Recombinant
RabMAb

Anti-Caspase-6 antibody [EPR18043] (ab185645)

Overview

  • Product name
    Anti-Caspase-6 antibody [EPR18043]
    See all Caspase-6 primary antibodies
  • Description
    Rabbit monoclonal [EPR18043] to Caspase-6
  • Tested applications
    Suitable for: WB, IHC-P, IP, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human Caspase-6 aa 200 to the C-terminus. The exact sequence is proprietary.
    Database link: P55212

  • Positive control
    • WB: Jurkat, NIH/3T3, C6 and PC-12 whole cell lysates. IHC-P: Human colon, mouse brain and rat brain tissues. ICC/IF: Jurkat cells. IP: NIH/3T3 whole cell lysate treated with 1uM staurosporine for 4 hours.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

Properties

Applications

Our Abpromise guarantee covers the use of ab185645 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. Detects a band of approximately 33, 11 kDa (predicted molecular weight: 33 kDa).
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP 1/50.
ICC/IF 1/500.

Target

  • Function
    Involved in the activation cascade of caspases responsible for apoptosis execution. Cleaves poly(ADP-ribose) polymerase in vitro, as well as lamins. Overexpression promotes programmed cell death.
  • Sequence similarities
    Belongs to the peptidase C14A family.
  • Post-translational
    modifications
    Cleavages by caspase-3, caspase-8 or -10 generate the two active subunits.
  • Cellular localization
    Cytoplasm.
  • Information by UniProt
  • Database links
  • Alternative names
    • Apoptotic protease Mch-2 antibody
    • Apoptotic protease MCH2 antibody
    • CASP-6 antibody
    • CASP6 antibody
    • CASP6_HUMAN antibody
    • Caspase 6 antibody
    • Caspase 6 apoptosis related cysteine protease antibody
    • Caspase 6, apoptosis related cysteine peptidase antibody
    • Caspase-6 subunit p11 antibody
    • Mch2 antibody
    • OTTHUMP00000162742 antibody
    • OTTHUMP00000162743 antibody
    see all

Images

  • All lanes : Anti-Caspase-6 antibody [EPR18043] (ab185645) at 1/1000 dilution

    Lane 1 : Untreated Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate
    Lane 2 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate treated with 1uM staurosporine for 4 hours

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size : 33 kDa
    Observed band size : 11,33 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 minute

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-Caspase-6 antibody [EPR18043] (ab185645) at 1/1000 dilution

    Lane 1 : Untreated NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate
    Lane 2 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate treated with 1uM staurosporine for 4 hours

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size : 33 kDa
    Observed band size : 11,33 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 minute

    The predicted MW is 32kDa for mouse and rat full-length procaspase-6.

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-Caspase-6 antibody [EPR18043] (ab185645) at 1/1000 dilution

    Lane 1 : C6 (Rat glial tumor cells) whole cell lysate
    Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size : 33 kDa
    Observed band size : 33 kDa


    Exposure time : 1 minute

    The predicted MW is 32kDa for mouse and rat full-length procaspase-6.

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling Caspase-6 with ab185645 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).

    Confocal image showing both nuclear and cytoplasmic staining on Jurkat cells. The staining remained similar after treatment with staurosporine (1uM, 4 hours) as the antibody interacts with the subunit p11. 

    The nuclear counter stain is DAPI (blue).

    Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
    The negative controls are as follows:
    -ve control 1: ab185645 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Caspase-6 with ab185645 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

    Cytoplasmic and nuclear staining on epithelial cells of human colon is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded Mouse brain tissue labeling Caspase-6 with ab185645 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

    Cytoplasmic staining on neurons of mouse brain is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded Rat brain tissue labeling Caspase-6 with ab185645 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

    Cytoplasmic staining on neurons of rat brain is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Caspase-6 was immunoprecipitated from 1mg of NIH/3T3 (Mouse embyro fibroblast cells) treated with 1uM staurosporine for 4 hours whole cell lysate with ab185645 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab185645 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used as secondary antibody at 1/1500 dilution.

    Lane 1: NIH/3T3 treated with 1uM staurosporine for 4 hours whole cell lysate10 µg (Input). Lane 2: ab185645 IP in NIH/3T3 treated with 1uM staurosporine for 4 hours whole cell lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab185645 in NIH/3T3 treated with 1uM staurosporine for 4 hours whole cell lysate.

    The cleaved Caspase-6 appears slightly larger than 11kDa. This fragment contains subunit p11 plus the internal propeptide.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

References

ab185645 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Tonsil)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Tonsil
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

Submitted Feb 13 2017

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up