Anti-Heparanase 1 antibody [EPR22365-230] (ab254254)
![Anti-Heparanase 1 antibody [EPR22365-230] (ab254254)](https://www.abcam.com/ps/products/254/ab254254/Images/ab254254-410187-anti-heparanase-1-antibody-epr22365-230-western-blot-human-knockout-lysate.jpg "Anti-Heparanase 1 antibody [EPR22365-230] (ab254254)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22365-230] to Heparanase 1
- Suitable for: WB
- Knockout validated
- Reacts with: Human
Overview
-
Product name
Anti-Heparanase 1 antibody [EPR22365-230]
See all Heparanase 1 primary antibodies -
Description
Rabbit monoclonal [EPR22365-230] to Heparanase 1 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species WB Human
-
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- WB: HeLa and Capan-1 whole cell lysates; Recombinant human active heparinase 1 protein (aa36-109 & 158-543).
-
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, PBS -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22365-230 -
Isotype
IgG -
Research areas
Images
-
Western blot - Anti-Heparanase 1 antibody [EPR22365-230] (ab254254)All lanes : Anti-Heparanase 1 antibody [EPR22365-230] (ab254254) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : HPSE knockout HeLa cell lysate
Lane 3 : Capan-1 cell lysate
Lane 4 : Human pancreas tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 61 kDa
Observed band size: 52,65 kDa why is the actual band size different from the predicted?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.
-
![Western blot - Anti-Heparanase 1 antibody [EPR22365-230] (ab254254)](https://www.abcam.com/ps/products/254/ab254254/Images/ab254254-2-anti-heparanase-antibody-epr22365-230-7-western-blot-recombinant-protein-human.jpg)
Western blot - Anti-Heparanase 1 antibody [EPR22365-230] (ab254254)Anti-Heparanase 1 antibody [EPR22365-230] (ab254254) at 1/1000 dilution + Recombinant human active heparinase 1 protein (aa36-109 & 158-543, expressed in CHo cells), 20 ng
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 61 kDa
Exposure time: 6 secondsBlocking and dilution buffer: 5% NFDM/TBST.
This blot was developed using a higher sensitivity ECL substrate.
-
![Western blot - Anti-Heparanase 1 antibody [EPR22365-230] (ab254254)](https://www.abcam.com/ps/products/254/ab254254/Images/ab254254-1-anti-heparanase-antibody-epr22365-230-7-western-blot-human.jpg)
Western blot - Anti-Heparanase 1 antibody [EPR22365-230] (ab254254)All lanes : Anti-Heparanase 1 antibody [EPR22365-230] (ab254254) at 1/1000 dilution
Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : Capan-1 (human pancreas adenocarcinoma epithelial cell line) whole cell lysate
Lane 3 : MCF7 (human breast adenocarcinoma cell line) whole cell lysate
Lane 4 : Human pancreas lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 61 kDa
Observed band size: 50,65 kDa why is the actual band size different from the predicted?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.
-
![Anti-Heparanase 1 antibody [EPR22365-230] (ab254254)](https://www.abcam.com/ps/products/254/ab254254/Images/ab254254-4-benefits-of-recombinant-antibodies.png)
Anti-Heparanase 1 antibody [EPR22365-230] (ab254254)
