SFPQ, rabbit pAb, Abcam, ab50935

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One User Review

  • Used for Western Blot and immunofluorescence detection of SFPQ protein.

    Experimental details:

    Western blot (HeLa cells) :
    -primary antibodies dilution: 1:2500-5000 of SFPQ antibody in 5%milk/PBS-T (incubation time: 1hr at RT or ON at 4 degrees). For ON incubation I have routinely used 1-5000 dilution.

    -secondary antibodies dilution: 1:10.000 of goat anti-rabbit/HRP antibodies in 5%milk/PBS-T (DAKO #P0448) (incubation time: 1hr at RT),

    -chemiluminescent substrate: SuperSignal West Femto (Thermo Scientific #34096)

    NB! Protein was detected between 100~130kDa band of PageRuler Prestained Protein Ladder (Thermo Scientific #26616). Predicted weight 76 kDa. Manufacturer claims that they detect protein 77 kDa. However, someone has submitted a review to the Abcam page and he/she also detects protein with larger size (~118 kDa). Importantly, this band is much weaker in the SFPQ siRNA treated cells (see Abcam review page for the results).

    Immunofluorescence (IF):

    Hela cells fixed with 4%PFA/PBS, 15 min at RT. For permeabilization I have used 70% EtOH.

    -SFPQ antibody dilution: 1:500-1000 in 1% BSA/PBS (incubation time: 1hr at RT). Signal predominately in the nucleus.

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Western blot
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