Recombinant
RabMAb

Anti-Protein CASP antibody [EPR18806] (ab182216)

Overview

  • Product name
    Anti-Protein CASP antibody [EPR18806]
  • Description
    Rabbit monoclonal [EPR18806] to Protein CASP
  • Tested applications
    Suitable for: WB, Flow Cyt, IP, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human Protein CASP aa 400-650. The exact sequence is proprietary.
    Database link: Q13948

  • 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 rabbit monoclonal antibody.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

Properties

  • 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, 0.05% BSA, 40% Glycerol
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    EPR18806
  • Isotype
    IgG

Applications

Our Abpromise guarantee covers the use of ab182216 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/2000. Detects a band of approximately 77 kDa (predicted molecular weight: 77 kDa).
Flow Cyt 1/120.
IP 1/40.
ICC/IF 1/100.

 ab182216 binds weakly in ICC

Target

Images

  • 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
    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.

  • 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
    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.

  • 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
    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.

  • 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
    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.

  • 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.

  • 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 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 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).

  • 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 secondary antibody (HRP) (ab131366), was used as secondary antibody 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.

References

ab182216 has not yet been referenced specifically in any publications.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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