Anti-Caspase-3 p12 antibody [EPR16888]

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Caspase-3 p12 antibody [EPR16888] (AB179517)

Intracellular 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.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Caspase-3 p12 antibody [EPR16888] (AB179517)

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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-3 p12 antibody [EPR16888] (AB179517)

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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-3 p12 antibody [EPR16888] (AB179517)

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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-3 p12 antibody [EPR16888] (AB179517)

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.

• WB

Lab

Western blot - Anti-Caspase-3 p12 antibody [EPR16888] (AB179517)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. ab181602 was used as loading control. Bands between 27-32kDa represent cleavage of the procaspase at D9 and D28, respectively (PMID : 14567691)

All lanes:

Western blot - Anti-Caspase-3 p12 antibody [EPR16888] (ab179517) at 1/1000 dilution

Lane 1:

Mouse Alzheimer's disease brain tissue lysate at 20 µg

Lane 2:

Mouse brain cancer tissue lysate at 20 µg

Lane 3:

Mouse hippocampus tissue lysate at 20 µg

Lane 4:

Mouse spinal cord tissue lysate at 20 µg

Lane 5:

Mouse cerebellum tissue lysate at 20 µg

Lane 6:

Mouse cerebral cortex tissue lysate at 20 µg

Lane 7:

Mouse hypothalamus tissue lysate at 20 µg

Lane 8:

Mouse heart tissue lysate at 20 µg

Lane 9:

Mouse liver tissue lysate at 20 µg

Lane 10:

Human brain tissue lysate at 20 µg

Lane 11:

Human liver tissue lysate at 20 µg

Lane 12:

Human hypothalamus tissue lysate at 20 µg

Lane 13:

Human heart tissue lysate at 20 µg

Lane 14:

Human cerebellum tissue at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 32 kDa

false

Exposure time: 7s

• WB

Supplier Data

Western blot - Anti-Caspase-3 p12 antibody [EPR16888] (AB179517)

Blocking/Dilution buffer : 5% NFDM/TBST.

32 and 29 kDa bands represent procaspase-3; 12-kDa band is Caspase-3 subunit p12.

All lanes:

Western blot - 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 at 10 µg

Lane 2:

Untreated NIH/3T3 whole cell lysates at 10 µg

Secondary

All lanes:

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

Predicted band size: 32 kDa

Observed band size: 12 kDa,29 kDa,32 kDa

false

• WB

Supplier Data

Western blot - Anti-Caspase-3 p12 antibody [EPR16888] (AB179517)

Blocking/Dilution buffer : 5% NFDM/TBST.

32 and 29 kDa bands represent procaspase-3; 12-kDa band is Caspase-3 subunit p12.

All lanes:

Western blot - 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 at 10 µg

Lane 2:

HeLa untreated whole cell lysates at 10 µg

Secondary

All lanes:

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

Predicted band size: 32 kDa

Observed band size: 12 kDa,29 kDa,32 kDa

false

• WB

Supplier Data

Western blot - Anti-Caspase-3 p12 antibody [EPR16888] (AB179517)

Blocking/Dilution buffer : 5% NFDM/TBST.

32 and 29 kDa bands represent procaspase-3.

All lanes:

Western blot - Anti-Caspase-3 p12 antibody [EPR16888] (ab179517) at 1/5000 dilution

All lanes:

Mouse spleen lysates at 10 µg

Secondary

All lanes:

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

Predicted band size: 32 kDa

Observed band size: 29 kDa,32 kDa

false

• WB

Lab

Western blot - Anti-Caspase-3 p12 antibody [EPR16888] (AB179517)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. ab181602 was used as loading control. Bands between 27-32kDa represent cleavage of the procaspase at D9 and D28, respectively (PMID : 14567691)

All lanes:

Western blot - Anti-Caspase-3 p12 antibody [EPR16888] (ab179517) at 1/1000 dilution

Lane 1:

Rat brain tissue lysate at 20 µg

Lane 2:

Rat hippocampus tissue lysate at 20 µg

Lane 3:

Rat spinal cord tissue lysate at 20 µg

Lane 4:

Rat cerebellum tissue lysate at 20 µg

Lane 5:

Rat cerebral cortex tissue at 20 µg

Lane 6:

Rat hypothalamus tissue at 20 µg

Lane 7:

Rat heart tissue lysate at 20 µg

Lane 8:

Rat liver tissue lysate at 20 µg

Lane 9:

PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 32 kDa

false

Exposure time: 5s

• WB

Supplier Data

Western blot - Anti-Caspase-3 p12 antibody [EPR16888] (AB179517)

Blocking/Dilution buffer : 5% NFDM/TBST.

32 and 29 kDa bands represent procaspase-3.

All lanes:

Western blot - Anti-Caspase-3 p12 antibody [EPR16888] (ab179517) at 1/1000 dilution

Lane 1:

Mouse brain lysates at 10 µg

Lane 2:

Mouse heart lysates at 10 µg

Lane 3:

Rat brain lysates at 10 µg

Lane 4:

Rat heart lysates at 10 µg

Secondary

All lanes:

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

Predicted band size: 32 kDa

Observed band size: 29 kDa,32 kDa

false