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Caspase-3 Antibody (CPP32 4-1-18) - BSA Free

Catalog # NB500-210 | Novus Biologicals a Bio-Techne Brand

Key Product Details

Validated by

Biological Validation

Species Reactivity

Human, Mouse, Rat

Applications

Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Immunoprecipitation, Simple Western, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2a Kappa Clone # CPP32 4-1-18

Format

BSA Free

Concentration

1.0 mg/ml

Product Summary for Caspase-3 Antibody (CPP32 4-1-18) - BSA Free

Immunogen

This Caspase-3 Antibody (CPP32 4-1-18) was developed against full-length recombinant human Caspase 3 [UniProt# P42574].

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2a 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 (CPP32 4-1-18) - BSA Free

Western Blot: Caspase-3 Antibody (CPP32 4-1-18) [NB500-210] - ALM induces HIF-1alpha-dependent apoptosis in PC3 cells. PC3 cells were treated with ALM for 24 h and 48 h at 0.1 uM under normoxia and hypoxia, followed by Western blot for caspase-3. Image collected and cropped by CiteAb from the following publication (https://www.mdpi.com/2073-4409/8/5/439), licensed under a CC-BY license.
Simple Western: Caspase-3 Antibody (CPP32 4-1-18) [NB500-210] - Lane view shows a specific band for Caspase 3 in 0.5 mg/ml of Hek293 lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.
Immunohistochemistry-Paraffin: Caspase-3 Antibody (CPP32 4-1-18) [NB500-210] - Analysis of a FFPE human spleen section using 1:200 dilution of . The staining was developed using HRP conjugated anti-mouse secondary antibody and DAB reagent. This Caspase 3 antibody generated a specific staining in the cytoplasm of various spleenocytes.

Applications for Caspase-3 Antibody (CPP32 4-1-18) - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

2 ug/ml

Immunohistochemistry

1:200-1:500

Immunohistochemistry-Frozen

reported in scientific literature (PMID 31242448)

Immunohistochemistry-Paraffin

1:10-1:500

Immunoprecipitation

2 ug / mg lysate

Simple Western

1:2000

Western Blot

1:500-1:1000
Application Notes
A band is seen at ~32 kDa for the inactive form of Caspase 3 and ~17-22 kDa for the active form of Caspase 3 in Western Blot. (See protocol for additional information).

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 4 reviews rated 4 using NB500-210 in the following applications:

Published Applications

Read 24 publications using NB500-210 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

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

Entrez Gene IDs

836 (Human); 12367 (Mouse); 25402 (Rat)

Gene Symbol

CASP3

UniProt

Product Documents for Caspase-3 Antibody (CPP32 4-1-18) - 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 (CPP32 4-1-18) - 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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Protocols

View specific protocols for Caspase-3 Antibody (CPP32 4-1-18) - BSA Free (NB500-210):

[[URL:https://www.novusbio.com/products/caspase-3-antibody-cpp32-4-1-18_nb500… Antibody ]]
Procedure Guide for NB 500-210 Monoclonal anti-Caspase 3

Western Protocol

1) Run a 10-*15% SDS polyacrylamide gel, loading ~20 ug of cell extract per lane.
2) Transfer the proteins to a membrane.
3) Block the membrane in PT-T20 [20 mM Tris-HCl, pH 7.4 / 150 mM NaCl/0.5% Tween 20] + 5% NFDM [non-fat dry milk], for 3 hour at room temperature (RT), shaking gently.
4) Rinse the membrane twice with PT-T20.
5) Incubate the membrane in anti-Caspase 3 [NB 500-210], diluted 1:500-1:1,000 in PT-T20 + 5% NFDM, for 60 minutes at RT.
6) Wash the membrane for 15 minutes, 3 times, in PT-T20 at RT.
7) Incubate the membrane in anti-mouse conjugated to HRP (secondary antibody), diluted in PT-T20 + 5% NFDM for 60 minutes at RT.
8) Wash the membrane for 15 minutes, 3 times, in PT-T20 at RT.
9) Develop membrane in chemiluminescent reagents, as instructed by kit-vendor.

*Positive control(s): Human kidney 293 cells

To Activate the Cell Extracts:
Bring the extract to a final concentration of 5 mM dATP.
Incubate the extracts at 37C for 15-30 minutes.

FAQs for Caspase-3 Antibody (CPP32 4-1-18) - BSA Free

Showing  1 - 2 of 2 FAQs Showing All
  • Q: I am looking for a mouse antibody against active caspase 3. This antibody you have is one of the few mouse anti-caspase 3 antibodies that I can find, but the information about it does not specify whether it is against the active form of the protein. I must have an antibody against active caspase 3, can you tell me whether this is the case for your antibody (NB500-210).

    A: I have pulled up information about this antibody and it appears that it will detect both the active and non-active forms of the protein. I don't know if you are looking for one that is only specific to the active and will not detect the inactive, or if as long as it detects the active it will work.

  • Q: Which is the best retrieval buffer for this antibody to IHC? In addition, which is the dilution for it?

    A: Antigen retrieval should be performed using citrate buffer at a pH of 6.0. Our protocol for this can be found here: Antigen Retrieval Protocol.

Showing  1 - 2 of 2 FAQs Showing All
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