Overview

  • Product name
    Anti-Cleaved Caspase-3 antibody
    See all Cleaved Caspase-3 primary antibodies
  • Description
    Rabbit polyclonal to Cleaved Caspase-3
  • Host species
    Rabbit
  • Specificity
    The antibody preferentially recognizes the p17 fragment of the active Caspase-3. Under the conditions we tested, the antibody did not detect the precursor form. Under other conditions (such as those used in PMC2206181), this antibody has been found to detect both the pro and active forms of Caspase-3. Although some customers have used this antibody successfully in mouse and rat, we do not batch test in these species and so cannot guarantee that it will work in them.
  • Tested applications
    Suitable for: IHC-FoFr, IHC-P, IHC-Fr, WBmore details
  • Species reactivity
    Reacts with: Rabbit, Human, Quail
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide corresponding to Human Cleaved Caspase-3. Synthetic peptide mapping to the N-terminus adjacent to the cleavage site of human caspase-3
    Database link: P42574
    (Peptide available as ab38283)

  • Positive control
    • Camptothecin (2 µM) treated Jurkat cells. If no signal is observed a time course may be required to identify maximal caspase activity.
  • General notes

    Caspases are synthesized as inactive pro-enzymes that are processed to active form in cells undergoing apoptosis. Caspase 3 has been extensively studied and implicated to play an important role in apoptosis. Active caspase 3 proteolytically cleaves and activates other caspases, as well as relevant targets in the cells (e.g., PARP). This affinity purified antibody recognizing the active forms of caspase-3 provides a new tool for identifying apoptotic cell populations in both tissue sections and cultured cells.

    Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).

    See other anti-rabbit secondary antibodies that can be used with this antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab2302 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-FoFr Use at an assay dependent concentration. PubMed: 18797916
IHC-P Use a concentration of 5 - 20 µg/ml. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.
IHC-Fr Use a concentration of 10 - 20 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 17 kDa.Can be blocked with Human Caspase-3 peptide (ab38283).

Target

  • Function
    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. Triggers cell adhesion in sympathetic neurons through RET cleavage.
  • Tissue specificity
    Highly expressed in lung, spleen, heart, liver and kidney. Moderate levels in brain and skeletal muscle, and low in testis. Also found in many cell lines, highest expression in cells of the immune system.
  • Sequence similarities
    Belongs to the peptidase C14A family.
  • Post-translational
    modifications
    Cleavage by granzyme B, caspase-6, caspase-8 and caspase-10 generates the two active subunits. Additional processing of the propeptides is likely due to the autocatalytic activity of the activated protease. Active heterodimers between the small subunit of caspase-7 protease and the large subunit of caspase-3 also occur and vice versa.
    S-nitrosylated on its catalytic site cysteine in unstimulated human cell lines and denitrosylated upon activation of the Fas apoptotic pathway, associated with an increase in intracellular caspase activity. Fas therefore activates caspase-3 not only by inducing the cleavage of the caspase zymogen to its active subunits, but also by stimulating the denitrosylation of its active site thiol.
  • Cellular localization
    Cytoplasm.
  • Information by UniProt
  • Database links
  • Alternative names
    • active caspase 3 antibody
    • Apopain antibody
    • CASP 3 antibody
    • CASP-3 antibody
    • CASP3 antibody
    • CASP3_HUMAN antibody
    • Caspase 3 antibody
    • Caspase-3 subunit p12 antibody
    • CPP 32 antibody
    • CPP-32 antibody
    • CPP32B antibody
    • Cysteine protease CPP32 antibody
    • PARP cleavage protease antibody
    • Protein Yama antibody
    • SCA-1 antibody
    • SCA1 antibody
    • SREBP cleavage activity 1 antibody
    • Yama antibody
    see all

Images

  • IHC image of cleaved caspase-3 staining in human tonsil formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab2302, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

     

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • All lanes : Anti-Cleaved Caspase-3 antibody (ab2302) at 1 µg/ml

    Lane 1 : HeLa Whole Cell Lysate (2 uM Staurosporine, 4Hr) at 20 µg
    Lane 2 : HeLa Whole Cell Lysate (untreated) at 20 µg
    Lane 3 : Cleaved Caspase 3 (recombinant protein) at 0.1 µg

    Secondary
    All lanes : 800CW Goat Anti-Rabbit IgG at 1/10000 dilution

    Performed under reducing conditions.

    Additional bands at: 17 kDa (possible mature (processed) protein), 24 kDa, 35 kDa. We are unsure as to the identity of these extra bands.

  • Immunohistochemistical detection (on formaldehyde/PFA-fixed paraffin-embedded sections) of cleaved Caspase-3 antibody (ab2302) on Quail Tissue sections (Quail E6/7 developing DRGs Saggital section). Antigen retrieval step: Heat mediated. Blocking step: 1% BSA for 10 mins RT. Primary Antibody ab2302 incubated at 1/50 for at RT. Secondary Antibody: Biotin conjugated goat anti rabbit IgG (1/300).

  • ab2302 staining cleaved Caspase-3 in human ulcerative colitis, colon tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed using the HOPE method and permeabilized. Samples were incubated with primary antibody (1/20) for 1 hour at 25°C. A Biotin-SP-conjugated donkey anti-rabbit IgG polyclonal (1/800) was used as the secondary antibody.

    See Abreview

  • Anti-Cleaved Caspase-3 antibody (ab2302) at 1 µg/ml + Camptothecin
    (2 µM) treated Jurkat cells.
  • Lung sections were stained with caspase antibody ab2302. Caspase-3 positive cells are shown in brown (DAB). This picture was kindly supplied as part of a customer review.

References

This product has been referenced in:
  • Chen Z  et al. Protective effects of lncRNA H19 silence against hypoxia-induced injury in PC-12 cells by regulating miR-28. Int J Biol Macromol 121:546-555 (2019). Read more (PubMed: 30312698) »
  • Mei F  et al. Magnesium isoglycyrrhizinate alleviates liver injury in obese rats with acute necrotizing pancreatitis. Pathol Res Pract 215:106-114 (2019). Read more (PubMed: 30396756) »
See all 306 Publications for this product

Customer reviews and Q&As

1-10 of 84 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (pancreas)
Antigen retrieval step
Heat mediated
Permeabilization
Yes - TBS, 0.4% TX100
Specification
pancreas
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jan 17 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (cartilage/bone)
Antigen retrieval step
Enzymatic - Buffer/Enzyme Used: 100ug/ml proteinase K
Permeabilization
No
Specification
cartilage/bone
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Jan 17 2019

Application
Western blot
Sample
Human Cell lysate - whole cell (neuroblastoma)
Gel Running Conditions
Reduced Denaturing (12)
Loading amount
40 µg
Specification
neuroblastoma
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Dec 21 2018

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Fibroblast from human tumor tissue)
Permeabilization
Yes - 0.1%Triton X100 in PBS
Specification
Fibroblast from human tumor tissue
Blocking step
DakoCytomation x0909 as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: RT°C
Fixative
Formaldehyde

Hongwei Shao

Verified customer

Submitted Dec 17 2018

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (mouse melanoma cells injected into mouse brain)
Antigen retrieval step
None
Permeabilization
Yes - Triton-X
Specification
mouse melanoma cells injected into mouse brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 24°C
Fixative
Paraformaldehyde

Dr. Yona Goldshmit

Verified customer

Submitted Dec 12 2018

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (neurons)
Permeabilization
Yes - triton X 100 1%
Specification
neurons
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Dec 11 2018

Application
Immunohistochemistry free floating
Sample
Rat Tissue sections (Brain)
Specification
Brain

Abcam user community

Verified customer

Submitted Oct 31 2018

Application
Immunohistochemistry free floating
Sample
Mouse Tissue sections (Brain)
Specification
Brain

Abcam user community

Verified customer

Submitted Oct 31 2018

Application
IHC - Wholemount
Sample
Rat Tissue (embryonic lung explant)
Specification
embryonic lung explant

Abcam user community

Verified customer

Submitted Jul 30 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (4T1 tumor)
Antigen retrieval step
Heat mediated
Permeabilization
No
Specification
4T1 tumor
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 25°C
Fixative
Formaldehyde

Dr. Galoz Kaneti

Verified customer

Submitted Jul 16 2018

1-10 of 84 Abreviews or Q&A

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