Anti-PLVAP/PV-1 antibody [MECA-32] (ab27853)

Thank you for your patience.

For ab27853, it is as I had thought in my last email.The application was added based on publications showing that [MECA-32] has worked in IHC-Fr, e.g.

"The Adnectin CT-322 is a novel VEGF receptor 2 inhibitor that decreases tumor burden in an orthotopic mouse model of pancreatic cancer" by Dineen et al., BMC Cancer 2008, 8:352 (see attachment).

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

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry the product ab2350 did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number xxx with ab27853.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

As for the IHC-Fr protocols:


For ab27853, I will need to check with the supplying lab if they have any details on that. As far as I know, it was reported to them by other researchers that the antibody works in IHC-Fr. I will let you know what I find out.

I have attached our IHC-Fr protocols to help you further in the meantime.


For ab53444,we have the following protocol on file:

Indirect Immunostaining of Frozen Tissue Sections

For use with unconjugated monoclonal and polyclonal antibodies.
Please note that a certain level of technical skill and immunological knowledge is required for the successful design and implementation of these techniques - these are guidelines only and may need to be adjusted for particular applications.
Reagents
1. 0.3% H2O2‎, 70% methanol in TBS
1 ml of 30% H2O2‎ per 100 ml methanol/TBS.
2. TBS stock solution (10x)
NaCl, 87.66 g
Tris base, 60.55 g
Distilled water, 1 liter
Adjust pH to 7.4 using concentrated HCl.
Method
In most cases, we recommend that tissues are snap-frozen in liquid nitrogen, then prepared as 4-6 μm sections using a cryostat.
Allow sections to air dry for at least 1 hour.
Fix sections in dry acetone for 15 minutes. Allow to evaporate for 10 minutes.
Block endogenous peroxidase (if necessary) by immersing slides in 0.3% H2O2‎ in 70% methanol/TBS for 30 minutes. Wash once in TBS.
Incubate sections for 10 minutes in 10% normal serum from the species in which the secondary was raised. Tap excess serum off the slides before staining.
Incubate sections with primary antibody for at least 30 minutes at room temperature in a humid chamber, or overnight at 4°C. Wash 3 times in TBS.
Add enzyme-conjugated secondary antibody at the recommended dilution (see specific datasheet for details). Incubate for at least 30 minutes at room temperature. Wash 3 times in TBS.
Incubate with the appropriate substrate solution for the recommended period of time (it is suggested to useDAB substrate with HRP-conjugated antibodies, and Fast Red/Napthol AS-MX for Alkaline Phosphatase-conjugated antibodies). Wash once in water.
Counterstain with hematoxylin for 1-10 minutes. “Blue” with running water for 5 minutes. Then wash.
Mount in aqueous mounting medium, or alternatively dehydrate through a graded series of alcohols and xylene/solvent, and mount in synthetic mountant.

Notes
- Do not allow slides to dry out after the fixation step, as drying will result in damage to the tissue structure.
- Beware, certain substrates are soluble in alcohol – please refer to manufacturer's information for details.
- Appropriate controls should always be carried out. It may be useful to include a sample in which no primary antibody is used at all, in order to determine any non-specific binding of the secondary reagent to the target tissue.

In addition, you can also check the Abreviews section where 7 IHC-Fr reviews had been submitted by other customers:
https://www.abcam.com/CD68-antibody-FA-11-ab53444-reviews.html




I wish you the best of luck with your research.

Please let me know if there is anything else I can do for you.

Thank you for contacting us. While I am unaware of any references describingACVRL1 expression in tumors, the human protein atlas (linked below) does show that "distinct cytoplasmic staining with a granular pattern was observed in thyroid cancers, malignant lymphomas and in a few malignant melanomas, colorectal, gastric, pancreatic and renal cancers. Malignant cells in general were negative." http://www.proteinatlas.org/ENSG00000139567/cancer Additionally, because ACVRL1 is88%homologous betweenmouse and human, it will be difficult to find an antibody to that target which will react with mouse and not human epithelial cells. If you are looking to specifically detect mouse endothelial cells, you may be interested in the PLVAP antibody clone MECA-32 (ab27853), which is described in http://www.ncbi.nlm.nih.gov/pubmed/7626790?dopt=AbstractPlus. I hope this helps, please let me know if you need any additional information.