Anti-Caspase-3 Antibody, clone 3J16 , ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1221 - recombinant, expressed in HEK 293 cells
Synonym: Apopain; CASP-3; CPP-32; Cysteine protease CPP32; EC:3.4.22.56; Protein Yama; SCA-1; SREBP cleavage activity 1
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human tonsil (A) and Negative control (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1221, Anti-Caspase-3, clone 3J16 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in subsets of cells within germinal center and epithelium of human tonsil tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human tonsil (A) and Negative control (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1221, Anti-Caspase-3, clone 3J16 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in subsets of cells within germinal center and epithelium of human tonsil tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human tonsil (A) and Negative control (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1221, Anti-Caspase-3, clone 3J16 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in subsets of cells within germinal center and epithelium of human tonsil tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) mouse kidney (A) and Negative control (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1221, Anti-Caspase-3, clone 3J16 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic / membranous staining was observed in subsets of tubules of mouse kidney tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) mouse kidney (A) and Negative control (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1221, Anti-Caspase-3, clone 3J16 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic / membranous staining was observed in subsets of tubules of mouse kidney tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) mouse kidney (A) and Negative control (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1221, Anti-Caspase-3, clone 3J16 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic / membranous staining was observed in subsets of tubules of mouse kidney tissue sections.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of Jurkat cells was performed using a 1:100 dilution of Cat. No. ZRB1221, Anti-Caspase-3, clone 3J16 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus and cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of Jurkat cells was performed using a 1:100 dilution of Cat. No. ZRB1221, Anti-Caspase-3, clone 3J16 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus and cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of Jurkat cells was performed using a 1:100 dilution of Cat. No. ZRB1221, Anti-Caspase-3, clone 3J16 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus and cytoplasm.
Western Blotting Enhanced Validation - Recombinant Antibody Technology Lysate from Jurkat cells treated with Camptothecin (1 μM for 3 h) was probed with Cat. No. ZRB1221, Anti-Caspase-3, clone 3J16 ZooMAb® Rabbit Monoclonal (1:1,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrows indicate Caspase-3 (~31 kDa; 17 kDa p17 subunit).
Flow Cytometry Enhanced Validation - Recombinant Antibody Technology Staining of one million Jurkat cells was performed using 0.1 μg of Cat. No. ZRB1221, Anti-Caspase-3, clone 3J16 ZooMAb® Rabbit Monoclonal (Yellow histogram) or the equivalent amount of a Rabbit IgG isotype control (Grey histogram), followed by a PE-conjugated Donkey Anti-Rabbit secondary antibody.
12 Principles of Green Chemistry: Principle 1—Waste Prevention. This product has a smaller environmental footprint through reduction of waste in the manufacturing process.
12 Principles of Green Chemistry: Principle 4—Designing Safer Chemicals
Chemical products should be designed to affect their desired function while minimizing their toxicity.
Chemical products should be designed to affect their desired function while minimizing their toxicity.
12 Principles of Green Chemistry: Principle 6—Design for Energy Efficiency. This product was re-engineered to utilize less energy in the manufacturing process.
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| biological source | rabbit (recombinant) |
| clone | 3J16, recombinant monoclonal |
| conjugate | unconjugated |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| form | lyophilized |
| greener alternative category | Aligned |
| greener alternative product characteristics | Waste Prevention Designing Safer Chemicals Design for Energy Efficiency Learn more about the Principles of Green Chemistry . |
| isotype | IgG |
| mol wt | calculated mol wt 31.61 kDa |
| observed mol wt ~31 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human, rat |
| species reactivity (predicted by homology) | mouse, bovine |
| storage temp. | 2-8°C |
| technique(s) | flow cytometry: suitable |
| immunocytochemistry: suitable | |
| immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable | |
| western blot: suitable | |
| UniProt accession no. | P42574  |
| Application: | Anti-Caspase-3, clone 3J16 ZooMAb, Cat. No. ZRB1221, is a highly specific rabbit recombinant monoclonal antibody that targets Caspase-3 and has been tested for use in Flow Cytometry, Immunocytochemistry, Immunohistochemistry (Paraffin), and Western Blotting. |
| Application: | Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected Caspase-3 in human tonsil and human kidney tissue sections. Flow Cytometry Analysis: 0.1 μg from a representative lot detected Caspase-3 in Jurkat cells. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected Caspase-3 in Jurkat cells. Note: Actual optimal working dilutions must be determined by end user as specimens and experimental conditions may vary. |
| Disclaimer: | Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals. |
| General description: | We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information. |
| General description: | ZooMAb antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb antibodies are reliably available and ready to ship when you need them. Learn more about ZooMAb here. |
| Immunogen: | KLH-conjugated linear peptide corresponding to 12 amino acids from the p17 subunit of human Caspase-3. |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS with 5% Trehalose, normal appearance a coarse or translucent resin. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 μL. |
| Reconstitution: | 300 μg/mL after reconstitution at 25 μL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type. |
| Specificity: | Clone 3J16 is a ZooMAb rabbit recombinant monoclonal antibody that specifically detects Caspase-3. It targets an epitope within 12 amino acids from the p17 subunit. |
| Storage and Stability: | Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 μL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws. |
| Target description: | Caspase-3 (UniProt: P42574; also known as EC:3.4.22.56, CASP-3, Apopain, Cysteine protease CPP32, CPP-32, Protein Yama, SREBP cleavage activity 1, SCA-1) is encoded by the CASP3 (also known as CPP32) gene (Gene ID: 836) in human. Cysteine-aspartic proteases or Caspases play essential roles in apoptosis, necrosis, and inflammation. Historically, caspases were numbered in the order in which they were identified. Caspase-3 is a heterotetrameric enzyme that consists of two anti-parallel arranged heterodimers, each one formed by a 17 kDa (p17) and a 12 kDa (p12) subunit. Caspase-3 is initially produced with a propeptide sequence (aa 1-9), the removal of which yields the 268 amino acids caspase-3 proenzyme. Upon activation, the proenzyme is proteolytically cleaved first between Asp175-Ser176 to generate a p20 (aa 10-175) fragment and the p12 (aa 176-277) subunit. Further cleavage of the p20 fragment between Asp28-Ser29 produces the p17 (aa 29-175) subunit. The p17 and p12 subunits dimerize and forms the active caspase-3 enzyme. Caspase-3 has a strict requirement for an Asp residue at positions P1 and P4. It has a preferred cleavage sequence of Asp-Xaa-Xaa-Asp-|- with a hydrophobic amino-acid residue at P2 and a hydrophilic amino-acid residue at P3, although Val or Ala are also accepted at this position. Caspase-3 is involved in the activation cascade of caspases responsible for apoptosis execution. At the onset of apoptosis it proteolytically cleaves poly(ADP-ribose) polymerase (PARP) at a Asp216-|-Gly217 bond. Caspase-3 mediates the proteolytic activation of caspases-6 and -7, while caspase-3 itself is processed and activated by caspase-8, -9, or -10. It is the predominant caspase involved in the cleavage of amyloid-beta 4A precursor protein that is associated with neuronal death in Alzheimer′s disease. This ZooMAb recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. |
| WGK Germany | WGK 1 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |