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Human/Mouse Active Caspase-3 Antibody

R&D Systems, part of Bio-Techne | Catalog # AF835

R&D Systems, part of Bio-Techne
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AF835
AF835-SP

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse, Rat, Porcine, Avian - Chicken, Canine, Chicken, Equine, Feline, Fish, Fish - Cyprinus carpio (Carp), Fish - Danio rerio (Zebrafish), Fish - Oncorhynchus mykiss (Rainbow Trout), Guinea Pig, Ovine, Primate, Transgenic Mouse

Applications

Validated:

Immunocytochemistry, Immunohistochemistry

Cited:

Flow Cytometry, Functional Assay, Immunocytochemistry, Immunocytochemistry/ Immunofluorescence, Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Immunoprecipitation, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Product Specifications

Immunogen

KLH-coupled human Caspase-3 synthetic peptide
CRGTELDCGIETD
Accession # P42574

Specificity

Distinguishes apoptotic cells from non-apoptotic cells in immunohistochemistry. Detects the p17 subunit of Caspase-3, but does not detect the precursor form in Western blot. We recommend R&D Systems Affinity Purified Anti-Caspase-3 (Catalog # AF-605-NA) or Monoclonal Anti-Cleaved-Caspase-3 (Catalog # MAB835) for Western blot applications.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Human/Mouse Active Caspase-3 Antibody

Caspase-3 antibody in Jurkat Human Cell Line by Immunocytochemistry (ICC).

Caspase-3 in Jurkat Human Cell Line.

Caspase-3 was detected in immersion fixed anti-FAS treated Jurkat human acute T cell leukemia cell line using 0.3 µg/mL Human/Mouse Active Caspase-3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF835) for 3 hours at room temperature. Cells were stained (red) and counterstained (green). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Caspase-3 antibody in Jurkat Human Cell Line by Immunocytochemistry (ICC).

Caspase‑3 in Jurkat Human Cell Line.

Caspase-3 was detected in immersion fixed Jurkat human acute T cell leukemia cell line stimulated with staurosporin using Human/Mouse Active Caspase-3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF835) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (yellow; Catalog # NL004) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Caspase-3 antibody in Human Colon Cancer Tissue by Immunohistochemistry (IHC-P).

Caspase‑3 in Human Colon Cancer Tissue.

Caspase-3 was detected in immersion fixed paraffin-embedded sections of human colon cancer tissue using Rabbit Anti-Human/Mouse Active Caspase-3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF835) at 1 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Applications for Human/Mouse Active Caspase-3 Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed anti-FAS treated or staurosporin-stimulated Jurkat human acute T cell leukemia cell line

Immunohistochemistry

1-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human colon cancer tissue

Reviewed Applications

Read 12 reviews rated 4.8 using AF835 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Reconstitution Buffer Available:
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Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Caspase-3

Caspase-3 (Cysteine-aspartic acid protease 3/Casp3; also Yama, apopain and CPP32) is a 29 kDa heterodimer that belongs to the peptidase C14A family of enzymes. It is widely expressed, and considered to be the major executioner caspase in the apoptotic cascade. Human procaspase-3 is a 32 kDa, 277 amino acid (aa) protein and is normally an inactive homodimer. Following cell stress/activation, procaspase-3 undergoes proteolysis to generate an N-terminal 148 aa p17/17 kDa subunit (aa 29-175), plus a 102 aa C-terminal p12/12 kDa subunit. These subunits noncovalently heterodimerize, and associate with another p17/p12 heterodimer to form an active enzyme. There is one potential variant that shows an alternative start site nine aa upstream of the standard start site coupled with a 21 aa substitution for aa 162-277. Over aa 29-175, human and mouse caspase-3 share 87% aa identity.

Alternate Names

Apopain, CASP3, Caspase3, CPP32, LICE-1, YAMA

Entrez Gene IDs

836 (Human); 12367 (Mouse)

Gene Symbol

CASP3

UniProt

Additional Caspase-3 Products

Product Documents for Human/Mouse Active Caspase-3 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human/Mouse Active Caspase-3 Antibody

For research use only

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