Caspase 3 p17 (Cleaved-Asp175) antibody

Catalog No. ABIN7251013

The Rabbit Polyclonal anti- antibody has been validated for WB, ELISA, IF and IHC (p) and has been independently validated. It is suitable to detect in samples from Human, Mouse and Rat.

Quick Overview for Caspase 3 p17 (Cleaved-Asp175) antibody (ABIN7251013)

Target: Caspase 3 p17

Reactivity: Human, Mouse, Rat

Host: Rabbit

Clonality: Polyclonal

Conjugate: Un-conjugated

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

Product Details

Binding Specificity: Cleaved-Asp175

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