Overview

  • Product name
    Anti-active Caspase 9 antibody
  • Description
    Rabbit polyclonal to active Caspase 9
  • Host species
    Rabbit
  • Specificity
    This antibody recognizes only the cleaved caspase 9 (37 kDa). It does not recognize full-length caspase 9 or any other caspase.
  • Tested applications
    Suitable for: IP, WBmore details
  • Species reactivity
    Reacts with: Human
  • Positive control
    • WB: Etoposide treated Jurkat whole cell lysate.
  • General notes


    Caspases are synthesized as inactive pro-enzymes that are processed to active form in cells undergoing apoptosis. Caspase 9 is an important member of the caspase family. Upon induction of apoptosis, Cytochrome c released from mitochondria associates with pro-caspase 9 (47 kDa) and Apaf-1. The complex processes pro-caspase 9 into a large subunit (37 kDa/17 kDa) and a small subunit (10 kDa). Cleaved caspase 9 further processes other caspases including caspase 3 and caspase 6, to initiate a caspase cascade leading to apoptosis. This affinity purified antibody recognizing the active forms of caspase 9 provides a new tool for identifying apoptotic cell populations in both tissue sections and cultured cells.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer
    Preservative: 0.02% Thimerosal (merthiolate)
    Constituents: PBS, 50% Glycerol, 0.5% BSA
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Primary antibody notes
    Caspases are synthesized as inactive pro-enzymes that are processed to active form in cells undergoing apoptosis. Caspase 9 is an important member of the caspase family. Upon induction of apoptosis, Cytochrome c released from mitochondria associates with pro-caspase 9 (47 kDa) and Apaf-1. The complex processes pro-caspase 9 into a large subunit (37 kDa/17 kDa) and a small subunit (10 kDa). Cleaved caspase 9 further processes other caspases including caspase 3 and caspase 6, to initiate a caspase cascade leading to apoptosis. This affinity purified antibody recognizing the active forms of caspase 9 provides a new tool for identifying apoptotic cell populations in both tissue sections and cultured cells.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab2324 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
IP Use a concentration of 10 - 20 µg/ml.
WB Use a concentration of 0.5 - 1 µg/ml.

Target

  • Database links
  • Alternative names
    • APAF 3 antibody
    • APAF3 antibody
    • Apoptotic protease activating factor 3 antibody
    • Apoptotic protease MCH 6 antibody
    • CASP9 antibody
    • Caspase 9 antibody
    • Caspase 9 apoptosis related cysteine peptidase antibody
    • Cleaved caspase 9 antibody
    • ICE LAP6 antibody
    • ICE like apoptotic protease 6 antibody
    • MCH6 antibody
    • PPP1R56 antibody
    • Protein phosphatase 1 regulatory subunit 56 antibody
    see all

Images

  • All lanes : Anti-active Caspase 9 antibody (ab2324)

    Lane 1 : Jurkat cell lysate, untreated
    Lane 2 : Jurkat cell lysate, treated with etoposide for 6 hours

    Observed band size: 37 kDa
    why is the actual band size different from the predicted?

References

This product has been referenced in:
  • Tang Q  et al. miR-223/Hsp70/JNK/JUN/miR-223 feedback loop modulates the chemoresistance of osteosarcoma to cisplatin. Biochem Biophys Res Commun 497:827-834 (2018). WB . Read more (PubMed: 29432736) »
  • Wang Z  et al. Long noncoding RNA maternally expressed gene 3 knockdown alleviates lipopolysaccharide-induced inflammatory injury by up-regulation of miR-203 in ATDC5 cells. Biomed Pharmacother 100:240-249 (2018). WB . Read more (PubMed: 29432995) »
See all 25 Publications for this product

Customer reviews and Q&As

1-10 of 12 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Methanol
Permeabilization
No

Dr. Kirk Mcmanus

Verified customer

Submitted Oct 14 2008

Abcam has not validated the combination of species/application used in this Abreview.
Application
Flow Cytometry
Sample
Human Cell (Peripheral blood mononuclear cell)
Permeabilization
Yes - BD Perm/Wash buffer
Gating Strategy
lymphocytes
Specification
Peripheral blood mononuclear cell
Preparation
Cell harvesting/tissue preparation method: Spin down, wash in BD Perm/Wash buffer 1x, fix, wash 2x, and stain with primary antibody 30 minutes.
Sample buffer: Buffer composition at analysis was < 0.09% sodium azide with FBS in phosphate buffered solution.
Fixation
Formaldehyde

Ms. Janine Sabino

Verified customer

Submitted Aug 20 2018

Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing
Sample
Mouse Tissue lysate - whole (liver)
Specification
liver
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 Dec 26 2014

Answer

Thank you for contacting us.

I have had a look at our catalogue and the naming of our antibodies is indeed confusing. We work with different collaborators and they have different ways of naming the antibodies. For example, some of them will name the antibody anti-active caspase 9 antibody if it has been proved to recognise the cleaved caspase 9. Others, will simply call the antibody anti-caspase 9 even if it has been shown to recognise the cleaved procaspase 9.

To help you choose a relevant antibody, I have gone through our catalogue and picked out the antibodies which should recognise the cleaved form of caspase 9:

ab133520 (https://www.abcam.com/ab133520)
ab63342 (https://www.abcam.com/ab63342)
ab61129 (https://www.abcam.com/ab61129)

I have also found some antibodies which will recognise the procaspase (46 kDa) or fully cleaved caspase (25 kDa) as well as the cleaved caspase protein (35 kDa):

ab25758 (https://www.abcam.com/ab25758)
ab258131 (https://www.abcam.com/ab258131)
ab115161 (https://www.abcam.com/ab115161)

Alternatively, I have found a lot in Japan of ab2324 different from the one you received. If you would like ab2324 as a replacement, please note that this may take some time to arrive.

I hope this information is helpful to you. Please let me know which resolution you would prefer.

Read More

Answer

Unfortunately, this product is not covered by our guarantee as it was bought over 6 months ago.

Given that I believe the protocol used should have given satisfactory results and that your customer has spent much time in the lab trying to resolve this problem using the Dotblot test, I will look into making an exception this time and extending our guarantee.

We no longer have this antibody ab2324 in stock so I am unable to offer you a free of charge replacement with another lot. I can however suggest ab77814 (https://www.abcam.com/ab77814) or ab69541 (https://www.abcam.com/ab69541) as replacements. Alternatively, I could issue your customer with a credit note.

Please let me know which outcome your customer would prefer and inform them that this is an exception that I am making and that for the purchase of future products, the guarantee will only apply to products bought within the 6 month period.

Read More

Question
Answer

Thank you for taking time to complete our questionnaire and for contacting us. I am sorry to hear this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.

Having reviewed the protocol details, I believe this product should have given satisfactory results.

I apologise for the inconvenience. According to our records, this product was ordered on 30/06/11 and you received lot GR21477-3. Could you confirm whether this information is correct?

In addition, can I ask how you stored this antibody?

If the product was purchased within the last 6 months, I would be pleased to offer you a free of charge replacement or credit note in compensation.

Thank you for your cooperation. I look forward to hearing from you.

Read More

Answer

Thank you for contacting us.

I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

I am attaching our questionnaire so that we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. Could you also resend the results in a jpeg format as our system does not allow us to read rar files?

This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.


I look forward to receiving your reply.

Read More
Application
Flow Cytometry
Sample
Human Cell (Jurkat)
Specification
Jurkat
Preparation
Cell harvesting/tissue preparation method: Spin down, wash in FACS buffer 1x, fix, wash 2x, and stain with primary antibody overnight.
Sample buffer: Buffer composition at analysis was 0.1% sodium azide with FBS in phosphate buffered solution.
Fixation
Paraformaldehyde
Permeabilization
Yes - Triton X-100 and NP40
Gating Strategy
Isolate cell population from plot of SSC-A / FSA-A

Abcam user community

Verified customer

Submitted Sep 15 2011

Answer

Thank you for your enquiry. Pro-caspase-9 is an approximately 47 kDa protein that is cleaved at an autoactivation site 315/316 resulting in a 35 kDa fragment and a 12 kDa fragment. ab2324 will recognize the cleaved caspase-9 N-terminal fragment (p35)only. ab4824 will recognize cleavage site 315/316 and the C-terminal fragment (p12) only, and there is a peptide that you can buy for blocking (ab5251) to use as a positive/negative control for this antibody. ab19425 will recognize Pro-Caspase-9 (p47) and may or may not recognize p35. I hope this information helps, please do not hesitate to contact us if you need any more advice or information,

Read More

Answer

The immunogens are synthetic peptides mapping to the N-terminal of the cleavage site: Asp330 (human) and Asp353 (rat). Unfortunately, the exact sequence of the immunogen remains proprietary.

Read More

1-10 of 12 Abreviews or Q&A

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

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