Anti-PEDF antibody

• WB

Supplier Data

Western blot - Anti-PEDF antibody (AB14993)

Chemiluminescence detection system.

All lanes:

Western blot - Anti-PEDF antibody (ab14993) at 1 µg/mL

All lanes:

Y79 (Human retinoblastoma cell line) cell lysate in RIPA buffer at 40 µg

Secondary

All lanes:

IgG (HRP) goat anti-rabbit

Predicted band size: 46 kDa

false

• WB

AbReview12162****

Western blot - Anti-PEDF antibody (AB14993)

Lanes 1-4 : Cartilage lysates from 4 osteoarthritic knee replacement donors (100 mg (wet weight) powdered cartilage/ 1 ml RIPA buffer, extracted for 24 hrs at 4°C)

Lane C : Positive control (overexpressing cell line)

Anticipated band at approx 50kDa detected. Specificity of antibody allowed relatively long exposure and quite high concentrations of primary and secondary antibodies to be used with negligible background. Good results were also obtained with unreduced samples.
The additional approx 250kDa band is likely to be due to PEDF crosslinked or bound to high molecular weight matrix proteins (collagen, aggrecan) which are present in cartilage lysates. Reduction using beta mercaptoethanol reduced some, but not all high MW staining and increased the intensity of the 50kDa band.

All lanes:

Western blot - Anti-PEDF antibody (ab14993)

Predicted band size: 46 kDa

true

Exposure time: 30s

Kindly submitted by Dr John Wardale.