Anti-Collagen X antibody (ab58632)

Product code: 58632
Inquiry: Dear Support Team, Do you have any information if this collagen X antibody reacts with sheep cartilage? Thank you very much! Best regards, Andreja

ANSWER:

Thank you for contacting us and for your interest in our products.

I am sorry to confirm that as far as we are aware, ab58632 has never been tested with samples from sheep. All tested and guaranteedspecies cross-reactivity information is stated on our datasheets, and these are updated as soon as any new information is brought to our attention.

If you would like to test the antibody in this untested species, please contact our Scientific Support team by replying to this message prior to the purchase as you may be eligible for our testing discount program.

Otherwise, we like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates.

To find out more about our Abreview system, please see the following link:

http://www.abcam.com/abreviews

I hope this information is helpful. Please do not hesitate to contact me for any further advice or information in this regard.

I bought the following product rabbit polyclonal (ab58632) however, the concentration of the antibody is not specified in the data sheet. I need the stock concentration in order to calculate and determine my assay concentration and dilution.

ANSWER:

Thank you for contacting us.

The antibody is whole, unpurified antiserum, and the antibody concentration is not determined. Dilutions for some common applications are suggested on the datasheet. The appropriate isotype control for this antibody is whole normal rabbit serum, such as ab7487, diluted by the same factor as ab58632.

Click here (or use the following: http://www.abcam.com/index.html?datasheet=7487).

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

Dear technical team, Our customer would like to purchase 5 antibodies from abcam and wants to check control tissue. Pleaselet me knowabcam's product information. ab66025 ab102711 ab3773 ab2426 ab58632 Best regards,

ANSWER:

Thank you for your inquiry. Please find a list of possible positive controls below: ab2426: skin, lymph nodes Click here (or use the following: http://www.abcam.com/index.html?datasheet=2426). ab3773: cartilage samples Click here (or use the following: http://www.abcam.com/index.html?datasheet=3773). ab58632: Type X collagen is a product of hyperthrophic chondrotocytes and has been localized to presumptive mineralization zones of hyaline cartilage. Click here (or use the following: http://www.abcam.com/index.html?datasheet=58632). ab66025: The recommended positive control tissues are rat head tissue, rat embryo tissue, human osteosarcoma tissue, human glioma tissue, human intestinal cancer tissue. Click here (or use the following: http://www.abcam.com/index.html?datasheet=66025). ab102711: Specifically expressed in osteoblasts Click here (or use the following: http://www.abcam.com/index.html?datasheet=102711). I hope this information is helpful for your customer. I wish them good luck with their experiments.

What antigen retrieval method was used in IHC on paraffin-embedded tissue? What dilution do you recommend?

ANSWER:

Thank you for your call yesterday and for your patience while I have been in touch with the lab.

For ab58632, we recommend using the antibody at 1:50-1:100 and incubating overnight. The sections were treated with 0.25% trypsin in 1 mM EDTA for 3 minutes at 37C to expose the epitope.

These alterations should improve the staining results with the antibody, however if the problem persists, I would be happy to send a different antibody to try or alternatively issue a credit or refund.

Please keep me updated about any new results, and I look forward to hearing from you.

Thanks for your reply

Will the secondary antibody (e.g. Goat anti-Rabbit-IgG-HRP conjugated) creates non-specific signal when applied to a rabbit paraffin section?

Yours

ANSWER:

Thank you for your response.

It is always worth running a "no primary" (only secondary antibody) control to see/check if the secondary antibody alone may cause non-specific signal. In order to reduce the any potential non-specific signal, I would advise the following steps:

1) block the non-specific binding sites with 5% BSA (in TBS-T) for or with 10 % goat serum (host species of the secondary antibody) 1 hr at RT,

2) block endogenous peroxidase activity with H2O2 ,

You can find some more information on blocking at this site: http://www.abcam.com/index.html?pageconfig=resource&rid=11494

I hope this helps and if I can assist further, please do not hesitate to contact me.