2. Antibodies

Autophagy Research
Anti SQSTM1/A170/p62, Rabbit
Wako Cat. No. 018-22141 (100 µL) <for Immunochemistry>
Keep at -20℃

Sequestosome 1 (SQSTM1) / A170 (mouse) / p62 (human) / ZIP (rat), a ubiquitin-binding protein, expresses oxidative stress-dependently.Abnormality of SQSTM1 leads to bone metabolic disorder, obesity and Type II diabetes. SQSTM1 was reported to bind LC3, which regulates autophagosome formation. The protein has attracted the attention of researchers because it is believed to induce a protein from ubiquitinproteosome system (UPS) to lysosome-dependent macroautophagy (autophagy) system, which are two major intracellular pathways for protein degradation.
Wako has launched the mouse SQSTM1 (A170) rabbit antiserum, which is applicable to Western blot, immunohistochemistry and immunofl uorescence.
Preparation:The antiserum is diluted with an equal amount of PBS and absorbed with E. coli proteins.
Immunogen:recombinant murine SQSTM1 (A170) (AA254-333) containing T7 tag at the N-terminal end and His tag at the carboxyterminal end
Specifi city:Specifi c for mouse and rat SQSTM1 (A170 / ZIP). Slightly reactive with human SQSTM1 (p62).
Working Dilution:Western blot 1 : 200 ; Immunohistochemistry 1 : 1,000 ; Immunofl uorescence 1 : 1,000
Figure:Western blot of cultivated mouse vascular smooth-muscle cell lysate (20µg)
Antibody:Anti SQSTM1/A170/p62 (Wako Cat. #018-22141) 1:200
Data was provided from Prof. Ishii, Tsukuba University (Japan)
Figure:Brain tissues were fixed with 4% paraformaldehyde and embedded with paraffin.The 6µm sections were stained with avidin-biotin-peroxidase method.
Primary Antibody:Anti SQSTM1/A170/p62 (Wako Cat. #018-22141) 1 : 1,000
Secondary Antibody:Anti rabbit IgG, biotin conjugated

1) Ishii, T., Yanagawa, T., Kawane, T., Yuki, K., Seita, J., Yoshida, H. and Bannai, S. : "Transcription factor Nrf2 coordinately regulates a group of oxidative stress-inducible genes in macrophages", Biochem. Biophys. Res. Commun., 226, 456-60 (1996).
2) Ishii, T., Itoh, K., Takahashi, S., Sato, H., Yanagawa, T., Katoh, Y., Bannai, S. and Yamamoto, M.: "Transcription factor Nrf2 coordinately regulates a group of oxidative stress-inducible genes in macrophages", J. Biol. Chem., 275, 16023-9 (2000).
3) Komatsu, M., Waguri, S., Koike, M., Sou, Y.S., Ueno, T., Hara, T., Mizushima, N., Iwata, J., Ezaki, J., Murata, S., Hamazaki, J., Nishito, Y., Ishii, T., Kobayashi, A., Yamamoto, M., Yue, Z., Uchiyama, Y., Kominami, E. and Tanaka, K.: "Homeostatic levels of p62 control cytoplasmic inclusion body formation in autophagy-defi cient mice", Cell, 131, 1149-63 (2007).
4) Nakaso, K., Kitayama, M., Ishii, T., Bannai, S., Yanagawa, T., Kimura, K., Nakashima, K., Ohama, E.. and Yamada, K. : "Eff ects of kainate-mediated excitotoxicity on the expression of rat counterparts of A170 and MSP23 stress proteins in the brain", Brain Res. Mol. Brain Res., 69, 155-63 (1999).
Research for NO
Anti soluble Guanylate Cyclase (sGC), MAb (Clone: mAB3221)
Wako Catalog No. 019-17801 (20 µg (40 µL)) <for Immunochemistry>
Keep at -20℃

Soluble guanylate cyclase (sGC), a hemoprotein, is the primary nitric oxide (NO) receptor in higher eukaryotes that catalyzes the conversion of guanosine 5'-triphosphate (GTP) to 3,5'-cyclic guanosine monophosphate (cGMP) and pyrophosphate (PPi) in the presence of Mg2+. The binding of NO to sGC leads to a several hundred-fold increase in cGMP synthesis.
Prepared from culture supernatant and prepared in glycine-Tris solution (pH 7.4). Contains no preservatives and stabilizers.
Isotype: IgG1
Specifi cally reacts with rat, bovine and human sGC, and strengthens in the reactivity on activation of sGC by NO, probably, due to the conformational changes of the enzyme and its associated antibody-antigen complex.
Working Dilution :
Westernblot 1 : 5,000 ; Immunofl uorescence 1 : 250
Tsuyama, S., et al., FEBS Lett., 455, 291 (1999).
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Wako Product Update Bio-No.1 [ page. 9 ]