Antibodies, Proteins, Kits and Reagents for Life Science

348 441 ₸ Product size

100 µl 348 441 ₸

Order now and get it on Tuesday March 09, 2021

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

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR18107] to Caspase-9
Suitable for: IHC-P, WB, ICC/IF, IP
Knockout validated
Reacts with: Mouse, Human

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 & Species

Application	Species
ICC/IF	Human
IHC-P	Human
IP	Human
WB	Mouse Human

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

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

pH: 7.2
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

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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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.
Anti-Caspase-9 antibody [EPR18107] (ab202068)

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