Anti-Vimentin antibody [RV202] - Neural Stem Cell Marker (ab8978)

  Thank you so much for helping me find primary (and secondary) antibodies for IHC on mouse neural progenitors down the lineage of cortex verses striatum. Attached is the paper (2012) that mentions cortex being identified with NCAM (Figure 4 i,j page 481) and Figure 1 says cortical induction efficiency was measured by Pax6, Vimentin, Otx1 and Otx2 expression (Figure 1c, page 478).   The second paper I attached (2008) is to help us figure out a good primary/secondary antibody detection of striatal differentiation. It says “Terminal striatal differentiation can be established by the detection ofMAP-2/Ki67[1] postmitotic neurons expressing key striatal markers, such as GABA, GAD67 (GAD1), DARPP32, ARPP21, calbindin, or calretinin.”   If you can find anything more efficient and specific for cortex vs. striatum it would be wonderful! Please contact me for any reason and thank you for your time and energy,

ANSWER:

Thank you for contacting us.

While there a many targets which express differentially in distinct layers of the neocortex many of these are expressed in the striatum as well. I found these papers to have a number of good targets which seem specific, such as Pax6 or, for striatal markers, DARPP32, CalB or GAD67. However a search through the Allen Mouse Brain Atlas expression data shows similar gene expression of all these markers in adult striatum and cortex. I would emphasize that this is not vetted data.

While any single or combination of these markers may well yield the information which you will be looking for, I feel that the best choices may be to try a few of these targets presented within this literature as you would have reference material to compare with.

Further review of our catalog leads me to suggest use of a DARPP32 antibody such as ab40801 alongside use of a Vementin antibody such as ab8978. Alternatively use of PAX6 ab5790 alongside GAD67, ab26116, may yield good results.

Each of these pairs will work in double staining as they are comprised of a mouse and a rabbit derived antibody. Further each product has been used in numerous publications and is well characterized by these, Abreviews and scientific support on the datasheet. Each is guaranteed to work in IHC-P as well as a number of other applications and would therefore be covered by our Abpromise guarantee:http://www.abcam.com/abpromise.

I have added links to each of these products below for your review:

ab40801;http://www.abcam.com/DARPP32-antibody-EP720Y-ab40801.html

ab8978;http://www.abcam.com/Vimentin-antibody-RV202-Neural-Stem-Cell-Marker-ab8978.html

ab5790;http://www.abcam.com/PAX6-antibody-Stem-Cell-Marker-ab5790.html

ab26116;http://www.abcam.com/GAD67-antibody-K-87-Neuronal-Marker-ab26116.html



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

Use our products? Submit an Abreview. Earn rewards!
http://www.abcam.com/abreviews

I have a question aboutab8978.
I found that it is mouse monoclonal and was tested in mouse sample on the data sheet.
I thinks that it is theoretically impossible because this antibody have to use anti-mouse IgGas secondary antibody.
So it can show high backgroundby non-specific bindingto endogenous mouse IgG especially in case ofIHC.
I would like to know how it is possible.
Please inform me.

ANSWER:

Thank you for contacting us.

Indeed, a mouse-on mouse staining can be quite tricky; however, it is not impossible. The background will depend on the fixation method ( perfusion fixed tissue vs. normally fixed tissue), on the tissue itself ( eg. kidneys vs. bone) and the experimental settings.


One way to make sure to decrease or to completelyerase the background is to block the endogenous mouse IgG with a non- conjugated secondary antibody before you incubate with your target antibody. I would suggest to use a Fab or F(ab)2 fragment antibody in this case as they will bound in a higher density than whole IgG antibodies.

You can find the aforementioned antibodies here:


http://www.abcam.com/index.html?datasheet=98754 (or use the following: http://www.abcam.com/index.html?datasheet=98754).

http://www.abcam.com/index.html?datasheet=6668 (or use the following: http://www.abcam.com/index.html?datasheet=6668).



Another way is to use our hassle- free Mouse on Mouse (M.O.M) Polymer IHC Kit, which contains a special rodent block solution:

http://www.abcam.com/index.html?datasheet=127055 (or use the following: http://www.abcam.com/index.html?datasheet=127055).


In addition, we also received an Abreview (17 July 2009) from one of our customers who rated this antibody highly in IHC. You can contact the author via our website if you like.



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

Use our products? Submit an Abreview. Earn rewards!
http://www.abcam.com/abreviews

LOT NUMBER GR57195-1 ORDER NUMBER 1015545 after centrifuge only 70ul has been found. Can you please arrange another vial ?

ANSWER:

This is to let you know that I have confirmed and agreed on placing a new order for your customer - for one vial of ab8978 as a free of charge replacement. I hope the second vial will work as it is expected, and please do let me know how you are getting on with this product.
Dear Colleague,
Please add 1 vial of ab8978 to the next shipment for Allied.
Original CCE ID: 3529205
Original Order: 1015545
Problem is: shortage 30 ul is missing (out of 100 ul)
Many thanks.

Our customer is going to test WB with rat sample and would like to know immunogen information, detection location.

ANSWER:

The immunogen was Cytoskeletal vimentin extract of calf lens and thus we do not know more exact immunogen region nor has the exact epitope been mapped.

we have used your antibody against vimentin, ab8978, for immunohistochemistry on paraffin-embedded formalin-fixed material. It seems the antibody works well on human intestinal samples. However, with mouse intestinal samples, we did not get a good signal. Can you provide some advice as how to proceed? What is you experience with immunohistochemistry on paraffin-embedded formalin-fixed with this antibody, in particular using mouse samples?

ANSWER:

Thank you for contacting us for technical support with ab8978. I'm glad to hear that this antibody is working well for you in human samples. I think the problem you are experiencing with this antibody in mouse tissue is due to the fact that when you add a secondary anti-mouse antibody this will bind everywhere on your mouse tissue; therefore you do not get specific signal. It is not recommended to use an antibody raised in the same species as your samples, for IHC.

I would recommend to try a primary antibody not raised in mouse for your mouse IHC, such as

ab7783 (raised in rabbit)

http://www.abcam.com/index.html?datasheet=7783

and ab39376 (raised in chicken)

http://www.abcam.com/index.html?datasheet=39376

Both antibodies work in mouse in IHC on paraffin sections.

I would like to encourage you to submit an Abreview on ab8978 via the online product datasheet. We always encourage customers to send their results back to us, whether positive or negative, and make all product information available to researchers.

To find out more about our Abreview system, please see the following URL: http://www.abcam.com/abreviews

I hope this information helps, please do not hesitate to contact us if you need any more advice or information.