Products:Tags & Cell Markers >> Cell Type Markers >> Tumor Associated
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I was wondering if you could shed some light on the difference between this antibody and it's PE equivalent. The PE tagged version of this antibody does not have the B29.1 before the VU-ID9 tag, and I have had good success with the former. Would there be a difference in the two? |
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Thank you for contacting us. |
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Used in Flow Cytometry - datasheet use 1ug for 10tothe6 cells. He used 1ug to 10tothe 5 cells and got hardly any signal. Tried at 1:1 and got signal, but obviously used all ab at once for this. Mouse epithelium was tissue. |
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ANSWER: |
Thank you for your enquiry regarding Ab8666. We are very sorry to hear that you are having difficulty with this antibody. According to the datasheet, this antibody has been tested on human sample (breast, colon carcinoma and tonsil). The cross-reactivity with mouse protein has not been tested.
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Histogram plot of prostate tumour cells DU145 (green) over isotype control (red) stained with EpCam following resection.
This picture was kindly supplied as part of the review submitted by Phil Kearney.
ab8666 staining EpCAM in murine xenograft tumour derived from human DCIS cells by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). The sections were deparaffinized in xylene, and rehydrated in graded alcohols. Sections were then incubated at room temperature with ab8666 at 5 µg/ml. Slides were rinsed in borate buffer pH 8 and counterstained. EpCAM-FITC pseudo-colored green, 488 nm and DAPI psuedo-colored blue, 408 nm.
Image from Espina V et al, PLoS One. 2010 Apr 20;5(4):e10240, Fig 1.
ICC/IF image of ab8666 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab8666, 5µg/ml) overnight at +4ºC. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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