| Field | Specification |
|---|---|
| Mfr No | |
| Clonality | |
| Concentration | |
| Conjugate | |
| Host | |
| Immunogen | Synthesized peptide derived from the Internal region of human Caspase-3 p17 |
| Isotype | |
| Product Type | |
| Shipping | |
| Storage | |
| UniProt # |
Product Overview
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 '216-Asp-Gly-217' bond. Cleaves and activates sterol regulatory element binding proteins (SREBPs) between the basic helix-loop-helix leucine zipper domain and the membrane attachment domain. Cleaves and activates caspase-6, -7 and -9. Involved in the cleavage of huntingtin.
Validated Applications & Recommended Dilutions
| Application | Recommended Dilution |
|---|---|
| WB | 1:500-2000 |
| IHC | 1:50-300 |
| IF | 1:50-300 |
Antibody Specifications
| Target | Cleaved-Caspase-3 p17 (D175) |
|---|---|
| Host Species | Rabbit |
| Clonality | Polyclonal |
| Isotype | IgG |
| Conjugate | Unconjugated |
| Purification | Affinity Purification |
| Concentration | 1 mg/mL |
| Molecular Weight | Calculated 32kDa; Observed 20kDa |
Immunogen
Synthesized peptide derived from the Internal region of human Caspase-3 p17 UniProt: P42574.
Species Reactivity & Cross-Reactivity
Validated for: Human, Mouse, Rat. Verify suitability in your sample type prior to use. For species or applications not listed in the datasheet, consult our technical team before purchase.
Storage & Handling
Store at -20℃ Valid for 12 months. Avoid freeze / thaw cycles.
Storage Buffer: PBS with 0.02% sodium azide, 0.5% protective protein and 50% glycerol, pH7.4
Related Products
Browse additional antibodies targeting Cleaved-Caspase-3 p17 (D175) and complementary reagents — including secondary antibodies, blocking buffers, and detection kits — available through BioHippo.
The recommended starting dilution for Western Blot is WB 1:500-2000;IHC 1:50-300;IF 1:50-300. Optimal dilution should be determined empirically for each laboratory setup, sample type, and detection system. Use freshly prepared lysates with protease inhibitors; add phosphatase inhibitors if studying phosphorylation-dependent forms of Cleaved-Caspase-3 p17 (D175). Expected band size: Calculated 32kDa, Observed 20kDa. If background is high, increase blocking (5% non-fat milk or BSA in TBST) and/or increase wash stringency.
For IHC-P, heat-induced epitope retrieval (HIER) is typically required. Recommended starting conditions: citrate buffer pH 6.0 (microwave or pressure cooker, ~20 min) for rodent samples; EDTA buffer pH 9.0 for human clinical samples. After retrieval, allow sections to cool gradually before proceeding to blocking. Include a Cleaved-Caspase-3 p17 (D175)-positive tissue control (confirmed by literature or database expression data) and a negative isotype control in every run. Recommended dilution: WB 1:500-2000;IHC 1:50-300;IF 1:50-300. Refer to the product datasheet for any target-specific retrieval guidance.
For intracellular targets including most nuclear and cytoplasmic proteins, fix cells with 4% paraformaldehyde (15 min, room temperature) followed by permeabilization with 0.1–0.2% Triton X-100 or saponin (5–10 min). For surface membrane targets, fixation without permeabilization may be sufficient. Block with 5–10% normal serum (matching the secondary antibody host species) for 30–60 min. Incubate primary antibody at the recommended dilution (WB 1:500-2000;IHC 1:50-300;IF 1:50-300) at 4°C overnight or 1–2 h at room temperature. Confirm nuclear/cytoplasmic localization of Cleaved-Caspase-3 p17 (D175) against published protein databases before designing your protocol.
This antibody has been experimentally validated in: Human, Mouse, Rat. Cross-reactivity with other species has not been systematically evaluated. If your sample species is not listed, assess immunogen sequence homology to your target species ortholog. The immunogen used is: "Synthesized peptide derived from the Internal region of human Caspase-3 p17" — higher sequence identity between species increases the probability of cross-reactivity, but experimental validation with appropriate controls is necessary before drawing scientific conclusions.
Store at -20℃ Valid for 12 months. Avoid freeze / thaw cycles. Storage Buffer: PBS with 0.02% sodium azide, 0.5% protective protein and 50% glycerol, pH7.4. To maintain antibody activity, avoid repeated freeze-thaw cycles. Aliquot into single-use volumes before first freeze. If working with the antibody frequently, keep a working stock at 4°C for up to 4 weeks — add 0.02% sodium azide to the working stock as a bacteriostatic agent if azide is not already present in the buffer. Monitor performance against a validated positive control when transitioning between storage aliquots or lots.
Polyclonal antibodies are produced from immunized animals and may exhibit some degree of lot-to-lot variation in titer, background, and sensitivity. Each production lot is tested for performance against established acceptance criteria. When transitioning to a new lot, run a side-by-side pilot comparison against your previous lot using a validated positive control sample before updating your standard protocol. If significant performance differences are observed, contact our technical support team — titer adjustments may resolve the issue in most cases.
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