Anti-SCAP antibody (ab91323)

Overview

  • Product nameAnti-SCAP antibody
    See all SCAP primary antibodies
  • Description
    Rabbit polyclonal to SCAP
  • Specificityab91323 recognises isoform 4
  • Tested applicationsSuitable for: IP, WB, ELISAmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    KLH conjugated synthetic peptide selected from the center region of Human SCAP (NP_036367.2).

  • Positive control
    • K562 cell line lysate This antibody gave a positive result in IF in the following Methanol fixed cell line: HepG2

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab91323 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
IP
WB
ELISA
  • Application notesELISA: 1/1000.
    WB: 1/100 - 1/500. Predicted molecular weight: Isoform 1, 140 kDa; Isoform 4, 96 kDa.


    Not yet tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • Target

    • FunctionEscort protein required for cholesterol as well as lipid homeostasis. Regulates export of the SCAP/SREBF complex from the ER upon low cholesterol. Formation of a ternary complex with INSIG at high sterol concentrations leads to masking of an ER-export signal in SCAP and retention of the complex in the ER. Low sterol concentrations trigger release of INSIG, a conformational change in the SSC domain of SCAP, unmasking of the ER export signal, recruitment into COPII-coated vesicles, transport to the Golgi complex, proteolytic cleavage of SREBF in the Golgi, release of the transcription factor fragment of SREBF from the membrane, its import into the nucleus and up-regulation of LDLR, INSIG1 and the mevalonate pathway.
    • Sequence similaritiesBelongs to the WD repeat SCAP family.
      Contains 1 SSD (sterol-sensing) domain.
      Contains 7 WD repeats.
    • DomainCholesterol bound to SSC domain of SCAP or oxysterol bound to INSIG1/2 leads to masking of an ER export signal on SCAP possibly by moving the signal further away from the ER membrane.
    • Cellular localizationEndoplasmic reticulum membrane. Golgi apparatus membrane. Cytoplasmic vesicle > COPII-coated vesicle membrane. Moves from the endoplasmic reticulum to the Golgi in the absence of sterols.
    • Information by UniProt
    • Database links
    • Alternative names
      • KIAA0199 antibody
      • SCAP antibody
      • SCAP_HUMAN antibody
      • SREBF chaperone antibody
      • SREBF chaperone protein antibody
      • SREBP cleavage activating protein antibody
      • SREBP cleavage-activating protein antibody
      • Sterol regulatory element binding protein cleavage-activating protein antibody
      • Sterol regulatory element-binding protein cleavage-activating protein antibody
      see all

    Anti-SCAP antibody images

    • ab91323 stained HepG2 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab91323 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    • Anti-SCAP antibody (ab91323) at 1/1000 dilution + K562 whole cell lysate at 35 µg

      Secondary
      Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution

      Predicted band size : Isoform 1, 140 ; Isoform 4, 96 kDa

      Blocking/Dilution buffer: 5% NFDM/TBST.

    • SCAP was immunoprecipitated using 0.5mg K562 whole cell extract, 5µg of Rabbit polyclonal to SCAP and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).

      The antibody was incubated under agitation with Protein G beads for 10min, K562 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.

      Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab91323.

      Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).

      Bands: isofrom 1 - 140kDa, isofrom 4 - 96KDa; SCAP

    • Anti-SCAP antibody (ab91323) at 1/100 dilution + K562 whole cell lysate at 35 µg

      Predicted band size : Isoform 1, 140 ; Isoform 4, 96 kDa

    References for Anti-SCAP antibody (ab91323)

    ab91323 has not yet been referenced specifically in any publications.

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    Thank you for contacting us and your interest in our products.

    The immunogen sequence used to raise the anti-SCAP antibody (ab91323) is proprietary information. However, in order to aid your research I am able to share it with you. Please do...

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    Application Western blot
    Sample Mouse Tissue lysate - whole (Nerve lysate)
    Loading amount 40 µg
    Specification Nerve lysate
    Gel Running Conditions Reduced Denaturing
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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    Submitted Jul 20 2012

    DISCOUNT CODE: ***
    Expiration date: September 9, 2012

    This code will give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for ab125186 using mouse or rat samplesand include this cod...

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    I am sending a vial of ab86415 free of charge on the order ***. This antibody is in stock and should arrive on Monday. Please keep me updated about how this antibody works in your W...

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    DISCOUNT CODE: *** Expiration date: March 30th, 2012 I am very pleased to hear you would like to accept our offer and test ab91323 in mouse. This code will give you 1 free primary antibody before the expiration date. To redeem this offer, please submit...

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    Thank you very much for your interest in ab91323. To our knowledge, ab91323 has not been tested on mouse samples, but the immunogen sequence is 90% homologous with the mouse protein. Therefore, I can offer a discount off a future purchase if you buy ab...

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