Anti-Caspase-3 p12 antibody [EPR16888] (ab179517)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR16888] to Caspase-3 p12
- Suitable for: WB, IHC-P, ICC/IF, Flow Cyt
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-Caspase-3 p12 antibody [EPR16888]
See all Caspase-3 p12 primary antibodies -
Description
Rabbit monoclonal [EPR16888] to Caspase-3 p12 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt HumanICC/IF HumanIHC-P MouseRatHumanWB MouseHuman -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa and NIH/3T3 whole cell lysates. Mouse brain, heart and spleen; and Rat brain and heart lysates. IHC-P: Human tonsil, mouse testis and rat kidney tissues. ICC/IF: K562 cells.
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Properties
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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 (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR16888 -
Isotype
IgG
Images
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All lanes : Anti-Caspase-3 p12 antibody [EPR16888] (ab179517) at 1/1000 dilution
Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates treated with staurosporine 1uM for 4hr
Lane 2 : HeLa untreated whole cell lysates
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: 32, 29, 12 kDa
Observed band size: 12,29,32 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
32 and 29 kDa bands represent procaspase-3; 12-kDa band is Caspase-3 subunit p12.
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Flow Cytometry analysis of K562 (human chronic myelogenous leukemia) with purified ab179517 at 1/240 dilution (Red). The secondary antibody was Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution. A Rabbit monoclonal IgG (Black) was used as the isotype control and cells without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling Caspase-3 p12 with ab179517 at 1/150 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Confocal image showing cytoplasmic staining on K562 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows;
-ve control 1: ab179517 at 1/150 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution. -
All lanes : Anti-Caspase-3 p12 antibody [EPR16888] (ab179517) at 1/1000 dilution
Lane 1 : NIH/3T3 (Mouse embyro fibroblast cells) treated with staurosporine 1uM for 4h
Lane 2 : Untreated NIH/3T3 whole cell lysates
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: 32, 29, 12 kDa
Observed band size: 12,29,32 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
32 and 29 kDa bands represent procaspase-3; 12-kDa band is Caspase-3 subunit p12.
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Anti-Caspase-3 p12 antibody [EPR16888] (ab179517) at 1/5000 dilution + Mouse spleen lysates at 10 µg
Secondary
Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 32, 29, 12 kDa
Observed band size: 29,32 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
32 and 29 kDa bands represent procaspase-3.
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All lanes : Anti-Caspase-3 p12 antibody [EPR16888] (ab179517) at 1/1000 dilution
Lane 1 : Mouse brain lysates
Lane 2 : Mouse heart lysates
Lane 3 : Rat brain lysates
Lane 4 : Rat heart lysates
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: 32, 29, 12 kDa
Observed band size: 29,32 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
32 and 29 kDa bands represent procaspase-3.
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Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Caspase-3 p12 with ab179517 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) secondary antibody (ab97051) at 1/500 dilution. Cytoplasmic and nuclear staining in germinal center, weaker staining in the mantle zone (MZ) lymphocytes is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) secondary antibody (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling Caspase-3 p12 with ab179517 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) secondary antibody (ab97051) at 1/500 dilution. Nuclear and cytoplasmic staining on leydig cells and spermatogonial cells is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) secondary antibody (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling Caspase-3 p12 with ab179517 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) secondary antibody (ab97051) at 1/500 dilution. Granular staining on epithelium from kidney is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) secondary antibody (ab97051) at 1/500 dilution.
[1] Afshin Samali, Boris Zhivotovsky, Dean P. Jonesa, Sten Orreniusa. Detection of pro-caspase-3 in cytosol and mitochondria of various tissues (rat). FEBS Letters 431 (1998) 167-169.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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