Anti-PDGF BB antibody (ab23914)

Western blot - PDGF BB antibody (ab23914)

All lanes : Anti-PDGF BB antibody (ab23914) at 1 µg/ml

Lane 1 : PDGF BB protein (Active) (ab9706)
Lane 2 : PDGF BB protein (Active) (ab9706) with PDGF BB peptide (ab24035) at 1 µg

Lysates/proteins at 0.01 µg per lane.

Secondary
Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution

Performed under reducing conditions.

Predicted band size : 14-18 kDa
Observed band size : 14 kDa (why is the actual band size different from the predicted?)

ab23914 detects a band of ~ 14 kDa corresponding to the reduced form of recombinant PDGF BB (ab9706). The predicted molecular weight for PDGF BB is 27 kDa according to the Swissprot entry, however the reduced form is known to run at between 14-18 kDa. The 14 kDa band is specifically blocked by the addition of the immunizing peptide, ab 24035.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - PDGF BB antibody (ab23914)

Image courtesy of Human Protein Atlas

ab23914 staining PDGF BB in Human kidney (male). The paraffin embedded  tissue was incubated with ab23914 (1/500 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6. ab23914 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines.

Further results for this antibody can be found at www.proteinatlas.org

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - PDGF BB antibody (ab23914)

IHC image of PDGF BB staining in huan heart formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab23914, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

Western blot

Anti-PDGF BB antibody (ab23914) at 1 µg/ml + PDGF BB protein (His tag) (ab73231) at 0.01 µg

Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed (ab97080) at 1/5000 dilution
developed using the ECL technique

Performed under reducing conditions.

Exposure time : 1 minute