Overview

  • Product name

  • Description

    Rabbit polyclonal to Caspase-9
  • Host species

    Rabbit
  • Specificity

    Detects bands of ~46-50kDa (proform) and ~35/37kDa, ~18kDa, ~10kDa (processed) by Western blot.
  • Tested applications

    Suitable for: WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Sheep, Chicken, Guinea pig, Cow, Dog, Human, Monkey
  • Immunogen

    Recombinant fragment corresponding to Human Caspase-9 (N terminal). procaspase-9 region
    Database link: P55211

  • Positive control

    • HeLa Cell Lysate (Heat Shocked); Jurkat cell lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab69514 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 1/500. Detects a band of approximately 50 kDa (predicted molecular weight: 46 kDa).

Target

  • Function

    Involved in the activation cascade of caspases responsible for apoptosis execution. Binding of caspase-9 to Apaf-1 leads to activation of the protease which then cleaves and activates caspase-3. Proteolytically cleaves poly(ADP-ribose) polymerase (PARP).
    Isoform 2 lacks activity is an dominant-negative inhibitor of caspase-9.
  • Tissue specificity

    Ubiquitous, with highest expression in the heart, moderate expression in liver, skeletal muscle, and pancreas. Low levels in all other tissues. Within the heart, specifically expressed in myocytes.
  • Sequence similarities

    Belongs to the peptidase C14A family.
    Contains 1 CARD domain.
  • Developmental stage

    Expressed at low levels in fetal heart, at moderate levels in neonate heart, and at high levels in adult heart.
  • Post-translational
    modifications

    Cleavages at Asp-315 by granzyme B and at Asp-330 by caspase-3 generate the two active subunits. Caspase-8 and -10 can also be involved in these processing events.
  • Information by UniProt
  • Database links

  • Alternative names

    • APAF-3 antibody
    • APAF3 antibody
    • Apoptosis related cysteine peptidase antibody
    • Apoptotic protease Mch-6 antibody
    • Apoptotic protease-activating factor 3 antibody
    • CASP-9 antibody
    • CASP9 antibody
    • CASP9_HUMAN antibody
    • Caspase 9 apoptosis related cysteine peptidase antibody
    • Caspase 9 Dominant Negative antibody
    • Caspase 9c antibody
    • Caspase-9 antibody
    • Caspase-9 subunit p10 antibody
    • ICE LAP6 antibody
    • ICE like apoptotic protease 6 antibody
    • ICE-LAP6 antibody
    • ICE-like apoptotic protease 6 antibody
    • MCH6 antibody
    • PPP1R56 antibody
    • protein phosphatase 1, regulatory subunit 56 antibody
    • RNCASP9 antibody
    see all

Images

  • All lanes : Anti-Caspase-9 antibody (ab69514) at 1/500 dilution

    Lane 1 : Jurkat cell lysate
    Lane 2 : apoptosis-induced Jurkat cell lysate

    Predicted band size: 46 kDa
    Observed band size: 50 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 36 kDa, 40 kDa. We are unsure as to the identity of these extra bands.

References

This product has been referenced in:

  • Dai YJ  et al. Chronic inflammation is a key to inducing liver injury in blunt snout bream (Megalobrama amblycephala) fed with high-fat diet. Dev Comp Immunol 97:28-37 (2019). Read more (PubMed: 30910418) »
  • Yang Y  et al. Reduction in milk fat globule-EGF factor 8 inhibits triple-negative breast cancer cell viability and migration. Oncol Lett 17:3457-3465 (2019). Read more (PubMed: 30867784) »
See all 10 Publications for this product

Customer reviews and Q&As

Question

One of the customer bought ab44289/Mouse monoclonal [mAbcam 44289] to CNPase for IF assay, but there is no signal. Please help, thank you!  Code:ab69514, lot:445013, order reference No.872211 PO#110427-3 Customer:  The following are the details of the experiments.  1. Order details: Antibody storage conditions (temperature/reconstitution etc) Per 50μl PBS added 1μl antibody and then aliquoted and stored at -20°C 2. Please describe the problem (high background, no staining etc).           no signal 3. On what material are you testing the antibody in IHC *          Species: rat *          Cell Iine or tissue: frozen section of rat brain, each slice is 10μm   4. How did you fix the samples? Paraformaldehyde: 4% Fixation time: 30min Fixation temperature: RT Rupture membranes with 0.3%Triton-100 at RT for 30min   5.  For formalin-fixed paraffin embedded tissue: did you apply antigen retrieval step?             no 6.      Blocking steps:  Use 10% goat’s serum block the unspecific binding sites for 1.5h   7. Primary antibody Specification (in which species was it raised against): rat *          At what dilution(s) have you tested this antibody: 1:100   1:250    1:500   1:1000 *          What dilution buffer was used: 0.01M  PBS (contains 0.3%Triton-100) *          Incubation time: overnight *          Incubation temperature: 4℃ *          What washing steps were done: 0.01M  PBS wash, 3*3min 8. Secondary antibody Specification (in which species was it raised against): goat anti-mouse, IgM-TRITC,  No.1021-03 , from southern biotech At what dilution(s) have you tested this antibody: 1:250 *          What dilution buffer was used: 0.01M PBS *          Incubation time: 1.5h *          Incubation temperature: RT *          What washing steps were done: 0.01M  PBS wash, 3*3min *          Do you know whether the problems you are experiencing come from the secondary?  Yes, there is positive result, please see attachment(result) .The antibody( No. MAB1326)is from R&D company   9. What detection method are you using? IF   10. Background staining Picture *          Please provide an image of your staining     There is no signal in the result, so customer didn’t supply.   12. Did you apply positive and negative controls along with the samples? Please specify. no 13. Optimization attempts *          How many times have you tried the IHC?   Four times *          Do you obtain the same results every time?  Yes What steps have you altered? Omit  4%paraformaldehyde fixation; change the concentration of the antibody; extend the time of rupturing membranes    

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Answer

Thank you for providing the completed questionnaire form. I will be happy to help you resolve any issues you are having with ab44289. I just have a few comments: 1. It seems for storage you have diluted ab44289 50x in PBS before freezing? Is this correct? If so please note we don't recommend diluting the antibody for storage as antibodies become increasingly unstable at lower concentrations. 2. The secondary noted is goat anti-mouse, IgM-TRITC, No.1021-03 , from southern biotech. This secondary antibody is designed to detect mouse IgM but will not react with mouse IgG. ab44289 is an IgG molecule. I would recommend a secondary antibody such as Donkey polyclonal Secondary Antibody to Mouse IgG - H&L (TRITC ) (ab6817) which will react with mouse IgG. I hope this is helpful. Please contact me again if you have any further questions.

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