Overview

  • 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

    Suitable for: IHC-P, WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat, Rabbit, Horse, Cow, Dog, Orangutan
  • Immunogen

    Synthetic peptide corresponding to Human Cytosolic Phospholipase A2 aa 700 to the C-terminus conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab85814, ab86259)

  • Positive control

    • This antibody gave a positive signal in HeLa Whole Cell Lysate - Bleomycin Treated (40U/ml)

Properties

Applications

Our Abpromise guarantee covers the use of ab73406 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 100 kDa (predicted molecular weight: 85 kDa).
ICC/IF Use a concentration of 1 µg/ml.

Target

Images

  • 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.

  • 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.
  • 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

  • IHC image of Cytosolic Phospholipsae A2 staining in humam 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.

References

ab73406 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Answer

Vielen Dank für Ihren Anruf.

Das Labor, das Anti-Phospholipase A2Antikoerper ab23705 herstellt, hat bestätigt, dass das Immunogen das Volllängenprotein sPLA2 (human Type IIA) mit der SwissProtID P14555.2 ist. Damit ist dieser Antikörper spezifisch für die Membran-assoziierte PLA2.

Die Antikörper ab124230 und ab73406 sind hingegen spezifisch für die cytosolische PLA2 mit der SwissProtID P47712.

Da diese beiden Formen sehr unterschiedlich sind, haben wir leider keinen Antikörper verfügbar, der beide Formen erkennt.

Ich hoffe, diese Informationen helfen Ihnen weiter. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

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Question
Answer

DISCOUNT CODE: *** Expiration date: May 3rd, 2012 I am very pleased to hear you would like to accept our offer and test ab73406 in mouse. This code will give you 1 freeprimary antibodybefore the expiration date. To redeem this offer, please submit an Abreview for ab73406 and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews. Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code. Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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Answer

Thank you for contacting us. We do not currently carry an antibody that meets your exact specifications, however you may be interested in ab73406. This antibody is raised in rabbit, and predicted to react with mouse. The immunogen sequence does not overlap either phosphorylation site, and it would therefore detect both the phosphorylated and non-phosphorylated forms of cPLA2. This antibody works for western blotting but has not been tested for use in a gel shift assay. I can offer a discount off a future purchase if you buy ab73406 now, test it in mouse and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free primary antibody. If you are interested in this offer, please follow these steps: 1. Reply to this e-mail to let me know that you would like to proceed and test ab73406 in mouse. I will then send a discount code. This code must be issued before purchasing ab73406 so please wait for my reply before ordering. 2. Purchase ab73406 either by phone, fax, or online (www.abcam.com). 3. Test it in mouse. 4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews. 5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any primary antibody ordered and the discount code is valid for 4 months after issue. We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab73406 turns out to be unsuitable for mouse, you will still receive the discount on your next purchase after your Abreview has been submitted. Please let me know if you have any questions about this offer and I would be happy to help you further. The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

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