Many of the early antibodies against ER-beta has questionable specificity.
Hi Joe,
Greetings from Poland. I hope everything is fine in Dublin.
In our study, we used the anti-ERβ Ab from Novacastra/Leica Wetzlar, Germany
http://www.breast-cancer-research.com/content/supplementary/bcr3691-s1.pdf
and the ZR-75-1 cell line:
http://www.breast-cancer-research.com/content/16/4/R79/figure/F5 (panel B)
We did not have decent negative controls (as this was just an addition to the main story), so I am not sure about the specificity. As ERbeta is quite ubiquitous, I think finding the negative control could be a challenge, but crucial if you are really concerned about the specificity.
Say hello the people in your lab from me.
Cheers,
Radek
Dear Michael,
In my hand, and after normalisation on HPRT1, HMBS and B2M, levels of ESR2 are much lower if compared to ESR2.
Please find below the ESR2 mRNA levels I measured by real-time RT-qPCR in our cell lines cultured in complete growth medium:
MDA-MB-415 0.0302
T47D 0.0281
SUM1315 0.0085
OCUB-F 0.0045
MDA-MB-157 0.0032
UACC812 0.0021
SKBR5 0.0017
SUM102PT 0.0014
MDA-MB-231 0.0014
EVSA-T 0.0014
SUM159 0.0012
DU4475 0.0011
MPE600 0.0011
SKBR3 0.0010
SUM229 0.0009
ZR75-1 0.0009
SUM44 0.0008
ZR75.30 0.0008
CAMA-1 0.0007
SKBR7 0.0006
MCF-7 0.0006
MDA-MB-436 0.0006
HCC1937 0.0005
SUM185 0.0005
MDA-MB-468 0.0005
MDA-MB-330 0.0004
MDA-MB-175 0.0004
SUM149 0.0003
BT549 0.0002
hs578t 0.0001
BT20 0.0001
BT483 0.0001
SUM52 0.0001
MDA-MB-453 0.0001
UACC893 0.0001
So MDA-MB-415 and T47D may serve as the most positive controls, and
hs578t, BT20, BT483, SUM52, MDA-MB-453 and UACC893 as negative controls.
All the best,
Anieta
Dear Michael,
What I've learnt while participating in this exactly topic duscussion during 37th Annual Breast Cancer Symposium in San Antonio (December 9-13, 2014) was that the mouse mammary tumor cell lines, D2A1 and MM51 both significantly express ER-alpha and ER-betha. Specifically D2A1 cells are dependent on estrogen for growth and have increased expression of aromatase. Additionally, D2A1 cells are very aggressive and highly metastatic - thus representing a good model of breast cancer progression. MM51 cells are Her2/neu positive and are an adequate model to study the crosstalk between growth factors ans ER signaling.
Best wishes,
Ilya
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