Caspase 3 p17 (Cleaved-Asp175) antibody

Catalog No. ABIN7251013

Product Details

Target: Caspase 3 p17

Binding Specificity: Cleaved-Asp175

Reactivity: Human, Mouse, Rat

Host: Rabbit

Clonality: Polyclonal

Conjugate: Un-conjugated

Application: Western Blotting (WB), ELISA, Immunofluorescence (IF), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))

Characteristics: Polyclonal Antibody

Purification: Affinity purification

Immunogen: Synthesized peptide derived from the Internal region of human Caspase-3 p17

Isotype: IgG

Application Details

Application Notes: WB 1:500-2000, IHC 1:50-300, IF 1:50-300

Restrictions: For Research Use only

Independent Validation (IHC)

Validation #104629 (Immunohistochemistry)

by: Prof. Merighi, Laboratory of Neurobiology, Department of Veterinary Sciences, University of Turin

No.: #104629

Date: 03/15/2025

Antigen: Caspase 3 p17

Lot Number: DX03VBDV9989

Method validated: Immunohistochemistry

Positive Control:

The expression of caspase 3 at P5 was already attested in our laboratory (Lossi et al., 2004).

Negative Control:

A control slice was processed for each experimental procedure, omitting the primary antibody.

Notes:

Reference: Lossi L., Tamagno I. and Merighi A. (2004) “Molecular morphology of neuronal apoptosis: analysis of Caspase 3 activation during postnatal development of mouse cerebellar cortex.” J Mol Histol, 35(6):621-9.

Validation Images

Expression of cleaved caspase 3 (cCASP3) in P6 mouse cerebellum. An immunoreactive Purkinje neuron (left) and two cCASP3-positive granule cells (right) after immunostaining with ABIN7251013 and nuclear counterstaining with 4′,6-Diamidine-2′-phenylindole dihydrochloride (DAPI). EGL = external granular layer of the cerebellar cortex; IGL = internal granular layer of the cerebellar cortex.

Full Methods

Primary Antibody: ABIN7251013

Secondary Antibody: anti-rabbit Alexa Fluor 488 (Invitrogen. Lot 2541675)

Full Protocol:

• Sample analyzed: mouse cerebellum at post-natal day (P) 5–6.
• Paraffin-embedded slices (7 µm thick) were deparaffinized and rehydrated through a graded series of alcohols and distilled water (dH₂O).
• Sections were blocked in ovalbumin 1% 1 hour at room temperature.
• Antigen retrieval was performed in all the sections by microwave treatment (1 minute at 750 W + 1 minute at 250 W in citrate buffer pH 6).
• 3×5 minutes washing in 0.01 M PBS.
• Sections were incubated with the primary antibody at the following dilutions of 1:50/1:100/1:200, overnight at room temperature (primary antibody was diluted in PLL/BSA/PBS diluent).
• 3×5 minutes washing in 0.01 M PBS.
• Incubation with the anti-rabbit secondary antibody 1:400 in PLL/BSA/PBS diluent, 1 hour at room temperature.
• 3×5 minutes washing in 0.01 M PBS.
• Nuclear counterstaining was performed with DAPI 1 µg/mL, 2 minutes.
• 2×5 minutes washing in 0.01 M PBS.
• Specimens were then mounted in Fluoroshield mounting medium (Sigma).

Experimental Notes:

Passed. The antibody works in IHC-P at concentrations from 1:50-1:200

Handling

Format: Liquid

Concentration: 1 mg/mL

Buffer: PBS with 0.02 % sodium azide, 0.5 % BSA and 50 % glycerol, pH 7.4

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: -20 °C

Storage Comment: Store at -20°C. Avoid freeze / thaw cycles.

Target Details

Target: Caspase 3 p17

Alternative Name: CASP3 p17

Background: 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.

Molecular Weight:

Observed_MW: 20 kDa

Calculated_MW: 32 kDa

UniProt: P42574