Wayne L G, Diaz G A
Anal Biochem. 1986 Aug 15;157(1):89-92. doi: 10.1016/0003-2697(86)90200-9.
Mycobacteria produce two classes of catalase, designated T and M. Only the T-catalase also has a peroxidase-like function. When a 3,3'-diaminobenzidine (DAB) peroxidase stain was applied to polyacrylamide gel electrophoresis gels, followed by a ferricyanide negative stain for catalase, isoenzymes of T-catalase appeared as dark bands within a zone of clearing in the green background; the M-catalase appeared only as a clear zone. Heated and unheated preparations could be used to demonstrate the presence of comigrating bands of M and T. The application of the ferricyanide stain after the DAB stain of T-catalase resulted in marked intensification of the positive bands of T-catalase due to nonenzymatic, peroxide-independent reduction of the ferricyanide by the DAB product.
分枝杆菌产生两类过氧化氢酶，分别称为T型和M型。只有T型过氧化氢酶还具有类似过氧化物酶的功能。当将3,3'-二氨基联苯胺（DAB）过氧化物酶染色应用于聚丙烯酰胺凝胶电泳凝胶，随后用铁氰化物负染法检测过氧化氢酶时，T型过氧化氢酶的同工酶在绿色背景的透明区内呈现为深色条带；M型过氧化氢酶仅呈现为一个透明区。加热和未加热的制剂可用于证明M型和T型共迁移条带的存在。在对T型过氧化氢酶进行DAB染色后应用铁氰化物染色，由于DAB产物对铁氰化物的非酶促、不依赖过氧化物的还原作用，导致T型过氧化氢酶的阳性条带明显加深。
