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Full length native protein from human erythrocytes.
Our Abpromise guarantee covers the use of ab9485 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IF||Use a concentration of 5 µg/ml.|
|ELISA||1/2500 - 1/5000.|
|WB||1/2500. Detects a band of approximately 40 kDa (predicted molecular weight: 37 kDa).
Some customers have experienced that milk significantly decreases the signal in WB compared to BSA. In-house we use BSA.
|ICC/IF||Use a concentration of 5 µg/ml.|
|Flow Cyt||Use at an assay dependent concentration.|
ab9485 staining GAPDH in HeLa cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab9485 at 5μg/ml and ab7291 at 1µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an AlexaFluor®488 Goat anti-Rabbit secondary (ab150081) at 2 μg/ml (shown in green) and AlexaFluor®594 Goat anti-Mouse secondary (ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.
Negative controls: 1– Rabbit primary antibody and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab9485 overnight at 4°C. Antibody binding was detected using ab175781 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab9485 overnight at 4°C. Antibody binding was detected using an anti-rabbit HRP secondary antibody, and visualised using ECL development solution ab133406.
Western blot image using the Optiblot Reducing Electrophoresis Kit - 10 x 10 cm (4-20%) (ab119220) with the
Prism Ultra Protein Ladder (ab116028) 5µl used. We recommend using our ECL substrate kit (ab65623) .
20ug of Lysate per lane and detection using ab9485 diluted to 1ug/ml.
Lane 1: Hela cell lysate
Lane 2: Jurkat cell lysate
Lane 3: A431 cell lysate
Lane 4: HEK293 cell lysate
Lane 5: HepG2 cell lysate.
This image is a courtesy of Anonymous Abreview
This picture was generously supplied by Dr Allison Robb of the MRC-LMB, Cambridge, UK.
This GAPDH antibody works well with an S. cerevisiae lysate in Western blotting.
A band can be seen down to a dilution of 1/5000.
A: 1:1000, 25 ug lysate
B: 1:1000, 50 ug lysate
C: 1:2000, 25 ug lysate
D: 1:2000, 50 ug lysate
E: 1:5000, 25 ug lysate
F: 1:5000, 50 ug lysate
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"