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Caspase-3 Antibody (31A1067) - (Pro and Active) - BSA Free

Novus Biologicals, part of Bio-Techne | Catalog # NB100-56708

Novus Biologicals, part of Bio-Techne
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NB100-56708
NB100-56708SS

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat, Porcine, Chicken, Chinese Hamster, Mammal

Cited:

Human, Mouse, Rat, Avian - Chicken, Hamster - Cricetulus (Chinese Hamster), Mammal, Rabbit

Applications

Validated:

CyTOF-ready, Electron Microscopy, Flow Cytometry, Hematoxylin and Eosin Stain, Immunoblotting, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry Free-Floating, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Knockdown Validated, Knockout Validated, Simple Western, Western Blot

Cited:

Block/Neutralize, Electron Microscopy, Flow Cytometry, Hematoxylin and Eosin Stain, IF/IHC, Immunoblotting, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry Free-Floating, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Knockdown Validated, Simple Western, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # 31A1067

Format

BSA Free

Concentration

1 mg/ml

Product Specifications

Immunogen

This Caspase-3 Antibody (31A1067) - (Pro and Active) was developed against full-length recombinant human caspase-3 protein.

Reactivity Notes

Chicken reactivity reported in scientific literature (PMID: 30298003).

Specificity

The antibody detects both pro Caspase-3 (~32 kDa) and the large subunit of the active/cleaved form (~14-21 kDa) of Caspase-3.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

Theoretical MW

31.7 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for Caspase-3 Antibody (31A1067) - (Pro and Active) - BSA Free

Western Blot Detection of Pro and Active Caspase-3 in Whole Cell Protein From Treated Jurkat Cells

Western Blot Detection of Pro and Active Caspase-3 in Whole Cell Protein From Treated Jurkat Cells

Image of Caspase-3 Antibody (31A1067) - (Pro and Active). Whole cell protein from Jurkat cells treated with and without 2 uM staurosporine as indicated was separated on a 4-15% gel by SDS-PAGE, transferred to 0.2 um PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 5 ug/ml anti-Caspase 3 in 1% milk, and detected with an anti-mouse HRP secondary antibody using a Femto sensitivity chemiluminescence reagent. Note the detection of both pro-caspase 3 at 35 kDa and the cleaved active caspase 3 at 15-17 kDa.
Immunohistochemical Staining of Caspase-3 in Paraffin Embedded Human Spleen

Immunohistochemical Staining of Caspase-3 in Paraffin Embedded Human Spleen

Tissue section of human spleen using 1:200 dilution of Caspase-3 antibody (clone 31A1067). The staining was developed with HRP labeled anti-mouse IgG secondary antibody and DAB reagent, and nuclei of cells were counter-stained with hematoxylin. This Caspase 3 antibody generated primarily a specific cytoplasmic staining in a subset of spleenocytes with some nuclear signal in a few cells.
Detection of Caspase-3 in HeLa Cell Lysate by Simple Western

Detection of Caspase-3 in HeLa Cell Lysate by Simple Western

Simple Western lane view shows a specific band for Caspase-3 Antibody (31A1067) - (Pro and Active) in 0.1 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230kDa separation system.

Applications for Caspase-3 Antibody (31A1067) - (Pro and Active) - BSA Free

Application
Recommended Usage

Electron Microscopy

reported in scientific literature (PMID 27450722)

Flow Cytometry

reported in scientific literature (PMID 27429862)

Hematoxylin and Eosin Stain

reported in scientific literature (PMID 28186963)

Immunoblotting

reported in scientific literature (PMID 28500555)

Immunocytochemistry/ Immunofluorescence

reported in scientific literature (PMID 23840553)

Immunohistochemistry

1:10 - 1:500

Immunohistochemistry Free-Floating

reported in scientific literature (PMID 31771656)

Immunohistochemistry-Frozen

1:10 - 1:500. Use reported by customer review

Immunohistochemistry-Paraffin

1:10 - 1:500. Use reported in scientific literature (PMID 28500555)

Knockdown Validated

reported in scientific literature (PMID 32867814)

Simple Western

1:50

Western Blot

1 - 5 ug/ml
Application Notes
The large subunit of the cleaved form may appear as one or two or even as a stack of bands depending on the presence or absence of the Caspase-3 pro-domain. It is highly recommended that a maximum sensitivity ECL substrate (Femto sensitive) be used for efficient detection of this antibody in Western blot applications. In Simple Western only 10 - 15 ul of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue. This antibody is CyTOF ready.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 3 reviews rated 4.7 using NB100-56708 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

BSA Free

Preservative

0.05% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: Caspase-3

CPP32 encodes a member of the cysteine-aspartic acid protease (caspase) family that plays a key role in apoptosis. Caspase 3 (also known as CPP32, Apopain, Yama, and PARP cleavage protease) is the most extensively studied apoptotic protein among caspase family members (1-2). Caspase 3 is synthesized as an inactive proenzyme that is processed in cells undergoing apoptosis by self-proteolysis and/or cleavage by other upstream proteases (e.g. Caspases 8, 9 and 10). Alternative splicing of CPP32 results in two transcript variants which encode the same protein. The processed form of Caspase 3 consists of large (theoretical molecular weight 17kD) and small (theoretical molecular weight 12kD) subunits which associate to form an active enzyme. Caspase 3 is cleaved at Asp28/Ser29 and Asp175/Ser176. The active Caspase 3 proteolytically cleaves and activates other caspases (e.g. Caspases 6, 7 and 9), as well as relevant targets in the cells (e.g. PARP and DFF). Variations in levels of Caspase 3 have been reported in cells of short-lived nature and those with a longer life cycle. Caspase 3 is the predominant caspase involved in the cleavage of amyloid beta 4A precursor protein, which is associated with neuronal death in Alzheimer's disease (3).

References

1.Mu, N., Lei, Y., Wang, Y., Wang, Y., Duan, Q., Ma, G., . . . Su, L. (2019). Inhibition of SIRT1/2 upregulates HSPA5 acetylation and induces pro-survival autophagy via ATF4-DDIT4-mTORC1 axis in human lung cancer cells. Apoptosis, 24(9-10), 798-811. doi:10.1007/s10495-019-01559-3

2.Sun, C. M., Enkhjargal, B., Reis, C., Zhou, K. R., Xie, Z. Y., Wu, L. Y., . . . Zhang, J. H. (2019). Osteopontin attenuates early brain injury through regulating autophagy-apoptosis interaction after subarachnoid hemorrhage in rats. CNS Neurosci Ther, 25(10), 1162-1172. doi:10.1111/cns.13199

3.Louneva, N., Cohen, J. W., Han, L. Y., Talbot, K., Wilson, R. S., Bennett, D. A., . . . Arnold, S. E. (2008). Caspase-3 is enriched in postsynaptic densities and increased in Alzheimer's disease. Am J Pathol, 173(5), 1488-1495. doi:10.2353/ajpath.2008.080434

Alternate Names

Apopain, CASP3, Caspase3, CPP32, LICE-1, YAMA

Gene Symbol

CASP3

Additional Caspase-3 Products

Product Documents for Caspase-3 Antibody (31A1067) - (Pro and Active) - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for Caspase-3 Antibody (31A1067) - (Pro and Active) - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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