Anti-PDK4 antibody [1C2BG5] (ab110336)
Key features and details
- Mouse monoclonal [1C2BG5] to PDK4
- Suitable for: Flow Cyt, WB, ICC/IF
- Knockout validated
- Reacts with: Human
- Isotype: IgG2a
Overview
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Product name
Anti-PDK4 antibody [1C2BG5]
See all PDK4 primary antibodies -
Description
Mouse monoclonal [1C2BG5] to PDK4 -
Host species
Mouse -
Specificity
This antibody does not cross-react with other isoforms of Human PDK (i.e. PDK1, 2 or 3). -
Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt HumanICC/IF HumanWB Human -
Immunogen
This information is considered to be commercially sensitive.
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Positive control
- WB: HeLa and HepG2 (ab7900) cell lysates and human heart tissue lysate. ICC/IF: HepG2 cells. Flow Cyt: HepG2 cells.
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General notes
This antibody clone is manufactured by Abcam.
Product was previously marketed under the MitoSciences sub-brand.
If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
Preservative: 0.02% Sodium azide
Constituent: HEPES buffered saline -
Concentration information loading...
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Purity
Proprietary Purification -
Purification notes
Produced in vitro using hybridomas grown in serum-free medium, and then purified by biochemical fractionation. Purity >95% by SDS-PAGE. -
Clonality
Monoclonal -
Clone number
1C2BG5 -
Isotype
IgG2a -
Light chain type
kappa -
Research areas
Images
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All lanes : Anti-PDK4 antibody [1C2BG5] (ab110336) at 1/500 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : PDK4 knockout HeLa cell lysate
Lane 3 : Human heart tissue lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 46 kDa
Observed band size: 48 kDa why is the actual band size different from the predicted?Lanes 1-3: Merged signal (red and green). Green - ab110336 observed at 48 kDa. Red - loading control, ab129002 observed at 124 kDa.
ab110336 Anti-PDK4 antibody [1C2BG5] was shown to specifically react with PDK4 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261805 (knockout cell lysate ab257217) was used. Wild-type and PDK4 knockout samples were subjected to SDS-PAGE. ab110336 and Anti-Vinculin antibody [EPR8185] - Loading Control (ab129002) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 1000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
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ICC/IF image of ab110336 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab110336, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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Overlay histogram showing HepG2 cells stained with ab110336 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab110336, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
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All lanes : Anti-PDK4 antibody [1C2BG5] (ab110336) at 1 µg/ml
Lane 1 : HepG2 whole cell lysate at 10 µg
Lane 2 : Human heart tissue at 10 µg
Lane 3 : Recombinant Human PDK1 at 0.01 µg
Lane 4 : Recombinant Human PDK2 at 0.01 µg
Lane 5 : Recombinant Human PDK3 at 0.01 µg
Lane 6 : Recombinant Human PDK4 at 0.01 µg
Predicted band size: 46 kDa