Anti-RUNX3 antibody (ab68938)

Thanks for the recipe.
Here is a question. In the pre-experiment,when I used EDTA for antigen retrieval in microwave, the tissue sections all came off the slides. And when I used Sodium citrate, it seemed okay.Besides, when I used Na citrate first and then EDTA for retrieval, it also worked well.
Yet, I haven't proceededtothe IHCstaining process.
I'm writing to ask if I shall use Na citrate instead of EDTA for antigen retrieval? Or can I use Na citrate first and then EDTA in the retrieval process?

P.S.: My tissue section is IHC-P from embryonic mouse mandibular condyle, I didn't use EDTA for decalcification before embedding, cause it is cartilage at that moment. Primary antibodies are ab61753, ab102711, and ab68938.

ANSWER:

Thank you for your reply.


Regarding the EDTA retrieval, it is possible that this high pH solution is causing the tissues to fall off in which case, citrate buffer may be a suitable replacement. We have also found that buffer made in deionized water can cause tissues to fall off, but not in distilled water. Citrate treatment followed by EDTA buffer is not likely to improve your staining, but lead to over-retrieved samples and high background.

For ab61753, citrate buffer has been used successfully. We do not have this information for ab102711 or ab68938. Here is a recommended retrieval protocol:


1mM EDTA pH. 8.0

- Pour 1mM EDTA into the tank. Boil (3 min – high power)
- Place slides in the tank (5 min – med/low power)
- Remove tank from microwave. Top up with 1mM EDTA (5 min med/low power)
- Remove tank from microwave. Allow to cool at room temperature (20 min) & transfer to 1x TBS


I hope this information is helpful. Please do not hesitate to contact me if you have any additional questions.

Thank you for the reply.
Uh, What is the recipe of EDTA ph8.0 for antigen retrieval ? Do you mean 1mM EDTA adjusted to pH8.0 by using Tris? If I don't have Tris, can I use other acid/base for substitution ?

ANSWER:

Thank you for your reply.


Below please find the recipe for 1mM EDTA, adjusted to pH 8.0:


EDTA 0.37 g
Distilled water 1000 ml
Store at room temperature for 3 months.


Tris would not be used to pH a solution, but rather serves as a buffer. Please use HCL or NaOH to adjust pH.


I hope this information is helpful. Please do not hesitate to contact us if you have any additional questions.

Dear Sir/Madam Hi! I want to order the anti-Runx3 antibody for testing the expression of Runx3 in tissue section. I am writing to ask if there is any difference between your product of ab49117 and ab68938. BTW, my sample is IHC-P from mouse.

ANSWER:

Thank you for your inquiry.

For ab49117, we would guarantee this antibody to work in IHC-P in mouse, however we have not tested this in house. We know that IHC-P has been tested with human samples in this reference:

Nakanishi Y et al. Loss of runt-related transcription factor 3 expression leads hepatocellular carcinoma cells to escape apoptosis. BMC Cancer 11:3 (2011). WB, IHC-P; Human. PubMed: 21205319

For ab68938, we have more testing information with this antibody in IHC-P with mouse which you can see online.

http://www.abcam.com/RUNX3-antibody-ab68938.html

We would guarantee either antibody to work in IHC-P with mouse samples, but we do have more information about ab68938 in the way you would like to use the antibody.

I hope this information helps. Please contact us with any other questions.