Anti-Osteopontin antibody (ab11503)

Thank you for your enquiry. ab11503 Osteopontin antibody has only been tested by ELISA, Neutralisation and western blotting. That is not to say that it will not work by immunhistochemistry; just that it is yet to be tested. Searching our on line catalogue I found an antibody against this target that has been shown to work by IHC-P; albeit with only a predicted reactivity against mouse based on sequence similarity of the immunogen. The antibody is: ab14175 Osteopontin antibody ELISA, IHC-P. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

The immunogen of antibody is from amino acid 170-183 of human Osteopontin http://www.uniprot.org/uniprot/P10451. The antibody should bind to full length protein as well the immunogen sequence containing fragment.

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The Isotype control ab37373 can be used with ab11503.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Both of these antibodies are IgG antibodies. Unfortunately, we have not determined the subclass(es) of IgG present but I would expect there to be a mix of subclasses with possibly one predominating but this could vary depending on the immune response of the rabbit for ab1790 and the mouse for ab11503.

Vielen Dank für Ihre Anfrage.

Es tut mir ausgesprochen leid, aber wir haben leider keinen Antikörper, der für Ihre Zwecke geeignet zu sein scheint. Haben Sie es schon auf der Biocompare Seite versucht? Diese bieten auch ein breites Sortiment von Antikörpern verschiedenster Hersteller an. Wenn Sie einen Antikörper suchen, der nur mit der Maus reagiert, sollte dieser gegen die AS in Position 120-130, 158-165, 180-190, 200-215 oder 225- 238 des Maus- Proteins gerichtet sein, da es in diesen Positionen die geringste Übereinstimmung gibt.

Vielen Dank für Ihre Geduld, und ich wünsche Ihnen viel Erfolg bei Ihrer Suche und Ihren Experimenten.

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I have received additional information from the lab regarding the procedure that was used.

1) How long should one incubate with the protein (for coating the wells)?
Answer: We coat the recombinant mouse osteopontin on the plates overnight at room temperature.

2) For how long should one incubate with the antibody (for neutralization step)?
Answer: We incubate the neutralizing antibody on the plate for 1 hour at 37°C prior to treating with cells. The cells are then added to the plate and incubated for 1 hour at 37°C.

I hope this additional information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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The lab sent me the following experimental details for neutralization:

Neutralization: this antibody has the ability to neutralize the bioactivity of recombinant mouse osteopontin. Immobilized recombinant mouse osteopontin (2 mg/ml) is incubated with various concentrations of the antibody (0.01 mg/ml to 40 mg/ml) for 1 hour at 37 °C in a 96 well plate. Following this preincubation period, 293 cells (1x10^6 cells/ml) (100 ml /well) is added. The total mixture is incubated at 37 °C for 45 minutes in a humidified CO2 incubator. At the end of the incubation, non-adherent cells are washed off. The cells attached to the wells are detected by measuring endogenous cellular lysosomal acid phosphatase activity.

The Neutralization Dose50 (ND50) for Anti-Osteopontin is 1-3 mg/ml in the presence of 2 mg/ml of recombinant mouse osteopontin using 293 cells.

The ND50 is the concentration of antibody required to yield one-half maximal inhibition of the cytokine activity on a responsive cell line, when the cytokine is present at a concentration just high enough to elicit a maximum response.

The exact concentration of antibody required to neutralize mouse osteopontin activity is dependent on the cytokine concentration, cell type, growth conditions, and the type of activity studied.

The lab is checking if additional information can be obtained from the originator. I will let you know if they do.

I hope this information is already helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Thank you for contacting us and thank you for submitting an Abreview for ab11503.

The testing discount codehas been activated when your Abreview was submitted. You can now use this code for ordering an Abcamprimary antibody of your choice.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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I am sorry you are experiencing problems to submit an Abreview. As you are using a cell model I have just chosen the option “cell” from Sample type, so the system allows you to continue completing the Abreview.

Any further information or clarification can be added to the Additional notes box at the bottom.

I hope this helps. Please do not hesitate to contact me back for further assistance.

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I am very pleased to hear you would like to accept our offer and test ab11503 in Rat species with Neutralising assays. This code will give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview 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: https://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.

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