Heating at 70°C for 5–10 min is also acceptable and may be preferable when studying multi-pass membrane proteins. H��W]o7|ׯ�K �Hy�a v�E��E!ɪ�V: �ݤ��3K�ɑ��h��0 ������. 0000022507 00000 n 0&6��s��8���#?�&�������N��� 0 wy endstream endobj 122 0 obj [/ICCBased 141 0 R] endobj 123 0 obj <> endobj 124 0 obj <> endobj 125 0 obj <> endobj 126 0 obj <>stream P"�lV���@�֘@Z����Ux�&;�(M``��\`,4���I��iR�k83�q6ݙP��e��Qٰ�)��!������+ʅ���:g����ò��u�S���x;@� ��o� endstream endobj 117 0 obj <>>> endobj 118 0 obj >/PageWidthList<0 612.0>>>>>>/Resources<>/ExtGState<>/Font<>/ProcSet[/PDF/Text]/XObject<>>>/Rotate 0/TrimBox[0.0 0.0 612.0 792.0]/Type/Page>> endobj 119 0 obj <> endobj 120 0 obj <> endobj 121 0 obj <>stream bc&7&��ufr��M�b�0t����rx!� ��8o�X�ϐOB4��i��N#n0�#�^F_��)����Q�8�x��1#�*ybatC�:Q��oa�e�K�����\&�J��[��}�m�uf���Nd ��C���%zm���׈����"Tα��n�ѫ���x�vx>���L��R��71xF��fp��? 0000010324 00000 n 0000007341 00000 n 2) Add methanol and mix. Loading buffer, running buffer, coomassie brilliant blue staining solution, and coomassie destaining solution are needed to be prepared for SDS-PAGE, while western blot transfer buffer (recipe here is for wet transfer) preparation is required for protein transfer. Cool to room temperature. 0000011772 00000 n s�-�3�3�3�3�3�3�-�-�-�-�-�M�v�ڕ55�55�55�kW�]����������s}�}�s���+sP��A��2E�س�A�9s0�`�7�� �F�����o�7�� �F�����o�7�� �ԩ� -��*Uu �,��d�[�&�q܋�����n��#l��.���~�����?ƾ>�ɝۮ���N��vYY����Go~����i��~��uߝ�l�t����6�w��n�S|���cǶ�����ó����7^c7�­V���T��q�v�F���^�Mz�N��4��_�!�j��&ކc�cw�ٲH��-�ҵ�b������J~/�_��k���;0�LMbΗ�l�9��\�$���\�=ك�,`�y�y%����t�p�t�p�t�p����:����A� p:����A� p:dCހ�� 7an�܄� rz 116 33 0000025156 00000 n 0000015261 00000 n ��:%��#F:�ĝ��?dJ��l1��i���~3?c��+�޹P���7P��v�����I�����>ZO���:GO����~�/rqy�>"g�S��Ǜ�{�0���o��1���?ob�6��!6E��^��_lJ��Mt:'y����q�;��K����N1ڶ.�W�������94ϐ�hN���F)P70���`C�'6`w�6�AY~�c0�:E-6"��:W�5[c^3�N*X� 8�(aoT��*T(*�� 0000008733 00000 n �?Ȳ endstream endobj 130 0 obj <> endobj 131 0 obj <>stream }���9�|>��k�y;ݞ���nCrż_��t�:�UwJY��k��7�VY�����ñ~�Ƭ\~��U�_V��ګ�ż��t������/�8_l�7�[�-4}l1M[�ҳ�G}��^B�����B-�����J f�#49=8=9=8��zmZ�+�� %PDF-1.5 %���� h�b```b``�c`e`�� ̀ �@16�GA3�Hp�o�`NcH0q`�m``��ۻuuT$2Pd�K�`��2'�Lb�84�|��Fش2�؏l�,9ZyU���f�'N=,���1�qBȡ:�yS��b&�U1�y��h �Y�zP �C�R~�B�ҥ1�l�V%v15(�`s�r����+��d`0q�q�[email protected]_��LJJJ��P� Tris-glycine native running buffer: 25 mM Tris base, 192 mM glycine, pH 8.3 Recipe for 10X buffer stock: Tris base 29 g Glycine 144 g Deionized water to 1,000 mL MOPS SDS running buffer: 50 mM MOPS, 50 mM Tris base, 0.1% SDS, 1 mM EDTA, pH 7.7 Recipe for 20X buffer stock: MOPS 104.6 g Tris base 60.6 g SDS 10 g EDTA 3.0 g Deionized water to 500 mL H�\��n�@���C����$���ɇ��z�0�����vQV"-��t}բ�ov�]�N.�5��>M��v͘.�u��;�S�e��5m=�w�o}��,����eJ�]w쳲t�o{x�ƛ{X7�!=f�ϱIc۝���������0|�s�&�p��k�� H�tVMr55ܿS�b,[����8B Western Blot washing buffer: 1X PBST (Phosphate-buffered saline, 0.1% Tween 20) For 1.0 L: 100 mL 10X PBS 899 mL ddH 2 O 1 mL Tween 20: 1X TBST (Tris-buffered saline, 0.1% Tween 20) For 1.0 L: 100 mL 10X TBS 899 mL ddH 2 O 1 mL Tween 20: Blocking buffer: Western Blot blocking buffer: 5% (or 3%) nonfat dried milk or BSA in 1X TBST or 1X PBST. trailer <<1F1593BFCF224E79865E3332E1712407>]/Prev 366405>> startxref 0 %%EOF 148 0 obj <>stream 0000015072 00000 n 0000000956 00000 n ��_Uܞnٞ��Ae�ҒZRK��"��~�4F?ހji�[N��%�����4�d֏��&� [5e���2��Fŏ'��3���Vs*�j. Ensure the volume of the antibody solution is enough to fully cover the membrane. 116 0 obj <> endobj xref 0000004985 00000 n }2�N�Fͷ��M��k�_g�R�y;}�����hb6ʪ�/�����j2�:c��Qq'�0�*ː��{5Y ~^�&/��N[�7j�ֶT�{�B��p��2��V�ܪa��Z U�v)��e-w��67���odLli�c48�Y�i��{�~��?ڢ�|��YY�+�f�I�4�~`TfsK���l �v��] "|������5ɇMn�ԃr)�Βq�ޒrk�r��@��zI> �Ag����n�:�-W���� C�hz�;|�'y4�t.�ɯ��x3m��F�����d�����������7�j ����=��A�M�٪�j���8�Op6� �c��&nO9�{��~�6�ù����>��֌]�pu��}�x�(^� �d�����^���]YU#��xDk�C����d *C��0 T�d �7Jb�܇>����2��Xٙ5�>D][� H��VMo$5����q�0^��-�"��V�2H,�edQ�!+���Wnw�lr �4�g�����>~=�u��24s��i�N$폿O���x�����/NO���o�~z}uyu�k��7_�ig���-�����Qˏ;�{���{���~0o�L��}�?N}��ks�? 0000004280 00000 n 0000030420 00000 n �j��L}�A��ݱ�0uV,/��O�u�fVe���z&#��b�@x큽{�O�l7�����K��!��KSTZ�~Z�u?7�x�ґ����L��X���%G�J]���I�F'��e�(�R"�`,�1"K��Q%iJP�1n�[ƯI��o8:�[[email protected][�F�$V���_���"}�T2�J�ڦ�4#!Pz�m�m�/��BւBϏڊFO�\x��s�E�[>�8D>��iޤ[email protected] ��(�lt7f�g.�]l~�G ����KەT�]�)z����ɓ���]�|���B�Ộ�_K����W �����^�����g ,��JEm�Q�I��_.����~#F]�oZ�Y_�{T_.�a=�S$����X2�h8c�N[׫�=��G�g�:�ѹ�'���I���b���M�J�t/�RZl��r�Τ��n��ފױ��m*�݅�6���:��I/ϻ��)�C�jk�}�nZ�ۮI`�N�-�����͓4�v����^?W]��K�?��M�/��ľۉ_�Һ��P�)� ��>stream