mouse gfap promoter sequence (InvivoGen)
Structured Review

Mouse Gfap Promoter Sequence, supplied by InvivoGen, used in various techniques. Bioz Stars score: 90/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/mouse gfap promoter sequence/product/InvivoGen
Average 90 stars, based on 10 article reviews
Images
1) Product Images from "Nkx2.1 regulates the generation of telencephalic astrocytes during embryonic development."
Article Title: Nkx2.1 regulates the generation of telencephalic astrocytes during embryonic development.
Journal: Scientific reports
doi: 10.1038/srep43093
Figure Legend Snippet: Figure 9. Binding of Nkx2.1 to conserved binding sequences in the promoters of various glial regulatory genes. Amplification of the putative Nkx2.1 binding sequences located in a 206 bp PCR product within the GFAP promoter (a) and in a 391 bp PCR fragment from the Lhx6 promoter (b) after chromatin immunoprecipitation (ChIP) with the Nkx2.1 antibody. Input DNA was added as the positive control as it contains the cross-linked sonicated genomic DNA taken before ChIP with the Nkx2.1 antibody; a strong signal was observed for all the promoter regions. No amplification of the Nkx2.1 core sequence (tcaag) located in two, 410 bp and 352 bp, PCR products from the Ngn2 promoter was detected (c). No signal was detected in the negative control where no antibody was added for ChIP nor when no DNA was added while performing the PCR (a–c). A very faint signal was detected in some samples immunoprecipitated with non-specific control IgG. These results suggest that Nkx2.1 binds in vivo to the promoter region of GFAP at putative Nkx2.1 binding sequences. The relative intensities of all the bands were calculated by assigning an arbitrary value of 1 to the input band. The quantifications are indicated below the gels. The Figure represents one of three independently performed assays. As a control, human embryonic kidney 293 (HEK293) cells were co-transfected with two reporter-constructs, namely, the pDRIVE-mGFAP-LacZ and the pCAG-IRES-Tomato plasmids (d–f). Cell nuclei were counterstained in blue with Hoechst (d). In the control, only 4.26% of Tomato+cells were LacZ+, n = 94. To test the binding of Nkx2.1 to the GFAP promoter, HEK293 cells were co-transfected with two reporter constructs, namely, the pDRIVE-mGFAP-LacZ and the pCAG-Nkx2.1-IRES-Tomato plasmids (g–i). Cell nuclei were counterstained in blue with Hoechst (g). Activation of the LacZ reporter was seen upon addition of the Nkx2.1 expression vector, thus confirming that Nkx2.1 activates the GFAP promoter, most probably by binding the sequence identified as Nkx2.1 consensus. Here, 98% of Tomato+cells were LacZ+, n = 50. Bar = 50 μm.
Techniques Used: Binding Assay, Amplification, Chromatin Immunoprecipitation, Positive Control, Sonication, Sequencing, Negative Control, Immunoprecipitation, Control, In Vivo, Transfection, Construct, Activation Assay, Expressing, Plasmid Preparation