Anti-TMPRSS2 antibody [EPR3862]

4 product images

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  Western blot - Anti-TMPRSS2 antibody [EPR3862] (ab109131)

  False colour image of Western blot: Anti-TMPRSS2 antibody [EPR3862] staining at 1/2000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red.
  

  In Western blot, ab109131 was shown to bind specifically to TMPRSS2. A band was observed at 55, 25 kDa in wild-type LNCaP and at 25 kDa in Caco-2 cell lysates with no signal observed at this size in TMPRSS2 knockout LNCaP cell line Human TMPRSS2 knockout LNCaP cell line ab273745 (knockout LNCaP cell lysate Human TMPRSS2 knockout LNCaP cell lysate ab275499) and TMPRSS2 knockout Caco-2 cell line Human TMPRSS2 knockout Caco-2 cell line ab273737 (knockout Caco-2 cell lysate ab277340).

  To generate this image, wild-type and TMPRSS2 knockout cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.

  All lanes: Western blot - Anti-TMPRSS2 antibody [EPR3862] (ab109131) at 1/2000 dilution

  Lane 1: Wild-type LNCaP cell lysate at 20 µg

  Lane 2: TMPRSS2 knockout LNCaP cell lysate at 20 µg

  Lane 2: Western blot - Human TMPRSS2 knockout LNCaP cell line (Human TMPRSS2 knockout LNCaP cell line ab273745)

  Lane 3: Wild-type Caco-2 cell lysate at 20 µg

  Lane 4: TMPRSS2 knockout Caco-2 cell lysate at 20 µg

  Lane 5: PC-3 cell lysate at 20 µg

  Lane 6: DU 145 cell lysate at 20 µg

  Lane 7: Human Prostate cell lysate at 20 µg

  Lane 8: Human Colon cell lysate at 20 µg

  Lane 9: Human Lung cell lysate at 20 µg

  Performed under reducing conditions.

  Predicted band size: 54 kDa

  Observed band size: 25 kDa, 55 kDa

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMPRSS2 antibody [EPR3862] (ab109131)

  ab109131 staining TMPRSS2 in Human prostate adenocarcinoma tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed and paraffin-embedded, antigen retrieval was by heat mediation in Tris/EDTA buffer pH9. Samples were incubated with primary antibody (1/1000). An undiluted HRP-conjugated mouse anti-rabbit IgG was used as the secondary antibody. Tissue counterstained with Hematoxylin. PBS was used in the negative control rather than the Primary antibody.

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  Western blot - Anti-TMPRSS2 antibody [EPR3862] (ab109131)

  Blocking buffer and concentration: 5% NFDM/TBST
  Diluting buffer and concentration: 5% NFDM /TBST
  

  54kDa represents full-length TMPRSS2 and 25kDa is the cleaved form. The molecular weights observed are consistent with what have been described in the literature (PMID: 11245484).

  All lanes: Western blot - Anti-TMPRSS2 antibody [EPR3862] (ab109131) at 1/1000 dilution

  All lanes: LNCaP (Human prostate carcinoma epithelial cell) whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

  Predicted band size: 54 kDa

  Observed band size: 25 kDa, 54 kDa

  Exposure time: 7s

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMPRSS2 antibody [EPR3862] (ab109131)

  Immunohistochemical analysis of paraffin-embedded Human prostatic adenocarcinoma using ab109131, unpurified, at a dilution of 1/500.
  

  Perform heat mediated antigen retrieval before commencing with IHC staining protocol.