is guaranteed for Flow Cytometry, IHC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.1-0.5μg/ml
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml
Flow Cytometry (Fixed), 1-3μg/1x106 cells
Positive Control
WB: human placenta tissue,, human K562 whole cell,, human HepG2 whole cell,, human Raji whole cell
IHC: human tonsil tissue, human intestinal cancer tissue, human placenta tissue
FCM: HepG2 cell
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of Caspase-3 using anti-Caspase-3 antibody (M00334-6).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human placenta tissue lysates;
Lane 2: human K562 whole cell lysates;
Lane 3: human HepG2 whole cell lysates;
Lane 4: human Raji whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-Caspase-3 antigen affinity purified monoclonal antibody (Catalog # M00334-6) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for Caspase-3 at approximately 35KD. The expected band size for Caspase-3 is at 32KD.
Click image to see more details
Figure 2. IHC analysis of Caspase-3 using anti-Caspase-3 antibody (M00334-6).
Caspase-3 was detected in paraffin-embedded section of human tonsil tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-Caspase-3 Antibody (M00334-6) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 3. IHC analysis of Caspase-3 using anti-Caspase-3 antibody (M00334-6).
Caspase-3 was detected in paraffin-embedded section of human intestinal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-Caspase-3 Antibody (M00334-6) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 4. IHC analysis of Caspase-3 using anti-Caspase-3 antibody (M00334-6).
Caspase-3 was detected in paraffin-embedded section of human placenta tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-Caspase-3 Antibody (M00334-6) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 5. Flow Cytometry analysis of HepG2 cells using anti- Caspase-3 antibody (M00334-6).
Overlay histogram showing HepG2 cells stained with M00334-6 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with mouse anti- Caspase-3 Antibody (M00334-6, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Protein Target Info & Infographic
Gene/Protein Information For CASP3 (Source: Uniprot.org, NCBI)
Gene Name
CASP3
Full Name
Caspase-3
Weight
31.608kDa
Superfamily
peptidase C14A family
Alternative Names
Caspase-3; CASP-3; Apopain; Cysteine protease CPP32; CPP-32; Protein Yama; SREBP cleavage activity 1; SCA-1; CASP3; CPP32 CASP3 CPP32, CPP32B, SCA-1 caspase 3 caspase-3|CASP-3|CPP-32|PARP cleavage protease|SREBP cleavage activity 1|apopain|caspase 3, apoptosis-related cysteine peptidase|caspase 3, apoptosis-related cysteine protease|cysteine protease CPP32|procaspase3|protein Yama
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on CASP3, check out the CASP3 Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for CASP3: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-Caspase-3 Antibody Picoband® (monoclonal, 8B6) (M00334-6)
Hello CJ!
M00334-6 has been cited in 16 publications:
*The publications in this section are manually curated by our staff scientists. They may differ from Bioz's machine gathered results. Both are accurate. If you find a publication citing this product but is missing from this list, please let us know we will issue you a thank-you coupon.
Liu DX,Li PP,Guo JP,Li LL,Guo B,Jiao HB,Wu JH,Chen JM.Exosomes derived from HBV-associated liver cancer promote chemoresistance by upregulating chaperone-mediated autophagy.Oncol Lett.2019 Jan;17(1):323-331.doi:10.3892/ol.2018.9584.Epub 2018 Oct 16.PMID:30655770; PMCID: PMC6313222.
Species: Human
M00334-6 usage in article: APP:WB, SAMPLE:HEPG2 CELL, DILUTION:1:500
Zhang Z,Wang J,Zhu Y,Zhang H,Wang H.Astragaloside IV alleviates myocardial damage induced by type 2 diabetes via improving energy metabolism.Mol Med Rep.2019 Nov;20(5):4612-4622.doi:10.3892/mmr.2019.10716.Epub 2019 Oct 1.PMID:31702040; PMCID:PMC6797977.
Species: Human,Rat
M00334-6 usage in article: APP:WB, SAMPLE:MITOCHONDRIAL TISSUE AND H9C2 CELL, DILUTION:1:1000
Yu Y,Wang Y,Fei X,Song Z,Xie F,Yang F,Liu X,Xu Z,Wang G.All-Trans Retinoic Acid Prevented Vein Grafts Stenosis by Inhibiting Rb-E2F Mediated Cell Cycle Progression and KLF5-RARα Interaction in Human Vein Smooth Muscle Cells.Cardiovasc Drugs Ther.2020 Oct
Species: Human,New Zealand White Rabbit
M00334-6 usage in article: APP:WB, SAMPLE:HUVMSCS, DILUTION:1:1000
Liu L,Liu F,Sun Z,Peng Z,You T,Yu Z. LncRNA NEAT1 promotes apoptosis and inflammation in LPS-induced sepsis models by targeting miR-590-3p. Exp Ther Med.2020 Oct; 20(4):3290-3300. doi:10.3892/etm.2020.9079. Epub 2020 Jul 29.PMID:32855700;PMCID:PMC7444425.
Species: Human
M00334-6 usage in article: APP:WB, SAMPLE:H9C2 CELL, DILUTION:1:1000
Maternal separation exacerbates Alzheimer's disease-like behavioral and pathological changes in adult APPswe/PS1dE9 mice
MicroRNA-744 promotes prostate cancer progression through aberrantly activating Wnt/%u03B2-catenin signaling
In vitro and in vivo antitumor effects of acetylshikonin isolated from Arnebia euchroma (Royle) Johnst (Ruanzicao) cell suspension cultures
Enhanced anti-apoptosis and gut epithelium protection function of acidic fibroblast growth factor after cancelling of its mitogenic activity
Antenatal taurine reduces cerebral cell apoptosis in fetal rats with intrauterine growth restriction
An J, Liu T, She R, Wu Q, Tian J, Shi R, Hao W, Ren X, Yang Y, Lu Y, Yang Y, Wu Y. Oncotarget. 2017 Dec 17;9(4):4475-4484. doi: 10.18632/oncotarget.23381. eCollection 2018 Jan 12. Replication of hepatitis E virus in the ovary and promotion of oocy...
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Customer Reviews
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Customer Q&As
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16 Customer Q&As for Anti-Caspase-3 Antibody Picoband® (monoclonal, 8B6)
Question
See below the WB image, lot number and protocol we used for lymph using anti-Caspase-3 antibody (monoclonal, 8B6) M00334-6. Please let me know if you require anything else.
Verified Customer
Verified customer
Asked: 2020-02-19
Answer
Thank you very much for the data. Our lab team are working to resolve this as quickly as possible, and we appreciate your patience and understanding! You have provided everything we needed. Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2020-02-19
Question
Is there a BSA free version of anti-Caspase-3 antibody (monoclonal, 8B6) M00334-6 available?
Verified Customer
Verified customer
Asked: 2020-02-13
Answer
We appreciate your recent telephone inquiry. I can confirm that some lots of this anti-Caspase-3 antibody (monoclonal, 8B6) M00334-6 are BSA free. For now, these lots are available and we can make a BSA free formula for you free of charge. It will take 3 extra days to prepare. If you require this antibody BSA free again in future, please do not hesitate to contact me and I will be pleased to check which lots we have in stock that are BSA free.
Boster Scientific Support
Answered: 2020-02-13
Question
We purchased anti-Caspase-3 antibody (monoclonal, 8B6) for WB on tongue in the past. I am using human, and We intend to use the antibody for IHC-P next. My question regards examining tongue as well as lymph in our next experiment. Do you have any suggestion on which antibody would work the best for IHC-P?
Verified Customer
Verified customer
Asked: 2020-02-04
Answer
I took a look at the website and datasheets of our anti-Caspase-3 antibody (monoclonal, 8B6) and it seems that M00334-6 has been validated on human in both WB and IHC-P. Thus M00334-6 should work for your application. Our Boster satisfaction guarantee will cover this product for IHC-P in human even if the specific tissue type has not been validated. We do have a comprehensive range of products for IHC-P detection and you can check out our website bosterbio.com to find out more information about them.
Boster Scientific Support
Answered: 2020-02-04
Question
Is this M00334-6 anti-Caspase-3 antibody (monoclonal, 8B6) reactive to the isotypes of CASP3?
Verified Customer
Verified customer
Asked: 2019-12-23
Answer
The immunogen of M00334-6 anti-Caspase-3 antibody (monoclonal, 8B6) is E.coli-derived human Caspase-3 recombinant protein (Position: T67-D175). Human Caspase-3 shares 86% and 90% amino acid (aa) sequence identity with mouse and rat Caspase-3, respectively. Could you tell me which isotype you are interested in so I can help see if the immunogen is part of this isotype?
Boster Scientific Support
Answered: 2019-12-23
Question
We appreciate helping with my inquiry over the phone. Here are the WB image, lot number and protocol we used for lymph using anti-Caspase-3 antibody (monoclonal, 8B6) M00334-6. Let me know if you need anything else.
K. Miller
Verified customer
Asked: 2019-09-20
Answer
Thank you for the data. You have provided everything we needed. Our lab team are working to resolve your inquiry as quickly as possible, and we appreciate your patience and understanding! Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2019-09-20
Question
We are currently using anti-Caspase-3 antibody (monoclonal, 8B6) M00334-6 for human tissue, and we are satisfied with the WB results. The species of reactivity given in the datasheet says human. Is it true that the antibody can work on pig tissues as well?
Verified Customer
Verified customer
Asked: 2019-09-12
Answer
The anti-Caspase-3 antibody (monoclonal, 8B6) (M00334-6) has not been tested for cross reactivity specifically with pig tissues, but there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in pig you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2019-09-12
Question
How many vials do you suggest in IHC-P for 60 slides application using M00334-6 antibody?
Verified customer
Asked: 2019-07-24
Answer
The suggested working concentration for IHC-P is 1ug/ml. Our lab technicians usually add 50ul onto each slide. It takes 3ug of Anti-Caspase-3 Antibody Picoband (monoclonal, 8B6) M00334-6 for 60 slides.
Boster Scientific Support
Answered: 2019-07-26
Question
I see that the anti-Caspase-3 antibody (monoclonal, 8B6) M00334-6 works with IHC-P, what is the protocol used to produce the result images on the product page?
Verified Customer
Verified customer
Asked: 2019-06-21
Answer
You can find protocols for IHC-P on the "support/technical resources" section of our navigation menu. If you have any further questions, please send an email to [email protected]
Boster Scientific Support
Answered: 2019-06-21
Question
Is a blocking peptide available for product anti-Caspase-3 antibody (monoclonal, 8B6) (M00334-6)?
Verified Customer
Verified customer
Asked: 2019-05-28
Answer
We do provide the blocking peptide for product anti-Caspase-3 antibody (monoclonal, 8B6) (M00334-6). If you would like to place an order for it please contact [email protected] and make a special request.
Boster Scientific Support
Answered: 2019-05-28
Question
My question regarding product M00334-6, anti-Caspase-3 antibody (monoclonal, 8B6). I was wondering if it would be possible to conjugate this antibody with biotin. I would need it to be without BSA or sodium azide. I am planning on using a buffer exchange of sodium azide with PBS only. Would there be problems for me to conjugate the antibody and store it in -20 degrees in small aliquots?
Verified Customer
Verified customer
Asked: 2018-02-22
Answer
We suggest not storing this antibody with PBS buffer only in -20 degrees. If you want to store it in -20 degrees it is best to add some cryoprotectant like glycerol. If you want carrier free M00334-6 anti-Caspase-3 antibody (monoclonal, 8B6), we can provide it to you in a special formula with trehalose and/or glycerol. These molecules will not interfere with conjugation chemistry and provide a good level of protection for the antibody from degradation. Please be sure to specify this in your purchase order.
Boster Scientific Support
Answered: 2018-02-22
Question
I was wanting to use your anti-Caspase-3 antibody (monoclonal, 8B6) for IHC-P for human lymph on frozen tissues, but I want to know if it has been validated for this particular application. Has this antibody been validated and is this antibody a good choice for human lymph identification?
Verified Customer
Verified customer
Asked: 2017-12-27
Answer
It shows on the product datasheet, M00334-6 anti-Caspase-3 antibody (monoclonal, 8B6) has been tested for IHC-P, WB on human tissues. We have an innovator award program that if you test this antibody and show it works in human lymph in IHC-frozen, you can get your next antibody for free.
Boster Scientific Support
Answered: 2017-12-27
Question
My team were satisfied with the WB result of your anti-Caspase-3 antibody (monoclonal, 8B6). However we have been able to see positive staining in t-cell cytoplasm. using this antibody. Is that expected? Could you tell me where is CASP3 supposed to be expressed?
Verified Customer
Verified customer
Asked: 2017-12-14
Answer
According to literature, t-cell does express CASP3. Generally CASP3 expresses in cytoplasm. Regarding which tissues have CASP3 expression, here are a few articles citing expression in various tissues:
Cervix carcinoma, and Erythroleukemia, Pubmed ID: 23186163
Lymph, Pubmed ID: 15489334
T-cell, Pubmed ID: 7983002, 7774019
Tongue, Pubmed ID: 14702039
Boster Scientific Support
Answered: 2017-12-14
Question
Would M00334-6 anti-Caspase-3 antibody (monoclonal, 8B6) work on parafin embedded sections? If so, which fixation method do you recommend we use (PFA, paraformaldehyde, other)?
K. Edwards
Verified customer
Asked: 2016-09-05
Answer
As indicated on the product datasheet, M00334-6 anti-Caspase-3 antibody (monoclonal, 8B6) as been validated on IHC-P. It is best to use PFA for fixation because it has better tissue penetration ability. PFA needs to be prepared fresh before use. Long term stored PFA turns into formalin, as the PFA molecules congregate and become formalin.
Boster Scientific Support
Answered: 2016-09-05
Question
Would anti-Caspase-3 antibody (monoclonal, 8B6) M00334-6 work for IHC-P with lymph?
B. Rodriguez
Verified customer
Asked: 2016-03-30
Answer
According to the expression profile of lymph, CASP3 is highly expressed in lymph. So, it is likely that anti-Caspase-3 antibody (monoclonal, 8B6) M00334-6 will work for IHC-P with lymph.
Boster Scientific Support
Answered: 2016-03-30
Question
I am interested in to test anti-Caspase-3 antibody (monoclonal, 8B6) M00334-6 on human lymph for research purposes, then I may be interested in using anti-Caspase-3 antibody (monoclonal, 8B6) M00334-6 for diagnostic purposes as well. Is the antibody suitable for diagnostic purposes?
A. Brown
Verified customer
Asked: 2014-08-29
Answer
The products we sell, including anti-Caspase-3 antibody (monoclonal, 8B6) M00334-6, are only intended for research use. They would not be suitable for use in diagnostic work. If you have the means to develop a product into diagnostic use, and are interested in collaborating with us and develop our product into an IVD product, please contact us for more discussions.
Boster Scientific Support
Answered: 2014-08-29
Question
We have been able to see staining in human jejunal mucosa. Any tips? Is anti-Caspase-3 antibody (monoclonal, 8B6) supposed to stain jejunal mucosa positively?
C. Kulkarni
Verified customer
Asked: 2013-06-25
Answer
From literature jejunal mucosa does express CASP3. From Uniprot.org, CASP3 is expressed in jejunal mucosa, t-cell, tongue, lymph, cervix carcinoma erythroleukemia, among other tissues. Regarding which tissues have CASP3 expression, here are a few articles citing expression in various tissues:
Cervix carcinoma, and Erythroleukemia, Pubmed ID: 23186163
Lymph, Pubmed ID: 15489334
T-cell, Pubmed ID: 7983002, 7774019
Tongue, Pubmed ID: 14702039
Boster Scientific Support
Answered: 2013-06-25
