Anti-Protein CASP antibody [EPR18806]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Protein CASP antibody [EPR18806] (AB182216)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) cells labeling Protein CASP with ab182216 at 1/60 dilution (red) compared with a Rabbit IgG,monoclonal[EPR25A] -Isotype control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (Alexa Fluor^® 488) at 1/500 dilution was used as the secondary antibody.

Note : Cells were permeabilized with 90% methanol (in 1XPBS).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Protein CASP antibody [EPR18806] (AB182216)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) cells labeling Protein CASP with ab182216 at 1/100 dilution followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing specific but weak cytoplasmic staining on U-87 MG cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG (AlexaFluor^®594) preadsorbed (ab150120) at 1/1000 dilution (red).

The negative controls are as follows :

-ve control 1 : ab210528 at 1/100 dilution followed by ab150120 at 1/1000 dilution.

-ve control 2 : ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-Protein CASP antibody [EPR18806] (AB182216)

Protein CASP was immunoprecipitated from 1mg of HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate with ab182216 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab182216 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : HepG2 whole cell lysate, 10μg (Input).

Lane 2 : ab182216 IP in HepG2 whole cell lysate.

Lane 3 : Rabbit IgG, [EPR25A] - Isotype Control (ab172730) instead of ab182216 in HepG2 whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 3 seconds.

All lanes:

Immunoprecipitation - Anti-Protein CASP antibody [EPR18806] (ab182216)

Predicted band size: 77 kDa

false

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Protein CASP antibody [EPR18806] (AB182216)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling Protein CASP with ab182216 at 1/100 dilution followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing specific but weak cytoplasmic staining on NIH/3T3 cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG (AlexaFluor^®594) preadsorbed (ab150120) at 1/1000 dilution (red).

The negative controls are as follows :

-ve control 1 : ab210528 at 1/100 dilution followed by ab150120 at 1/1000 dilution.

-ve control 2 : ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Protein CASP antibody [EPR18806] (AB182216)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling Protein CASP with ab182216 at 1/120 dilution (red) compared with a Rabbit IgG,monoclonal [EPR25A] - Isotype control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (Alexa Fluor^® 488) at 1/500 dilution was used as the secondary antibody.

• WB

Supplier Data

Western blot - Anti-Protein CASP antibody [EPR18806] (AB182216)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane1,2,3 and 4 : 3 minutes; Lane 5,6 and 7 : 30 seconds.

All lanes:

Western blot - Anti-Protein CASP antibody [EPR18806] (ab182216) at 1/2000 dilution

Lane 1:

Rat testis lysate at 20 µg

Lane 2:

Rat cerebral cortex lysate at 20 µg

Lane 3:

Mouse cerebral cortex lysate at 20 µg

Lane 4:

Neuro-2a (Mouse neuroblastoma cell line) whole cell lysate at 20 µg

Lane 5:

Mouse testis lysate at 20 µg

Lane 6:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 7:

HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 77 kDa

Observed band size: 77 kDa

false

• WB

Supplier Data

Western blot - Anti-Protein CASP antibody [EPR18806] (AB182216)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1 and 2 : 30 seconds; Lane 3 : 3 minutes.

All lanes:

Western blot - Anti-Protein CASP antibody [EPR18806] (ab182216) at 1/2000 dilution

Lane 1:

HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 10 µg

Lane 2:

SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate at 10 µg

Lane 3:

U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 77 kDa

Observed band size: 77 kDa

false

• WB

Supplier Data

Western blot - Anti-Protein CASP antibody [EPR18806] (AB182216)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Protein CASP antibody [EPR18806] (ab182216) at 1/2000 dilution

Lane 1:

Human fetal liver lysate at 10 µg

Lane 2:

Human fetal heart lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/100000 dilution

Predicted band size: 77 kDa

Observed band size: 77 kDa

false

Exposure time: 30s

• WB

Supplier Data

Western blot - Anti-Protein CASP antibody [EPR18806] (AB182216)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1-6 : 30 seconds; Lane 7,8,9 and 10 : 8 seconds.

All lanes:

Western blot - Anti-Protein CASP antibody [EPR18806] (ab182216) at 1/2000 dilution

Lane 1:

Mouse brain lysate at 10 µg

Lane 2:

Mouse kidney lysate at 10 µg

Lane 3:

Mouse spleen lysate at 10 µg

Lane 4:

Rat brain lysate at 10 µg

Lane 5:

Rat heart lysate at 10 µg

Lane 6:

Rat spleen lysate at 10 µg

Lane 7:

C6 (Rat glial tumor cell line) whole cell lysate at 10 µg

Lane 8:

RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

Lane 9:

PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg

Lane 10:

NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 77 kDa,85 kDa

Observed band size: 77 kDa

false