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

Novus Biologicals | Catalog # NB500-210

Novus Biologicals

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse

Applications

Validated:

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

Cited:

Immunohistochemistry-Paraffin, Western Blot, IF/IHC, IHC-F

Label

Unconjugated

Antibody Source

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

Format

BSA Free
View all Caspase-3 Primary Antibodies »

Product Specifications

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.

Description

Novus Biologicals Mouse Caspase-3 Antibody (CPP32 4-1-18) - BSA Free (NB500-210) is a monoclonal antibody validated for use in IHC, WB, ICC/IF, Simple Western and IP. Anti-Caspase-3 Antibody: Cited in 23 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.

Scientific Data Images for Caspase-3 Antibody (CPP32 4-1-18) - BSA Free

Western Blot: Caspase-3 Antibody (CPP32 4-1-18)BSA Free [NB500-210]

Western Blot: Caspase-3 Antibody (CPP32 4-1-18)BSA Free [NB500-210]

Caspase-3-Antibody-CPP32-4-1-18-Western-Blot-NB500-210-img0008.jpg
Simple Western: Caspase-3 Antibody (CPP32 4-1-18)BSA Free [NB500-210]

Simple Western: Caspase-3 Antibody (CPP32 4-1-18)BSA Free [NB500-210]

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) - BSA Free [NB500-210]

Immunohistochemistry-Paraffin: Caspase-3 Antibody (CPP32 4-1-18) - BSA Free [NB500-210]

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.
Immunohistochemistry-Paraffin: Caspase-3 Antibody (CPP32 4-1-18) - BSA Free [NB500-210]

Immunohistochemistry-Paraffin: Caspase-3 Antibody (CPP32 4-1-18) - BSA Free [NB500-210]

Immunohistochemistry-Paraffin: Caspase-3 Antibody (CPP32 4-1-18) [NB500-210] - Caspase-3 was detected in immersion fixed paraffin-embedded sections of human bladder 1:300 dilution of mouse monoclonal Caspase-3 Antibody (CPP32 4-1-18) (NB500-210, Novus Biologicals), 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).
Immunohistochemistry-Frozen: Caspase-3 Antibody (CPP32 4-1-18) - BSA Free [NB500-210]

Immunohistochemistry-Frozen: Caspase-3 Antibody (CPP32 4-1-18) - BSA Free [NB500-210]

Immunohistochemistry-Frozen: Caspase-3 Antibody (CPP32 4-1-18) - BSA Free [NB500-210] - Analysis of murine spleen with Caspase-3 (green), CD3 (red) and DAPI (blue). Image from verified customer review.
Western Blot: Caspase-3 Antibody (CPP32 4-1-18)BSA Free [NB500-210]

Western Blot: Caspase-3 Antibody (CPP32 4-1-18)BSA Free [NB500-210]

Western Blot: Caspase-3 Antibody (CPP32 4-1-18) [NB500-210] - Detection of Caspase (19 and 35 kDa) from HEK293 cell extract using (NB500-210). Lanes 1 and 2 contain inactive and active Caspase, respectively.
Immunohistochemistry-Paraffin: Caspase-3 Antibody (CPP32 4-1-18) - BSA Free [NB500-210]

Immunohistochemistry-Paraffin: Caspase-3 Antibody (CPP32 4-1-18) - BSA Free [NB500-210]

Immunohistochemistry-Paraffin: Caspase-3 Antibody (CPP32 4-1-18) [NB500-210] - Rat epithelial cells of the tongue base. Antigen retrieval method: Citrate buffer.

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.
See Simple Western Antibody Database for Simple Western validation: Tested in Hek293 lysate 0.5 mg/mL, separated by Size, antibody dilution of 1:2000, apparent MW was 40 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.

Reviewed Applications

Read 4 reviews rated 4 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

P42574 (Mouse)

Additional Caspase-3 Products

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 or batch number in the search box below.

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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.

Related Research Areas

Cysteine Proteases and Regulators

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

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

4 out of 5
4 Customer Ratings
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Customer Images

Showing  1 - 4 of 4 reviews Showing All
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  • Caspase-3 Antibody (CPP32 4-1-18) - BSA Free
    Name: Anonymous
    Application: Immunohistochemistry-Frozen
    Sample Tested: Mouse Spleen
    Species: Mouse
    Verified Customer | Posted 12/05/2022
    Murine spleen with Caspase 3 (green), CD3 (red) and DAPI (blue)
    Caspase-3 Antibody (CPP32 4-1-18) - BSA Free NB500-210
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: Human cancer cell whole cell lysate
    Species: Human
    Verified Customer | Posted 08/23/2015
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: Human Hepatoblastoma
    Species: Human
    Verified Customer | Posted 12/21/2011
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: Human
    Species: Human
    Verified Customer | Posted 11/21/2011

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Protocols

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


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.

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

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

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    • 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.
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