Caspase-3 Antibody

Immunohistochemistry-Paraffin: Caspase-3 Antibody [NB600-1235] - Formalin fixed paraffin embedded human tonsil stained with Caspase 3 antibody (NB600-1235).

Immunohistochemistry-Paraffin: Caspase-3 Antibody [NB600-1235] - Human tonsil.

• Reactivity: Hu, Mu, Rt, Po, Bv, Ca, Fi, Mk, Rb, Sh
• Applications: WB, ICC/IF, IHC, IHC-P
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1.0 mg/ml

Caspase-3 Antibody Summary

• Immunogen: A synthetic peptide corresponding to the cleavage site of human caspase 3 (amino acids 167-175), conjugated to KLH.
• Localization: Cytoplasmic and some nuclear.
• Specificity: This reacts with the active form of Caspase 3 (17 kDa protein).
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: CASP3
• Purity: Affinity purified

Applications/Dilutions

• Dilutions:
  • Western Blot
  • Immunocytochemistry/Immunofluorescence 1:10-1:500
  • Immunohistochemistry 1:10-1:500
  • Immunohistochemistry-Paraffin 1:50-1:100
• Application Notes: IHC (formalin/paraffin): staining of formalin-fixed tissues requires boiling tissue sections in 10mM citrate buffer, pH 6.0 (30 min at RT). Use in ICC/IF was reported in the scientific literature (PMID: 23813946). Use in Western blot reported in scientific literature (PMID: 29977195).
• Control:

  Jurkat Whole Cell Lysate
  Caspase-3 Overexpression Lysate
  Human Tonsil Whole Tissue Lysate (Adult Whole Normal)

Reactivity Notes

Fish reactivity reported in scientific literature (PMID: 28675853).

Packaging, Storage & Formulations

• Storage: Store at 4C. Do not freeze.
• Buffer: PBS (pH 7.4) and 0.2% BSA
• Preservative: 0.09% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Affinity purified

Alternate Names for Caspase-3 Antibody

• Apopain
• apoptosis-related cysteine protease
• CASP3
• CASP-3
• caspase 3, apoptosis-related cysteine peptidase
• Caspase3
• Caspase-3
• CPP32
• CPP-32
• CPP32B
• CPP32SREBP cleavage activity 1
• Cysteine protease CPP32
• EC 3.4.22
• EC 3.4.22.56
• LICE-1
• PARP cleavage protease
• procaspase3
• Protein Yama
• SCA-1
• YAMA

Background

Caspase 3 is a member of the interleukin-1 BETA-converting enzyme family. Caspase 3 is thought to be associated with induction of apoptosis.Caspase 3 is ubiquitously expressed and like other caspases is synthesized as an inactive (32kDa) proenzyme. Upon activation, Caspase 3 is cleaved at Asp28-Ser29 and Asp175-Ser176 thereby generating two subunits of 17kDa and 12kDa, respectively. Recent studies have implicated that Caspase 3 is associated with the induction of apoptosis. Activation of Caspase 3 occurs in response to variety of apoptotic inducers including Fas mediated apoptosis.

Limitations

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.

Publications for Caspase-3 Antibody (NB600-1235)(20)

Publications using NB600-1235

• Yaya M, Bird J, Idris B, Amer A. Nitric oxide mediated regulation of retinoic acid induced cartilage breakdown J Histol Histopathol Res Jul 10 2018 (IHC, Bovine)
• Onk D, Onk OA, Erol HS et al. Effect of melatonin on antioxidant capacity, inflammation and apoptotic cell death in lung tissue of diabetic rats Acta Cir Bras Apr 1 2018 [PMID: 29768540] (IHC-P, Rat)
• Ozcelik M, Erisir M, Guler O et al. The effect of curcumin on lipid peroxidation and selected antioxidants in irradiated rats Acta Vet. Brno Jan 16 2019 (IHC-P, Rat)
• COMAKLI S. GLUFOSINATE-BASED HERBICIDE TOXICITY CAUSE INCREASE CLEAVED CASPASE 3 IN RAT LIVER researchgate.net Jul 10 1905 (IHC, Rat)
• Reynolds JA, Henwood MK, Turtle JD et al. Brain-Dependent Processes Fuel Pain-Induced Hemorrhage After Spinal Cord Injury Front Syst Neurosci Sep 10 2019 [PMID: 31551720] (WB, Rat)
• Sevim, Comakli S, Taghizadehghalehjoughi A et al. An imazamox-based herbicide causes apoptotic changes in rat liver and pancreas. Toxicology Reports. Nov 1 2018 [PMID: 30560060] (IHC-P, Rat)
• Turtle JD, Strain MM, ReYnolds JA et al. Pain Input After Spinal Cord Injury (SCI) Undermines Long-Term Recovery and Engages Signal Pathways That Promote Cell Death. Front Syst Neurosci. Jun 21 2018 [PMID: 29977195] (WB, Rat)
• Tamma R, Ruggieri S, Annese T et al. Bcl6/p53 expression, macrophages/mast cells infiltration and microvascular density in invasive breast carcinoma Oncotarget Apr 27 2018 [PMID: 29854311] (IHC, Human)
• Altun S, ozdemir S, Arslan H. Histopathological effects, responses of oxidative stress, inflammation, apoptosis biomarkers and alteration of gene expressions related to apoptosis, oxidative stress, and reproductive system in chlorpyrifos-exposed common carp (Cyprinus carpio L.) Environ. Pollut. 2017 Jul 01 [PMID: 28675853] (ICC/IF, Fish)
• Aksu EH, Kandemir FM, Altun S et al. Ameliorative Effect of Carvacrol on Cisplatin-Induced Reproductive Damage in Male Rats. J. Biochem. Mol. Toxicol. 2016 Apr 25 [PMID: 27111473] (IHC-P, Rat)
  
  Details:
  Rabbit polyclonal active/cleaved Caspase-3 antibody was used at 1:400 dilution for IHC-P analysis of testes of Sprague-Dawley rats which were subjected to treatments with Cisplatin and/or Carvacrol/CARV, an antioxidant that is naturally found in some pla
• Aksu EH, Ozkaraca M, Kandemir FM et al. Mitigation of paracetamol-induced reproductive damage by chrysin in male rats via reducing oxidative stress Andrologia Feb 24 2016 [PMID: 26914515] (IHC-P, Rat)
• de Souza PC, Balasubramanian K, Njoku C et al. OKN-007 decreases tumor necrosis and tumor cell proliferation and increases apoptosis in a preclinical F98 rat glioma model J Magn Reson Imaging 2015 Apr 29 [PMID: 25920494] (IHC, Rat)
• Ozkaraca M, Ceribasi S, Ceribasi AO, Kilic A. The Role of Apoptosis and Autophagy in Bovine Abortions Associated with Brucella Spp. Acta Veterinaria-Beograd. 2016 (IHC-P, Bovine)
• Hunziker EB, Lippuner K, Keel MJ et al. A novel organ-slice culturing system to simulate meniscal repair: Proof of concept using a synovium-based pool of meniscoprogenitor cells. J. Orthop. Res. 2016 Jan 21 [PMID: 26790377] (IHC-P, Human)
• Kandemir FM, Ozkaraca M, Yildirim BA et al. Rutin attenuates gentamicin-induced renal damage by reducing oxidative stress, inflammation, apoptosis, and autophagy in rats. Ren Fail 2015 Apr [PMID: 25613739] (Rat)
• Buyuklu M, Kandemir F, Ozkaraca M et al. Benefical effects of lycopene against contrast medium-induced oxidative stress, inflammation, autophagy, and apoptosis in rat kidney. Hum Exp Toxicol. 2014 Sep 08 [PMID: 25205736]
• Buyuklu M, Mehmet Kandemir F, Ozkaraca M et al. Protective effect of curcumin against contrast induced nephropathy in rat kidney: what is happening to oxidative stress, inflammation, autophagy and apoptosis? Eur Rev Med Pharmacol Sci 3/10/2014 [PMID: 24610611] (IHC-P, Rat)
• Topcu I, Vatansever S, Bayram E et al. The effects of lornoxicam on neuroprotection following diffuse traumatic brain injury in rats. Turk Neurosurg 2013 [PMID: 24310460] (IHC-P, Rat)
• Wang L, Wu Y, Tan Y et al. Cytotoxic effects of the quinolone levofloxacin on rabbit meniscus cells. J Appl Toxicol 2013 Jul 1 [PMID: 23813946] (ICC/IF, Rabbit)
• Jee D, Lee WK. Inhibitory Effect of intravitreal Injection of Bevacizumab on Nerve Growth Factor. Current Eye Research. 2011 Oct 31. [PMID: 22040304]

FAQs for Caspase-3 Antibody (NB600-1235). (Showing 1 - 4 of 4 FAQs).

1. Any reason for cleaved caspase 3 to be seen as purely nuclear staining or is this just an artifact?
   • Caspase 3 can translocate from the cytoplasm to the nucleus, usually during apoptosis. We do offer a 100% guarantee that this antibody will be useful as stated on the datasheet also.
2. The datasheet says that it reacts with the active form of Caspase 3 (17 kDa protein), but every time when I do the WB, I get two bands between 15kDa and 20kDa. The different company's cleaved caspase 3 antibody reacts with 17 and 19 kDa protein, so I wonder which band is specific to yours.
   • Regarding your question, NB600-1235 has been exclusively validated for IHC, not western blot and it is supposed to detect the active form of caspase3. The inactive Caspase 3 is a 31 kD protein, a heterotetramer that consists of two anti-parallel arranged heterodimers, each one formed by a 17 kDa (p17) and a 12 kDa (p12). When it is Cleaved by granzyme B, it generates the two active subunits. As a result the two bands you are observing on the gel should be correct. However, we haven't tested this antibody for western blot.
3. I'd like to get the information about the active/cleaved Caspase 3 Antibody (NB600-1235). The datasheet says that it reacts with the active form of Caspase 3 (17 kDa protein), but every time when I do the WB, I get two bands between 15kDa and 20kDa. The different company's cleaved caspase 3 antibody reacts with 17 and 19 kDa protein, so I wonder which band is specific to yours?
   • According to the data I could find for the active/cleaved caspase 3, the active form should appear at approximately 17kDa and also has a smaller subunit seen at approximately 12kDa. The inactive form is approximately 32kDa. These are the theoretical weights and can vary based on type of gel and samples they are run on. In most cases, it looks like most antibodies detect the active forms at 19 and 17kDa respectively. With this particular product, NB600-1235, we have not yet validated it for use in western blot, so we unfortunately do not have any information for the location of bands.
4. This is a Rabbit antibody right so Do you think I can use it on rabbit tissue?
   • This antibody does react to rabbit but if you are using it with an anti-rabbit secondary, you will almost certainly see some background staining. I know that mouse-on-mouse blocking systems exist, but I am not sure if there is a rabbit equivalent at this time. The best way to use NB600-1235 on rabbit tissues would be to have it directly conjugated to biotin, HRP, or whatever detection method you are planning to use. This will eliminate the need for an anti-rabbit secondary and should give you little to no background. We do offer a conjugation service, so if you would like to have this antibody conjugated, please let me know what you would like to have this antibody conjugated to and I can get you a quote, email bulk@novusbio.com.

Bioinformatics

• Gene Symbol: CASP3
• Entrez:
  • 836(Human)
  • 12367(Mouse)
  • 25402(Rat)
• Uniprot:
  • P42574 ()
  • P42574 ()