Antibodies, Proteins, Kits and Reagents for Life Science

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Anti-Protein CASP antibody [EPR18806] (ab182216)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR18806] to Protein CASP
Suitable for: WB, Flow Cyt, IP, ICC/IF
Reacts with: Mouse, Rat, Human

Product name

Anti-Protein CASP antibody [EPR18806]
See all Protein CASP primary antibodies
Description

Rabbit monoclonal [EPR18806] to Protein CASP
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Mouse Human
ICC/IF	Mouse Human
IP	Human
WB	Mouse Rat Human

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human fetal liver and fetal heart lysates; HEK-293, SH-SY5Y, U-87 MG, Neuro-2a, HeLa, HepG2, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Rat testis and cerebral cortex; Mouse testis and cerebral cortex lysates; Mouse brain, kidney and spleen lysates; Rat brain, heart and spleen lysates. ICC/IF: NIH/3T3 and U-87 MG cells. Flow Cyt: U-87 MG and NIH/3T3 cells; IP: HepG2 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.

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, 0.05% BSA, 40% Glycerol (glycerin, glycerine)
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR18806
Isotype

IgG
Research areas

Western blot - Anti-Protein CASP antibody [EPR18806] (ab182216)

All lanes : Anti-Protein CASP antibody [EPR18806] (ab182216) at 1/2000 dilution

Lane 1 : Human fetal liver lysate
Lane 2 : Human fetal heart lysate

Lysates/proteins at 10 µg per lane.

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

Exposure time: 30 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-Protein CASP antibody [EPR18806] (ab182216)

All lanes : Anti-Protein CASP antibody [EPR18806] (ab182216) at 1/2000 dilution

Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 2 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate
Lane 3 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 77 kDa
Observed band size: 77 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1 and 2: 30 seconds; Lane 3: 3 minutes.
Western blot - Anti-Protein CASP antibody [EPR18806] (ab182216)

All lanes : Anti-Protein CASP antibody [EPR18806] (ab182216) at 1/2000 dilution

Lane 1 : Rat testis lysate
Lane 2 : Rat cerebral cortex lysate
Lane 3 : Mouse cerebral cortex lysate
Lane 4 : Neuro-2a (Mouse neuroblastoma cell line) whole cell lysate
Lane 5 : Mouse testis lysate
Lane 6 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 7 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 77 kDa
Observed band size: 77 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane1,2,3 and 4: 3 minutes; Lane 5,6 and 7: 30 seconds.
Western blot - Anti-Protein CASP antibody [EPR18806] (ab182216)

All lanes : Anti-Protein CASP antibody [EPR18806] (ab182216) at 1/2000 dilution

Lane 1 : Mouse brain lysate
Lane 2 : Mouse kidney lysate
Lane 3 : Mouse spleen lysate
Lane 4 : Rat brain lysate
Lane 5 : Rat heart lysate
Lane 6 : Rat spleen lysate
Lane 7 : C6 (Rat glial tumor cell line) whole cell lysate
Lane 8 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 9 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 10 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 77 kDa
Observed band size: 77 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1-6: 30 seconds; Lane 7,8,9 and 10: 8 seconds.
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.
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.
Flow Cytometry - Anti-Protein CASP antibody [EPR18806] (ab182216)

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.
Flow Cytometry - Anti-Protein CASP antibody [EPR18806] (ab182216)

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

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