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On the basis of low sequence homology, ab183734 is predicted to show no or limited cross-reactivity to RFP, YFP, BFP, and CFP.
For the best results in IHC, please use ab183734 on Formalin/PFA-fixed paraffin-embedded sections with heat mediated antigen retrieval.
A trial size is available to purchase for this antibody.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
This product is a recombinant rabbit monoclonal antibody.
Our Abpromise guarantee covers the use of ab183734 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/10000 - 1/50000. Detects a band of approximately 27 kDa (predicted molecular weight: 27 kDa).|
|IHC-P||1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|Flow Cyt||Use at an assay dependent concentration.|
ab183734 staining GFP in GFP-transfected NIH3T3 cells. The cells were fixed with 4% formaldehyde (10min) and then blocked in 1% BSA / 0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab183734 at 1/500 dilution overnight at +4°C followed by incubation with ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488), for 1 hour, at 1μg/ml.
Under identical experimental conditions, when compared to the basal level of GFP expression in transfected NIH3T3 cells, the cells upon which ab183734 was applied gave a stronger signal in the 488 channel, indicating that ab183734 is binding to GFP and therefore eliciting signal amplification.
ab183734 was also applied to non-GFP-transfected NIH3T3 cells, which produced no positive staining, indicating specificity for GFP. Nuclear DNA was labelled with 1.43μM DAPI (blue).
Immunofluorescent analysis of 4% paraformaldehyde-fixed GFP transfected 293 cells labeling GFP with unpurified ab183734 at 1/500 dilution, followed by Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody at 1/200 dilution (green). Counterstained with Dapi (blue)
Blocking and dilution buffer: 5% NFDM /TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of GFP transgenic mouse pancreas tissue labelling GFP with purified ab183734 at a dilution of 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Flow Cytometry analysis of 293T (human embryonic kidney) transfected with human TNFRSF9 cells labeling GFP with purified ab183734 at 1/50 dilution(10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 647)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse pancreas tissue (negative control) labelling GFP with purified ab183734 at a dilution of 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Immunohistochemical analysis of paraffin-embedded GFP transgenic mouse colon tissue (left) and normal mouse colon tissue (right) labeling GFP with unpurified ab183734 at 1/250 dilution followed by prediluted HRP Polymer for Rabbit IgG. Counterstained with Hematoxylin.
Immunohistochemical analysis of paraffin-embedded GFP transgenic mouse liver tissue (left) and normal mouse liver tissue (right) labeling GFP with unpurified ab183734 at 1/250 dilution followed by prediluted HRP Polymer for Rabbit IgG. Counterstained with Hematoxylin.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"