Anti-Heparanase 1 antibody [EPR22365-230] - BSA and Azide free

• WB

Supplier Data

Western blot - Anti-Heparanase 1 antibody [EPR22365-230] - BSA and Azide free (AB255285)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure times.

Lane 1 : 3 minutes; Lanes 2 -4 : 78 seconds.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 16458254, 11406531).

Negative control : Human pancreas, MCF7 (PMID : 11406531, 10395325).

This blot was developed using a higher sensitivity ECL substrate.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254254).

All lanes:

Western blot - Anti-Heparanase 1 antibody [EPR22365-230] - BSA and Azide free (ab255285) at 1/1000 dilution

Lane 1:

HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

Capan-1 (human pancreas adenocarcinoma epithelial cell line) whole cell lysate at 20 µg

Lane 3:

MCF7 (human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 4:

Human pancreas lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 61 kDa

Observed band size: 50 kDa,65 kDa

false

• WB

Lab

Western blot - Anti-Heparanase 1 antibody [EPR22365-230] - BSA and Azide free (AB255285)

This data was developed using ab254254, the same antibody clone in a different buffer formulation.

Lanes 1-4 : Merged signal (red and green). Green - ab254254 observed at 52,65 kDa. Red - loading control ab8245 observed at 36 kDa.

ab254254 Anti-Heparanase 1 antibody [EPR22365-230] was shown to specifically react with Heparanase 1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265413 (knockout cell lysate ab258456) was used. Wild-type and Heparanase 1 knockout samples were subjected to SDS-PAGE. ab254254 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Heparanase 1 antibody [EPR22365-230] (<a href='/en-us/products/primary-antibodies/heparanase-1-antibody-epr22365-230-ab254254'>ab254254</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

HPSE knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human HPSE (Heparanase 1) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-hpse-heparanase-1-knockout-hela-cell-line-ab265413'>ab265413</a>)

Lane 3:

Capan-1 cell lysate at 20 µg

Lane 4:

Human pancreas tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 61 kDa

Observed band size: 52 kDa,65 kDa

false

• WB

Supplier Data

Western blot - Anti-Heparanase 1 antibody [EPR22365-230] - BSA and Azide free (AB255285)

Blocking and dilution buffer : 5% NFDM/TBST.

This blot was developed using a higher sensitivity ECL substrate.

All lanes:

Western blot - Anti-Heparanase 1 antibody [EPR22365-230] - BSA and Azide free (ab255285) at 1/1000 dilution

All lanes:

Recombinant human active heparinase 1 protein (aa36-109 & 158-543, expressed in CHo cells), 20 ng

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 61 kDa

false