Anti-Cytosolic Phospholipase A2 antibody (ab73406)
Key features and details
- Rabbit polyclonal to Cytosolic Phospholipase A2
- Suitable for: IHC-P, WB, ICC/IF
- Knockout validated
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-Cytosolic Phospholipase A2 antibody
See all Cytosolic Phospholipase A2 primary antibodies -
Description
Rabbit polyclonal to Cytosolic Phospholipase A2 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanIHC-P HumanWB Human
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Immunogen
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Positive control
- This antibody gave a positive signal in HeLa Whole Cell Lysate - Bleomycin Treated (40U/ml)
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General notes
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Images
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Western blot - Anti-Cytosolic Phospholipase A2 antibody (ab73406)
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: Cytosolic Phospholipase A2 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: A549 whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab73406 observed at 90 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab73406 was shown to specifically recognize Cytosolic Phospholipase A2 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when Cytosolic Phospholipase A2 knockout samples were examined. Wild-type and Cytosolic Phospholipase A2 knockout samples were subjected to SDS-PAGE. Ab73406 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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Immunocytochemistry/ Immunofluorescence - Anti-Cytosolic Phospholipase A2 antibody (ab73406)ICC/IF image of ab73406 stained HeLa cells. The cells were 4% PFA 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 (ab73406, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 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. This antibody also gave a positive result in 4% PFA fixed (10 min) Hek293, HepG2 and MCF7 cells at 1µg/ml, and in 100% methanol fixed (5 min) HeLa cells at 1µg/ml. -
Western blot - Anti-Cytosolic Phospholipase A2 antibody (ab73406)Anti-Cytosolic Phospholipase A2 antibody (ab73406) at 1 µg/ml + HeLa Whole Cell Lysate - Bleomycin Treated (40U/ml) at 10 µg
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 85 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?The 100kDa band is comparable to molecular weights seen with other commercially available antibodies to Cytosolic Phospholipase A2
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytosolic Phospholipase A2 antibody (ab73406)IHC image of Cytosolic Phospholipsae A2 staining in human colon carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab73406, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
