Name: Anti-Caspase-9 antibody [EPR18107]
Brand: RabMAb
SKU: ab202068
Rating: 5 (2 reviews)

Product name

Anti-Caspase-9 antibody [EPR18107]
See all Caspase-9 primary antibodies
Description

Rabbit monoclonal [EPR18107] to Caspase-9
Host species

Rabbit
Tested applications

Suitable for: IHC-P, WB, ICC/IF, IPmore details
Species reactivity

Reacts with: Mouse, Human
Immunogen

Recombinant fragment within Human Caspase-9 aa 100-300. The exact sequence is proprietary.
Database link: P55211

Run BLAST with Run BLAST with
Positive control
- WB: HeLa and C2C12 whole cell lysates; Human fetal brain, fetal heart, fetal kidney and fetal liver lysates. IHC-P: Human cervix carcinoma tissue. ICC/IF: HeLa cells. IP: HeLa treated with staurosporine 1uM for 4 hours whole cell lysate.
General notes

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

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR18107
Isotype

IgG
Research areas

Alternative Versions
- Anti-Caspase-9 antibody [EPR18107] - BSA and Azide free (ab222231)
Assay kits
- Caspase 9 Assay Kit (Colorimetric) (ab65608)
- Caspase 9 (active) FITC Staining Kit (ab65615)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant human Caspase-9 protein (Tagged) (ab198061)

Our Abpromise guarantee covers the use of ab202068 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-P		1/300. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB		1/2000. Detects a band of approximately 46, 35, 37 kDa (predicted molecular weight: 46 kDa).
ICC/IF		1/500.
IP		1/80.

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.
Target information above from: UniProt accession P55211 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 842 Human
- Entrez Gene: 12371 Mouse
- Omim: 602234 Human
- SwissProt: P55211 Human
- SwissProt: Q8C3Q9 Mouse
- Unigene: 329502 Human
- Unigene: 88829 Mouse
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

Western blot - Anti-Caspase-9 antibody [EPR18107] (ab202068)

Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Caspase-9 knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: Jurkat cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab202068 observed at 46 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab202068 was shown to recognize Caspase-9 when Caspase-9 knockout samples were used, along with additional cross-reactive bands. Wild-type and Caspase-9 knockout samples were subjected to SDS-PAGE. ab202068 and ab8245 (loading control to GAPDH) were diluted at 1/2000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed withGoat anti-Rabbit IgG H&L (IRDy^e® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 h at room temperature before imaging.
Immunocytochemistry/ Immunofluorescence - Anti-Caspase-9 antibody [EPR18107] (ab202068)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Caspase-9 with ab202068 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).
Confocal image showing cytoplasmic and nuclear staining on HeLa cell line. The expression increased after treatment with staurosporine (1uM) for 4 hours.
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: ab202067 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-9 antibody [EPR18107] (ab202068)

Immunohistochemical analysis of paraffin-embedded Human cervix carcinoma tissue labeling Caspase-9 with ab202068 at 1/300 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.
Cytoplasmic and nuclear staining on Human cervix carcinoma tissue 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.
Western blot - Anti-Caspase-9 antibody [EPR18107] (ab202068)

Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Caspase-9 knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: Jurkat cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green).

Green - target observed at 46 kDa. Red - loading control, ab8245, observed at 37 kDa.

This western blot image is a comparison between ab202068 and a competitor's top cited rabbit polyclonal antibody.
Western blot - Anti-Caspase-9 antibody [EPR18107] (ab202068)

All lanes : Anti-Caspase-9 antibody [EPR18107] (ab202068) at 1/10000 dilution

Lane 1 : Untreated C2C12 (Mouse myoblast cell line) whole cell lysate
Lane 2 : C2C12 (Mouse myoblast cell line) treated with staurosporine 1uM for 4 hours whole cell lysate

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 46 kDa
Observed band size: 37,39,46 kDa
why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-Caspase-9 antibody [EPR18107] (ab202068)

All lanes : Anti-Caspase-9 antibody [EPR18107] (ab202068) at 1/2000 dilution

Lane 1 : Human fetal brain lysate
Lane 2 : Human fetal heart lysate

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 46 kDa
Observed band size: 46 kDa

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-Caspase-9 antibody [EPR18107] (ab202068)

All lanes : Anti-Caspase-9 antibody [EPR18107] (ab202068) at 1/2000 dilution

Lane 1 : Human fetal kidney lysate
Lane 2 : Human fetal liver lysate

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 46 kDa
Observed band size: 46 kDa

Exposure time: 30 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunoprecipitation - Anti-Caspase-9 antibody [EPR18107] (ab202068)

Caspase-9 was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) treated with staurosporine 1uM for 4 hours whole cell lysate with ab202068 at 1/80 dilution.
Western blot was performed from the immunoprecipitate using ab202068 at 1/1000 dilution.
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG was used as secondary antibody at 1/1500 dilution.
Lane 1: HeLa treated with staurosporine 1uM for 4 hours whole cell lysate10 µg (Input).
Lane 2: ab202068 IP in HeLa treated with staurosporine 1uM for 4 hours whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab202068 in HeLa treated with staurosporine 1uM for 4 hours whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 seconds.
Western blot - Anti-Caspase-9 antibody [EPR18107] (ab202068)

All lanes : Anti-Caspase-9 antibody [EPR18107] (ab202068) at 1/50000 dilution

Lane 1 : Untreated HeLa (human epithelial cells from cervix adenocarcinoma) whole cell lysate at 10 µg
Lane 2 : HeLa (human epithelial cells from cervix adenocarcinoma) treated with staurosporine 1 µM for 4 hours whole cell lysate at 20 µg

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 46 kDa
Observed band size: 35,37,46 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

This product has been referenced in:

Gao L et al. Dual inhibition of mTORC1/2 by DCZ0358 induces cytotoxicity in multiple myeloma and overcomes the protective effect of the bone marrow microenvironment. Cancer Lett 421:135-144 (2018). WB . Read more (PubMed: 29428642) »
Ding N et al. miR-378a-3p exerts tumor suppressive function on the tumorigenesis of esophageal squamous cell carcinoma by targeting Rab10. Int J Mol Med 42:381-391 (2018). Read more (PubMed: 29693138) »

See all 13 Publications for this product

Publishing research using ab202068? Please let us know so that we can cite the reference in this datasheet.

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1-2 of 2 Abreviews

Application

Western blot

Sample

Mouse Cell lysate - whole cell (B16 cells)

Gel Running Conditions

Reduced Denaturing (4-20% Tris Glycin gel, semi dry transfer)

Loading amount

50 µg

Specification

B16 cells

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Jun 18 2018

Application

Western blot

Sample

Human Cell lysate - whole cell (OV90 cells)

Gel Running Conditions

Reduced Denaturing (10% Tris Glycin gel)

Loading amount

50000 cells

Specification

OV90 cells

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Aug 15 2016

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
For licensing inquiries, please contact partnerships@abcam.com

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Recombinant Anti-Caspase-9 antibody [EPR18107] (ab202068)

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