Overview

  • Product name
  • Description
    Rabbit polyclonal to Caspase-3
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Rabbit, Chicken, Guinea pig, Hamster, Cow, Dog, Human, Pig, Saccharomyces cerevisiae, Monkey
  • Immunogen

    Recombinant full length protein corresponding to Human Caspase-3 aa 1-277.
    Sequence:

    MENTENSVDSKSIKNLEPKIIHGSESMDSGISLDNSYKMDYPEMGLCIII NNKNFHKSTGMTSRSGTDVDAANLRETFRNLKYEVRNKNDLTREEIVELM RDVSKEDHSKRSSFVCVLLSHGEEGIIFGTNGPVDLKKITNFFRGDRCRS LTGKPKLFIIQACRGTELDCGIETDSGVDDDMACHKIPVEADFLYAYSTA PGYYSWRNSKDGSWFIQSLCAMLKQYADKLEFMHILTRVNRKVATEFESF SFDATFHAKKQIPCIVSMLTKELYFYH


    Database link: P42574

  • Positive control
    • Jurkat whole cell lysate (ab7899), Jurkat cells treated with staurosporine (lysate), HeLa whole cell lysate (ab150035)

Properties

Applications

Our Abpromise guarantee covers the use of ab90437 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
WB 1/1000. Predicted molecular weight: 32 kDa.

A lower molecular weight band refers to a proteolytic cleavage product of caspase-3.

IHC-P Use at an assay dependent concentration.

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.
  • 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
    • A830040C14Rik antibody
    • Apopain antibody
    • CASP-3 antibody
    • CASP3 antibody
    • CASP3_HUMAN antibody
    • Casp3a antibody
    • Caspase 3 antibody
    • Caspase 3, apoptosis-related cysteine peptidase antibody
    • Caspase 3, apoptosis-related cysteine protease antibody
    • Caspase 3, apoptosis-related cysteine protease a antibody
    • Caspase-3 subunit p12 antibody
    • CC3 antibody
    • CPP-32 antibody
    • CPP32 antibody
    • CPP32B antibody
    • Cysteine protease CPP32 antibody
    • EC 3.4.22.56 antibody
    • LICE antibody
    • mldy antibody
    • OTTHUMP00000165052 antibody
    • OTTHUMP00000165053 antibody
    • OTTHUMP00000165054 antibody
    • PARP cleavage protease antibody
    • Procaspase3 antibody
    • Protein Yama antibody
    • SCA 1 antibody
    • SCA-1 antibody
    • SREBP cleavage activity 1 antibody
    • Yama antibody
    see all

Images

  • All lanes : Anti-Caspase-3 antibody (ab90437) at 1/1000 dilution

    Lane 1 : Mouse hepatocytes whole cell lysate poly(I:C) treated - 0 hours
    Lane 2 : Mouse hepatocytes whole cell lysate poly(I:C) treated - 12 hours

    Developed using the ECL technique.

    Predicted band size: 32 kDa
    Observed band size: 32 kDa


    Exposure time: 20 seconds


    Blocking with 5% milk for 1 hour at 23°C.

    See Abreview

  • Immunohistochemistry analysis of human spleen tissue stained with Caspase-3, pAb at 10µg/ml.

  • All lanes : Anti-Caspase-3 antibody (ab90437) at 1/1000 dilution

    Lane 1 : Jurkat cell lysate
    Lane 2 : Jurkat cells treated with staurosporine
    Lane 3 : MCF-7 cell lysate (negative control)

    Developed using the ECL technique.

    Predicted band size: 32 kDa
    Additional bands at: ~18 kDa (possible cleavage fragment), ~20 kDa (possible cleavage fragment)



    Western blot analysis of Caspase-3 pAb.

References

This product has been referenced in:
  • Zhao S  et al. High-pressure carbon dioxide pneumoperitoneum induces oxidative stress and mitochondria-associated apoptotic pathway in rabbit kidneys with severe hydronephrosis. Int J Mol Med 43:305-315 (2019). Read more (PubMed: 30431064) »
  • Gong G  et al. Tanshinone IIA alleviates oxidative damage after spinal cord injury in vitro and in vivo through up-regulating miR-124. Life Sci 216:147-155 (2019). Read more (PubMed: 30468834) »
See all 41 Publications for this product

Customer reviews and Q&As

1-10 of 15 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (jurkat cell line)
Gel Running Conditions
Reduced Denaturing (4-12% bis-tris gel)
Loading amount
10 µg
Treatment
cytochrome c
Specification
jurkat cell line
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Mar 28 2019

Application
Western blot
Sample
Mouse Tissue lysate - whole (mouse RPE/choroid)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Treatment
cigarette smoking 6 mo
Specification
mouse RPE/choroid
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Mar 13 2019

Application
Immunohistochemistry (Frozen sections)
Sample
Chicken Tissue sections (Neural tube)
Permeabilization
No
Specification
Neural tube
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde

Ayelet Kohl

Verified customer

Submitted Sep 21 2016

Application
Western blot
Sample
Rat Tissue lysate - whole (Skeletal muscle)
Gel Running Conditions
Reduced Denaturing (15%)
Loading amount
20 µg
Specification
Skeletal muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Feb 29 2016

Application
Western blot
Sample
Mouse Tissue lysate - whole (Skeletal muscle)
Gel Running Conditions
Reduced Denaturing (15%)
Loading amount
20 µg
Specification
Skeletal muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Feb 29 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Chicken Cell (Embryo)
Permeabilization
Yes - 0.1%Tween
Specification
Embryo
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Formaldehyde

Maneeshi Prasad

Verified customer

Submitted Oct 08 2015

Application
Western blot
Sample
Mouse Cell lysate - whole cell (mouse hepatocytes)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Treatment
20 ug/m poly(I:C)
Specification
mouse hepatocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Sep 01 2015

Answer



Here are answers to your questions:

1. Since the immunogen is full length and the antibody is polyclonal this antibody will react with all forms of casepase 3.

2. The 2 bands with lower molecular weights below 22kDa are p17 and p12 which appear after processing of Caspase 3.

Read More
Application
Western blot
Sample
Mouse Tissue lysate - whole (Skeletal muscle (AT))
Loading amount
50 µg
Specification
Skeletal muscle (AT)
Gel Running Conditions
Reduced Denaturing (15% gel)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Dr. G.K Sakellariou

Verified customer

Submitted Sep 28 2012

Question
Answer

Thank you for your reply.

There are a few things I would suggest may help improve the results:

1. I would suggest loading a higher quantity onto your gel, a good starting point would be 20 ug/lane.

2. I would suggest not using different blocking agents in the blocking buffer and the antibody diluent. I.e. I would try 5% BSA TBST blocking with 5% BSA TBST diluent of the primary antibody and separately 3% milk blocking and milk in the antibody diluent. I realise that the Abreview that has been submitted with this antibody has used a mixture of the two but we would not typically recommend this as it can lead to a higher background.

In order to understand the problems you have been encountering, could you explain more fully what the problems have been? I.e. high background, no bands etc?

If possible, could you share an image of the results with me?

I look forward to receiving your reply.

Read More

1-10 of 15 Abreviews or Q&A

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