Anti-Caspase-3 p12 antibody [EPR16888] - BSA and Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Caspase-3 p12 antibody [EPR16888] - BSA and Azide free (AB238936)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179517).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Caspase-3 p12 antibody [EPR16888] - BSA and Azide free (AB238936)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179517).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-3 p12 antibody [EPR16888] - BSA and Azide free (AB238936)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179517).

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] - BSA and Azide free (AB238936)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179517).

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] - BSA and Azide free (AB238936)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179517).

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] - BSA and Azide free (AB238936)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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] (<a href='/en-us/products/primary-antibodies/caspase-3-p12-antibody-epr16888-ab179517'>ab179517</a>) 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

Lab

Western blot - Anti-Caspase-3 p12 antibody [EPR16888] - BSA and Azide free (AB238936)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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] (<a href='/en-us/products/primary-antibodies/caspase-3-p12-antibody-epr16888-ab179517'>ab179517</a>) 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