- AHZ0422 - Invitrogen Antibodies CDKN1A antibody

Submitted references Proximal Tubule p53 in Cold Storage/Transplantation-Associated Kidney Injury and Renal Graft Dysfunction.

Xiang X, Zhu J, Zhang G, Ma Z, Livingston MJ, Dong Z
Frontiers in medicine 2021;8:746346

Hypoxia-inducible factor-2α mediates senescence-associated intrinsic mechanisms of age-related bone loss.

Lee SY, Park KH, Lee G, Kim SJ, Song WH, Kwon SH, Koh JT, Huh YH, Ryu JH
Experimental & molecular medicine 2021 Apr;53(4):591-604

Cardiomyocyte Contractility and Autophagy in a Premature Senescence Model of Cardiac Aging.

Häseli S, Deubel S, Jung T, Grune T, Ott C
Oxidative medicine and cellular longevity 2020;2020:8141307

A pro longevity role for cellular senescence.

Attaallah A, Lenzi M, Marchionni S, Bincoletto G, Cocchi V, Croco E, Hrelia P, Hrelia S, Sell C, Lorenzini A
GeroScience 2020 Jun;42(3):867-879

UHPLC-Q/Orbitrap/MS/MS fingerprinting and antitumoral effects of Prosopis strombulifera (LAM.) BENTH. queous extract on allograft colorectal and melanoma cancer models.

Persia FA, Troncoso ME, Rinaldini E, Simirgiotis M, Tapia A, Bórquez J, Mackern-Oberti JP, Hapon MB, Gamarra-Luques C
Heliyon 2020 Feb;6(2):e03353

Aurora-A overexpression is linked to development of aggressive teratomas derived from human iPS cells.

Ohmine S, Salisbury JL, Ingle J, Pettinato G, Haddox CL, Haddad T, Galanis E, Ikeda Y, D'assoro AB
Oncology reports 2018 Apr;39(4):1725-1730

G1 checkpoint is compromised in mouse ESCs due to functional uncoupling of p53-p21Waf1 signaling.

Suvorova II, Grigorash BB, Chuykin IA, Pospelova TV, Pospelov VA
Cell cycle (Georgetown, Tex.) 2016;15(1):52-63

Rapamycin induces pluripotent genes associated with avoidance of replicative senescence.

Pospelova TV, Bykova TV, Zubova SG, Katolikova NV, Yartzeva NM, Pospelov VA
Cell cycle (Georgetown, Tex.) 2013 Dec 15;12(24):3841-51

p21Waf1 is required for complete oncogenic transformation of mouse embryo fibroblasts by E1Aad5 and c-Ha-ras oncogenes.

Romanov VS, Bardin AA, Zubova SG, Bykova TV, Pospelov VA, Pospelova TV
Biochimie 2011 Sep;93(9):1408-14

Histone deacetylase inhibitor vorinostat suppresses the growth of uterine sarcomas in vitro and in vivo.

Hrzenjak A, Moinfar F, Kremser ML, Strohmeier B, Petru E, Zatloukal K, Denk H
Molecular cancer 2010 Mar 4;9:49

Pluripotent cell division cycles are driven by ectopic Cdk2, cyclin A/E and E2F activities.

Stead E, White J, Faast R, Conn S, Goldstone S, Rathjen J, Dhingra U, Rathjen P, Walker D, Dalton S
Oncogene 2002 Nov 28;21(54):8320-33

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of p21/Cip1 in PC12 cells using a p21/Cip1 monoclonal antibody (Product # AHZ0422).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Flow cytometry analysis of p21/ Cip1 was done on MCF7 cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with p21/ Cip1 Mouse Monoclonal Antibody (AHZ0422, red histogram) or with mouse isotype control (pink histogram) at 3-5 ug/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Rabbit Anti-Mouse Secondary Antibody (A11059) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2 Cellular senescence in cultured, neonatal cardiomyocytes. Assessed were discriminative biomarkers of cellular senescence in murine cardiac myocytes at indicated time points post primary cell isolation. Relative mRNA expressions of (a) Ki-67, (b) PCNA, and (c) p16 were quantified via qPCR analyses ( n = 4 mice). (d) Protein levels of p53 were determined by immunoblot analyses normalized to GAPDH, and representative blots are illustrated ( n = 4 mice). (e) The signal of immunofluorescently stained p21 in cardiomyocyte nuclei and (f) autofluorescence per cardiac myocytes were microscopically quantified ( n = 4 mice). (g) As validated biomarker of cellular senescence, the SA- beta -Gal activity at pH 6 was measured qualitatively as positively stained cardiomyocytes per total number of heart muscle cells ( n = 6 mice). Data are presented as mean values +- SD. Statistical significance was assessed by one-way ANOVA ( p < 0.05); a reference day 6; b reference day 9; c reference day 13; d reference day 17.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 5 Upregulation of HIF-2alpha expression by aging stimulates osteoclastogenesis and osteoclast senescence. a The mRNA levels of osteoclast marker genes ( Trap ), Hif-2 alpha, and senescence markers ( p16 , p21 ) during M-CSF/RANKL-induced osteoclastogenesis of bone marrow macrophages (BMMs) ( n >= 3). b Representative images of p16 and p21 immunostaining in bone osteoclasts from young (4-month-old) and old (12-month-old) mice. Scale bar: 25 mum. M month. c, d TRAP staining ( c ) and qRT-PCR analysis of Hif-2alpha , Trap , p16 , and p21 expression ( n >= 4; d ) in Hif-2 alpha-overexpressing osteoclasts. BMMs were infected with Ad-C or Ad- Hif-2alpha and cultured with M-CSF and RANKL for 5 days. Scale bar: 100 mum. e The mRNA levels of Hif-2alpha , Trap, p16 , and p21 . Osteoclasts isolated from Hif-2alpha fl/fl mice were infected with Ad-C or Ad- Cre during osteoclastogenesis ( n >= 4). f, g Binding of HIF-2alpha to the promoter regions of p16 ( f ) and p21 ( g ). Ad- Hif-2alpha -infected osteoclasts were subjected to ChIP with an anti-HIF-2alpha antibody and a primer pair designed to span the putative HIF-2alpha binding regions [5''-(A/G)CGTG-3''] within the promoters of p16 ( n = 3; f ) and p21 ( n = 3; g ). The values are presented as the means +- SDs (* P < 0.05, ** P < 0.01, and *** P < 0.005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 6 Osteoclast-specific depletion of HIF-2alpha increases bone mass in aged mice. a - c Immunostaining of HIF-2alpha, p16, and p21 in Hif-2alpha fl/fl and Hif-2alpha ; fl/fl Ctsk - Cre mice. Osteoclast-specific depletion of HIF-2alpha in 12-month-old Hif-2alpha fl/fl and Hif-2alpha ; fl/fl Ctsk - Cre mice was assessed by immunohistochemistry with anti-HIF-2alpha antibody. The dotted lines indicate osteoblasts (Scale bar: 25 mum; a ). Immunostaining of p16 ( b ) and p21 ( c ) in osteoclasts from Hif-2alpha fl/fl and Hif-2alpha ; fl/fl Ctsk - Cre mice was examined by immunofluorescence microscopy. Scale bar: 25 mum. d Quantitative uCT analysis of trabecular bones. BMD, BV/TV, Tb.Th, Tb.Sp, and Tb.N in trabecular bones from 12-month-old Hif-2alpha fl/fl and Hif-2alpha ; fl/fl Ctsk-Cre mice ( n = 8). e Representative images of H&E and TRAP staining in 12-month-old Hif-2alpha fl/fl and Hif-2alpha ; fl/fl Ctsk-Cre mice ( n = 8; Scale bar, 100 mum). BV/TV, N.Ob/B.Pm, Ob.S/BS, N.Oc/B.Pm, and Oc.S/BS were determined by bone histomorphometric analyses of the metaphyseal regions of femurs. The values are presented as the means +- SDs (* P < 0.05, ** P < 0.01, *** P < 0.005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5 Pifithrin-alpha protects kidneys during cold storage/transplantation. 2.2 mg/kg pifithrin-alpha or the vehicle solution DMSO was injected intraperitoneally to donor mice. After overnight treatment, donor kidneys were collected for 8.5 h of cold storage in UW solution in the presence of 50 muM pifithrin-alpha or DMSO. After kidney transplantation surgery, 2.2 mg/kg pifithrin-alpha or DMSO was added to the abdomen cavity of recipient mice. The transplanted kidneys were collected at 24 h after transplantation. The contralateral kidneys of donor mice were used as sham control. (A) Representative images of H&E staining of renal histology (Scale bar, 0.2 mm) and TUNEL assay (Scale bar, 0.1 mm). (B) Tubular damage score. (C) Quantification of TUNEL positive cells in outer medulla and cortical tissues. (D) Representative image of p53 and p-p53 (s15) immunofluorescence. Scale bars, 0.1 mm. Arrow: positive staining. (E,F) Quantification of p53 and p-p53 (s15) positive cells in outer medulla and cortical tissues. Quantitative data are expressed as mean +- SD (n >= 4). * P < 0.05 vs. sham control, # P < 0.05 vs. DMSO Cold 8.5 h/24 h. (G) Immunoblots analysis and quantification of p53, p-p53 (s15), p21 and PUMA with GAPDH as loading control.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 6 Ablation of p53 from proximal tubules in donor kidneys reduces cold-storage transplantation injury. Donor kidneys were collected from p53 proximal tubules knockout mice (KO) and their wild-type littermates (WT), stored in cold UW solution for 8.5 h, and transplanted into WT recipient mice. The transplanted kidneys were collected 24 h after transplantation for analysis. The contralateral kidney of donor was used as sham control. (A) Representative images of H&E staining. Scale bar, 0.2 mm. (B) Pathologic score of tubular damage. (C) Representative images of TUNEL assay. Scale bar, 0.1 mm. (D) Quantification of TUNEL positive cells in outer medulla and cortical tissues. (E) Representative image of p53 immunofluorescence staining. Scale bars, 0.1 mm. Arrow: positive staining. (F) Quantification of p53 positive cells in outer medulla and cortical tissues. (G) Representative image of p-p53 (s15) immunofluorescence staining. Scale bars, 0.1 mm. Arrow: positive staining. (H) Quantification of p-p53 (s15) positive cells in outer medulla and cortical tissues. Quantitative data are expressed as mean +- SD (n >= 4). * P < 0.05 vs. sham control, # P < 0.05 vs. WT Cold 8.5 h/24 h. (I) Immunoblots analysis and quantification of p53, p-p53 (s15), PUMA and p21 with GAPDH as loading control.

AHZ0422

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