Polyclonal antibody

About

Background: Researchers at Stanford have developed a polyclonal antibody against Myt1l, a protein involved in disease and direct reprogramming of neurons. Many research groups are using Myt1l in their studies, however current Myt1l antibodies are not optimal. To overcome this limitation, the inventors have created this polyclonal antibody against Myt1l. It is highly specific and well characterized and thus it will be in high demand. Stage of research: The antibody has been verified for specificity in Western blot (mouse and human), Immunofluorescence (mouse and human), Immunoprecipitation and Chromatin-Immunoprecipitation (mouse). Myt1l antibody design and characterization Legend a) Schematic of mouse MYT1 family members mmMYT1 (Q8CFC2), mmMyt1l (P97500), and mmST18 (A5LFV3) as well as human hsMyt1l homologue (Q9UL68). CCHC-type zinc fingers (ZF) as well as the following domains are highlighted in color: Nuclear localisation signals (NLS), aspartic acid/ glutamic acid-rich (Asp/Glu-rich), serine-rich (Ser-rich), MYT1, and coiled-coil domains. Also shown is the predicted antigenicity and the conservation between the proteins generated using EpiC and T-Coffee, respectively. Based on this a fragment of mmMyt1l spanning amino acids 171-420 was used as antigen for antibody induction in rabbits. Sequence identities among the antigen regions and the full-length proteins, as well as their molecular weights are shown (right). b-c) Western blot of; b) MEF cells upon induction of FLAG-tagged mmMyt1l and c) HEK293 cells upon transfection of FLAG-tagged mmMyt1, mmMyt1l, mmSt18, and untagged hsMyt1l using preimmune serum and antibodies against Myt1l, FLAG, and Tubulin. d) Microscopy images of HEK293 cells upon transfection of FLAG-tagged mmMyt1l followed by immunofluorescence using antibodies against FLAG (red), Myt1l (green), and DAPI staining (blue). Scale bar 10 µm. e-f) Western blot of; e) E13.5 embryonic mouse whole brain lysate and f) Myt1l ChIP eluates from (top) exogenous Myt1l in MEFs or (bottom) endogenous Myt1l from E13.5 embryonic mouse brain using preimmune serum and antibodies against Myt1l. g) Microscopy images of a section from adult mouse cortex upon immunofluorescence using antibodies against NeuN (red), Myt1l (green), and DAPI staining (blue). Scale bar 10 µm.   Applications:   Research tool- for use in: Western blots Immunofluorescence assays Immunoprecipitation assays Chromatin-Immunoprecipitation assays   Advantages:   Higher specificity as compared to existing antibodies- this antibody does not detect the other 2 similar family members by Western blot Well characterized Works in every application tested Can stain not only mouse, but also human Myt1l in brain slice   Publications:   Mall M, Kareta MS, Chanda S, Ahlenius H, Perotti N, Zhou B, Grieder SD, Ge X, Drake S, Euong Ang C, Walker BM, Vierbuchen T, Fuentes DR, Brennecke P, Nitta KR, Jolma A, Steinmetz LM, Taipale J, Südhof TC, Wernig M. Myt1l safeguards neuronal identity by actively repressing many non-neuronal fates. Nature. 2017 Apr 13;544(7649):245-249.