Protocol to detect senescence-associated β-galactosidase and immunoperoxidase activity in fresh-frozen murine tissues

被引：1

作者：

Idelfonso-Garcia, Osiris German ^{[1
,2
]}

Pacheco-Rivera, Ruth ^{[3
]}

Alarcon-Sanchez, Brisa Rodope ^{[1
,4
]}

Serrano-Luna, Jesas ^{[4
]}

Baltierrez-Hoyos, Rafael ^{[5
,6
]}

Vasquez-Garzon, Veronica Rocio ^{[5
,6
]}

Muriel, Pablo ^{[7
]}

Villa-Trevino, Saal ^{[4
]}

Perez-Carreon, Julio Isael ^{[1
]}

Arellanes-Robledo, Jaime ^{[1
,6
]}

机构：

[1] Natl Inst Genom Med INMEGEN, Lab Liver Dis, Mexico City 14610, Mexico

[2] Metropolitan Autonomous Univ Iztapalapa Campus UAM, Dept Hlth Sci, Mexico City 09340, Mexico

[3] Natl Polytech Inst, Natl Sch Biol Sci, Dept Biochem, Lab Mol Diagnost, Mexico City 07738, Mexico

[4] Natl Polytech Inst CINVESTAV IPN, Ctr Res & Adv Studies, Dept Cell Biol, Mexico City 07360, Mexico

[5] Benito Juarez Autonomous Univ Oaxaca, Fac Med & Surg, Lab Fibrosis & Canc, UABJO, Mexico City 68120, Mexico

[6] Natl Council Humanities Sci & Technol CONAHCYT, Humanist & Sci Res, Mexico City 03940, Mexico

[7] Natl Polytech Inst CINVESTAV IPN, Ctr Res & Adv Studies, Dept Pharmacol, Lab Expt Hepatol, Mexico City 07360, Mexico

来源：

STAR PROTOCOLS | 2024年 / 5卷 / 02期

关键词：

Antibody; Health Sciences; Microscopy;

D O I：

10.1016/j.xpro.2024.103009

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Double labeling to identify different markers in the same tissue section represents a useful tool either for in situ diagnosis or characterization of molecular associations. Here, we present a protocol to detect senescence -associated 13-galac- tosidase (SA-13Gal) and immunoperoxidase (IPO) activity in fresh -frozen murine tissues. We describe steps for tissue collection, solution preparation, SA-13Gal staining, IPO staining, hematoxylin counterstaining, microscopic observation, and signal quantification. This protocol can be used to detect in situ proteins alongside SA-13Gal activity. For complete details on the use and execution of this protocol, please refer to Pacheco -Rivera et al.1

引用

页数：13

共 31 条

[21] Senescence-associated beta galactosidase (SABGAL)activity in normal, metaplastic and dysplastic mucosae of the upper GI tract.
Going, JJ
Stuart, RC
Monaghan, A
Downey, MG
Keith, WN
GASTROENTEROLOGY, 2000, 118 (04) : A275 - A275
[22] No increase in senescence-associated β-galactosidase activity in Werner syndrome fibroblasts after exposure to H2O2
de Magalhaes, JP
Migeot, V
Mainfroid, V
de Longueville, F
Remacle, J
Toussaint, O
STRATEGIES FOR ENGINEERED NEGLIGIBLE SENESCENCE: WHY GENUINE CONTROL OF AGING MAY BE FORESEEABLE, 2004, 1019 : 375 - 378
[23] Senescence-associated β-galactosidase activity and other markers of senescence are present in human peripheral blood CD8+cells during healthy aging
Dewald, Hannah K.
Martinez-Zamudio, Ricardo Ivan
Vasilopoulos, Themistoklis
Radeny, Joycelyn
Herbig, Utz
Fitzgerald-Bocarsly, Patricia
JOURNAL OF IMMUNOLOGY, 2021, 206
[24] USE OF DEXTRAN AND POST-PRIMARY ANTIBODY FIXATION IN IMMUNOPEROXIDASE STAINING OF FRESH-FROZEN TISSUE - DETECTION OF IMMUNOGLOBULIN ASSOCIATED WITH SQUAMOUS CARCINOMAS OF THE HEAD AND NECK
PANKOW, ML
DAVIS, LE
BECKER, SP
OSSOFF, RH
ANDERSON, BE
JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY, 1984, 32 (07) : 771 - 777
[25] TGF-β cytokines increase senescence-associated beta-galactosidase activity in human prostate basal cells by supporting differentiation processes, but not cellular senescence
Untergasser, G
Gander, R
Rumpold, H
Heinrich, E
Plas, E
Berger, P
EXPERIMENTAL GERONTOLOGY, 2003, 38 (10) : 1179 - 1188
[26] Reduction of senescence-associated beta-galactosidase activity by vitamin E in human fibroblasts depends on subjects' age and cell passage number
Ricciarelli, Roberta
Azzi, Angelo
Zingg, Jean-Marc
BIOFACTORS, 2020, 46 (04) : 665 - 674
[27] Senescence-associated β-galactosidase staining in fish cell lines and primary cultures from several tissues and species, including rainbow trout coelomic fluid and milt
Nguyen T. K. Vo
Michael S. Mikhaeil
Lucy E. J. Lee
Phuc H. Pham
Niels C. Bols
In Vitro Cellular & Developmental Biology - Animal, 2015, 51 : 361 - 371
[28] Senescence-associated β-galactosidase staining in fish cell lines and primary cultures from several tissues and species, including rainbow trout coelomic fluid and milt
Vo, Nguyen T. K.
Mikhaeil, Michael S.
Lee, Lucy E. J.
Pham, Phuc H.
Bols, Niels C.
IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL, 2015, 51 (04) : 361 - 371
[29] A rapid-response near-infrared fluorescent probe with a large Stokes shift for senescence-associated β-galactosidase activity detection and imaging of senescent cells
Qiu, Wenjing
Li, Xiaokang
Shi, Donglei
Li, Xinming
Gao, Ying
Li, Jinwen
Mao, Fei
Guo, Yuan
Li, Jian
Dyes and Pigments, 2020, 182
[30] A rapid-response near-infrared fluorescent probe with a large Stokes shift for senescence-associated β-galactosidase activity detection and imaging of senescent cells
Qiu, Wenjing
Li, Xiaokang
Shi, Donglei
Li, Xinming
Gao, Ying
Li, Jinwen
Mao, Fei
Guo, Yuan
Li, Jian
DYES AND PIGMENTS, 2020, 182

← 1 2 3 4 →