Overview

  • Product name
  • Description
    Goat polyclonal to CASP
  • Host species
    Goat
  • Tested applications
    Suitable for: WB, ELISAmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide corresponding to Human CASP aa 2-14 (N terminal).
    Sequence:

    SLQRLLQHSSNGN-C


    (Peptide available as ab22893)

  • Positive control
    • Jurkat lysate.
  • General notes
    GenBank Accession Number – NP_004279.

    Protein previously labeled as PSCDBP.

Properties

Applications

Our Abpromise guarantee covers the use of ab2247 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 Use a concentration of 1 - 3 µg/ml. Detects a band of approximately 40 kDa (predicted molecular weight: 41 kDa).Can be blocked with Human CASP peptide (ab22893).
ELISA 1/16000.

Target

  • Function
    By its binding to cytohesin-1 (CYTH1), it modifies activation of ARFs by CYTH1 and its precise function may be to sequester CYTH1 in the cytoplasm.
  • Tissue specificity
    Expressed in lymph nodes, thymus, spleen, lung, peripheral blood leukocytes and bone marrow.
  • Sequence similarities
    Contains 1 PDZ (DHR) domain.
  • Cellular localization
    Cytoplasm. Early endosome. Recruited from the cytosol to endosomes by SNX27.
  • Information by UniProt
  • Database links
  • Alternative names
    • B3-1 antibody
    • B31 antibody
    • CASP antibody
    • Cbp HE antibody
    • CYBR antibody
    • CYTHIP antibody
    • Cytip antibody
    • CYTIP_HUMAN antibody
    • Cytohesin 1 interacting protein antibody
    • Cytohesin binder and regulator antibody
    • Cytohesin-associated scaffolding protein antibody
    • Cytohesin-binding protein HE antibody
    • Cytohesin-interacting protein antibody
    • HE antibody
    • Pleckstrin Homology Sec7 and Coiled-Coil Domains Binding Protein antibody
    • Pleckstrin homology Sec7 and coiled-coil domains-binding protein antibody
    • Pleckstrin homology, Sec7 and coiled/coil domains, binding protein antibody
    see all

Images

  • ab2247 staining (1µg/ml) of Jurkat lysate (RIPA buffer, 35µg total protein per lane).  Primary incubated for 1 hour.  Detected by western blot using chemiluminescence. ab2247 staining (1 µg/ml) of MOLT4 cell lysate (RIPA buffer, 35µg total protein per lane). Primary incubated for 1 hour. Detected by western blot using chemiluminescence

References

This product has been referenced in:
  • O'Brien M  et al. Upregulation of PSCDBP, TLR2, TWIST1, FLJ35382, EDNRB, and RGS12 gene expression in human myometrium at labor. Reprod Sci 15:382-93 (2008). Read more (PubMed: 18497345) »
  • MacNeil AJ  et al. Sorting nexin 27 interacts with the Cytohesin associated scaffolding protein (CASP) in lymphocytes. Biochem Biophys Res Commun 359:848-53 (2007). IP ; Human . Read more (PubMed: 17577583) »
See all 2 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ELISA
Sample
Human Cell (AGS human gastric carcinoma cells)
Specification
AGS human gastric carcinoma cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Type
Sandwich (Detection)

Abcam user community

Verified customer

Submitted Jul 27 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (AGS Gastric Carcinoma cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
10 µg
Specification
AGS Gastric Carcinoma cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Jun 17 2010

Answer

Thank you very much for your enquiry and patience. Below is the Western blotting protocol that the originator used with ab2247. I hope this helps and please let me know if you need further assistance. - Lysis. Cell pellets were washed with ice-cold PBS. 1 ml of RIPA buffer was added per 1E8 cells and incubated on ice for 20 min, vortexing 2-3 times, briefly. The lysate was aliquotted into 1.5 ml microfuge tubes and centrifuged at 13,000 rpm for 5 min in a microfuge. The supernatant was transferred into clean tubes and its protein concentration was measured with BioRad protein assay. The concentration was then adjusted to 5 mg/ml with RIPA lysis buffer. An equal volume of 2 x SDS sample buffer was then added and the cell lysate was boiled for 5 minutes. Lysates were stored at -80C until use.(RIPA buffer = 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1 mM PMSF, 1 mM EDTA, 5 µg/ml Aprotinin, 5 µg/ml Leupeptin, 1% Triton X-100, 1% Sodium deoxycholate, 0.1% SDS). - SDS PAGE. Samples were run at 200V constant on a 12% acrylamide SDS-PAGE mini gel - using Biorad Mini-Protean 3 kit and protocols. Before loading samples had 5% (v/v) 2-ME added and were boiled for 3 minutes. - Transfer. We used a Biorad Mini Trans-Blot, constant 100 V for 1 hour. Transfer Buffer was 20 mM Tris pH 8.0, 150 mM Glycine, 10% Methanol. We transferred to Millipore PVDF membrane and stained with Ponceau Red to evaluate the transfer. - Staining. The membrane was blocked in 2.5% skimmed milk in TBS-T (TBS + 0.05% Tween-20) for 1 hr at room temperature with agitation. Primary antibody was incubated for 1 hr at room temperature with agitation. We used sigma secondary (Sigma anti-goat-HRP Product # A4174, use 1:3000) for 1 hr at room temperature with agitation. We washed with TBST three times after primary and secondary antibody, each wash lasting for 5-10 mins. ECL-plus (Amersham) was used rather than ECL, which is considerably more sensitive. Final detection was on autoradiography film.

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