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

Novus Biologicals | Catalog # NBP2-33244

Novus Biologicals
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Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

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

Cited:

Human, Mouse

Applications

Knockout Validated, Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Simple Western, Knockdown Validated

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # 31A1067

Format

Azide and BSA Free
View all Caspase-3 Primary Antibodies »

Product Specifications

Immunogen

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

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) - Azide and BSA Free

Western Blot Analysis of Caspase-3 in TSGH 8301 Cells After Multiple Treatment Types

Western Blot Analysis of Caspase-3 in TSGH 8301 Cells After Multiple Treatment Types

Paclitaxel in combination with MWE retarded tumor growth in a human bladder carcinoma TSGH 8301 xenograft model. The levels of total (t-PTEN) and phospho-PTEN (p-PTEN) and Caspase 3 in the tumor specimens were determined by Western blotting and then quantified using beta-actin as the protein loading control; the results are expressed as a percentage of the control. Image collected and cropped by CiteAb from the following publication (https://www.nature.com/articles/srep20417), licensed under a CC-BY license. Image using the standard format of this product.
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. Image using the standard format of this product.
Flow Cytometry of HeLa Cells Stained with Caspase-3 Antibody

Flow Cytometry of HeLa Cells Stained with Caspase-3 Antibody

An intracellular stain was performed on HeLa cells with Caspase-3 Antibody (31A1067) - (Pro and Active) NB100-56708 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature, followed by mouse F(ab)2 IgG (H+L) APC-conjugated secondary antibody (F0101B, R&D Systems). Image using the standard format of this product.
Detection of Caspase-3 in Multiple Human Tissues by Western Blot

Detection of Caspase-3 in Multiple Human Tissues by Western Blot

Western blot of Caspase-3 in multiple human tissues. The tissues shown are A) brain, B) heart, C) intestine, D) kidney, E) liver, F) lung, G) muscle, H) stomach, I) spleen, J) ovary, and K) testis.
Western Blot Detection of Caspase-3 in Human, Mouse and Rat Heart Lysate

Western Blot Detection of Caspase-3 in Human, Mouse and Rat Heart Lysate

Lanes 1, 2 and 3 demonstrate the species cross-reactivity of the antibody in human, mouse and rat heart lysate, respectively.
Western Blot Detection of Caspase-3 in HeLa Cells Treated with Staurosporine

Western Blot Detection of Caspase-3 in HeLa Cells Treated with Staurosporine

Analysis for detection of Caspase-3 activation in HeLa cells. Cells were treated with 2mM staurosporine for different time periods. Caspase-3 activation is determined by cleavage of procaspase-3, which generates 17 and 12kDa, larger and smaller catalytic subunit, respectively.
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. Image using the standard format of this product.
Western Blotting of Caspase-3 in VP-16 Treated and Untreated Jurkat Cells

Western Blotting of Caspase-3 in VP-16 Treated and Untreated Jurkat Cells

Lysates of Jurkat human acute T cell leukemia cell line untreated (-) or treated (+) with VP-16. PVDF membrane was probed with 0.1 ug/mL of mouse monoclonal Caspase-3 Antibody (31A1067) - (Pro and Active) (NB100-56708, Novus Biologicals) followed by 1:2000 dilution donkey anti-mouse IgG. Image using the standard format of this product.
Immunohistochemical Analysis of Caspase-3 in Paraffin Embedded Human Breast Cancer

Immunohistochemical Analysis of Caspase-3 in Paraffin Embedded Human Breast Cancer

Analysis in human breast cancer stained at 4ug/ml. Localization can be cytoplasmic and nuclear. Staining in the nucleus is considered to be an indication of active Caspase-3. In most cell types and model systems, cells with active Caspase-3 are undergoing apoptosis.
Immunohistochemical Staining of Caspase-3 in Paraffin Embedded Normal Colon

Immunohistochemical Staining of Caspase-3 in Paraffin Embedded Normal Colon

Analysis in normal colon stained 4ug/ml. Localization can be cytoplasmic and nuclear. Staining in the nucleus is considered to be an indication of active Caspase-3. In most cell types and model systems, cells with active Caspase-3 are undergoing apoptosis.
Immunohistochemical Detection of Caspase-3 in Paraffin Embedded Human Thymus

Immunohistochemical Detection of Caspase-3 in Paraffin Embedded Human Thymus

Analysis in the human thymus after heat-induced antigen retrieval at 5 ug/ml.
Immunohistochemical Analysis of Caspase-3 in Paraffin Embedded Human Bladder

Immunohistochemical Analysis of Caspase-3 in Paraffin Embedded Human Bladder

Caspase-3 was detected in immersion fixed paraffin-embedded sections of human bladder tissue using 1:50 dilution of mouse aCaspase-3 Antibody (31A1067) - (Pro and Active) (NB100-56708), for 1 hour at room temperature followed by anti-mouse IgG VisUCyte HRP polymer(VC001). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Image using the standard format of this product.
Flow Cytometry of NIH3T3 Cells Stained with Alexa Fluor 700 Conjugated Caspase-3 Antibody

Flow Cytometry of NIH3T3 Cells Stained with Alexa Fluor 700 Conjugated Caspase-3 Antibody

An intracellular stain was performed on NIH3T3 cells with Caspase-3 Antibody (31A1067) - (Pro and Active) Antibody NB100-56708AF700 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 10 ug/mL for 30 minutes at room temperature. Both antibodies were directly conjugated to Alexa Fluor 700.
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. Image using the standard format of this product.

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

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

reported in scientific literature (PMID 23840553)

Immunohistochemistry

1:10-1:500

Immunohistochemistry-Frozen

1:10-1:500. Use reported in scientific literature (Zhang et al)

Immunohistochemistry-Paraffin

1:10-1:500. Use reported in scientific literature (Lee et al)

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.

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS - BSA free

Format

Azide and BSA Free

Preservative

No Preservative

Concentration

1.0 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) - Azide and BSA Free

Certificate of Analysis

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Product Specific Notices for Caspase-3 Antibody (31A1067) - (Pro and Active) - Azide and 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.

Related Research Areas

Cysteine Proteases and Regulators

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

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