- PA5-20379 - Invitrogen Antibodies CDKN2A antibody

Submitted references IκB-ζ signaling promotes chondrocyte inflammatory phenotype, senescence, and erosive joint pathology.

Arra M, Swarnkar G, Alippe Y, Mbalaviele G, Abu-Amer Y
Bone research 2022 Feb 11;10(1):12

Senescence-associated-β-galactosidase staining following traumatic brain injury in the mouse cerebrum.

Tominaga T, Shimada R, Okada Y, Kawamata T, Kibayashi K
PloS one 2019;14(3):e0213673

Differential expression of cell cycle regulators in CDK5-dependent medullary thyroid carcinoma tumorigenesis.

Pozo K, Hillmann A, Augustyn A, Plattner F, Hai T, Singh T, Ramezani S, Sun X, Pfragner R, Minna JD, Cote GJ, Chen H, Bibb JA, Nwariaku FE
Oncotarget 2015 May 20;6(14):12080-93

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of mouse colon tissue lysate using a CDKN2A polyclonal antibody (Product # PA5-20379) at (A) 1 and (B) 2 µg/mL.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot Validation in Human Cell Lines. Loading: 15 µg of lysates per lane. Antibodies: p16INK4a Polyclonal Antibody (Product # PA5-20379) (2 µg/mL), 1h incubation at RT in 0.05 NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10,000 dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-p16INK4a Polyclonal Antibody (Product # PA5-20379) and a ~18 kDa band corresponding to CDKN2A was observed across cell lines tested . Whole cell extracts (30 µg lysate) of HeLa (Lane 1), HEK-293 (Lane 2), Hep G2 (Lane 3), PC-3 (Lane 4), HEL 92.1.7 (Lane 5) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0341BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1 µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:20000) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Atto Ultimate Sensitivity Substrate (Product # A38556). A non specific band (*) was observed around 40 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of p16INK4a in 293 WT or CDKN2A KO cells. Lysates (10 µg) were loaded onto SDS-PAGE and blots were probed with p16INK4a Polyclonal Antibody (Product # PA5-20379) diluted to 2 µg/mL and anti-beta actin diluted to 1 µg/mL. 1 h incubation at RT in 0.05 NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10,000 dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of recombinant proteins. Loading: 15 µg of lysate per lane. p16INK4a Polyclonal Antibody (Product # PA5-20379) (2 µg/mL), overnight incubation at 4˚C in 0.05 NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10,000 dilution. Lane 1: Human prostate (benign hyperplasia) Lane 2: Human CDKN2A recombinant protein (arrow: monomer and dimer)

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot Validation in Human Cell Lines. Loading: 15 µg of lysates per lane. Antibodies: p16INK4a Polyclonal Antibody (Product # PA5-20379) (2 µg/mL), 1h incubation at RT in 0.05 NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10,000 dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot Validation in Human Normal and Cancer Tissue. Loading: 15 µg of lysates per lane. Antibodies: p16INK4a Polyclonal Antibody (Product # PA5-20379) (2 µg/mL), overnight incubation at 4˚C in 0.05 NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10,000 dilution. Lane 1: Human breast Lane 2: Human lung tumor Lane 3: Human colon cancer

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot Validation in Mouse Colon Tissue. Loading: 15 µg of lysates per lane. Antibodies: p16INK4a Polyclonal Antibody (Product # PA5-20379) (A: 1 µg/mL, B: 2 µg/mL), 1h incubation at RT in 0.05 NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10,000 dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of human colon cells using a CDKN2A polyclonal antibody (Product # PA5-20379) at a 20 µg/mL dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of 4% paraformaldehyde-fixed A431 cells labeling CDKN2A with p16INK4a Polyclonal Antibody (Product # PA5-20379) at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1:500 dilution (green) and DAPI staining (blue).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of CDKN2A was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with CDKN2A/p16INK4a Polyclonal Antibody (Product # PA5-20379) at 1:100 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nucleus and cytoplasm localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 40X. magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of 4% paraformaldehyde-fixed human colon tissue labeling CDKN2A with p16INK4a Polyclonal Antibody (Product # PA5-20379) at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1:500 dilution (green) and DAPI staining (blue).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of 4% paraformaldehyde-fixed human colon tissue labeling CDKN2A with p16INK4a Polyclonal Antibody (Product # PA5-20379) at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1:500 dilution (green) and DAPI staining (blue).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of 4% paraformaldehyde-fixed Rat Colon Tissue labeling CDKN2A with p16INK4a Polyclonal Antibody (Product # PA5-20379) at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1:500 dilution (red) and DAPI staining (blue).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of paraffin-embedded Human Colon Tissue using p16INK4a Polyclonal Antibody (Product # PA5-20379) at 10 µg/mL. Tissue was fixed with formaldehyde and blocked with 0.1 serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of paraffin-embedded Rat Colon Tissue using p16INK4a Polyclonal Antibody (Product # PA5-20379) at 5 µg/mL. Tissue was fixed with formaldehyde and blocked with 0.1 serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 3 Evaluation of the effects of p25-GFP overexpression on cell cycle protein expression in malignant versus benign mouse MTC Immunoblots of lysates from proliferating malignant (P) and arrested benign (A) mouse MTC for A ) CDK2, CDK4 and CDK6; B ) cyclins A1, B1, D1, E2 and C ) p15 INK4b , p16 INK4a , p18 INK4c , p19 INK4d , p21 CIP/WAF1 and p27 KIP . P-values are p < 0.0001 for CDK2, p = 0.6974 for CDK4, p = 0.0633 for CDK6; p < 0.0001 for cyclin-A1, p = 0.0194 for cyclin-B1, p = 0.3416 for cyclin-D1, p < 0.0001 for cyclin-E2; p = 0.0002 for p15 INK4b , p = 0.291 for p16 INK4a , p = 0.0006 for p18 INK4c , p = 0.0054 for p19 INK4d , p = 0.02 for p21 CIP/WAF and p = 0.67 for p27 KIP1 . Data are represented as mean +/- SEM, N = 6 for each condition.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 4 Evaluation of the effect of CDK5 activity on cell cycle protein expression in human MTC cells Immunoblots of lysates from MTC-SK cells transfected with either a control plasmid (Con) or a kinase-dead (Kd) CDK5 encoding plasmid for A ) CDK2, CDK4 and CDK6; B ) cyclin-A1,-B1,-E2 and C ) p15 INK4b , p16 INK4a , p18 INK4c , p19 INK4d , p21 CIP/WAF1 and p27 KIP . P-values are p = 0.0244 for CDK2, p = 0.038 for CDK4, p = 0.009 for CDK6; p < 0.0001 for cyclin-A1, p = 0.0005 for cyclin-B1, p = 0.0116 for cyclin-E2; p = 0.0044 for p15 INK4b , p = 0.4687 for p16 INK4a , p = 0.1032 for p18 INK4c , p = 0.0195 for p19 INK4d , p = 0.0022 for p21 CIP/WAF and p = 0.2674 for p27 KIP1 . Data are represented as mean +/- SEM, N = 4-6 for each condition.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5 Analysis of cell cycle protein expression in human MTC samples Representative immunoblots of lysates from control thyroid tissue (Co), sporadic (Sp) and hereditary (H) human MTC tumors with antibodies as indicated are shown with quantification. Protein levels were normalized to Coomassie blue (CB) signal. Immunoblots were probed with antibodies to A ) CDK2, CDK4 and CDK6; B ) cyclins-A1, -D1, -E2; C ) in p15 INK4b , p16 INK4a , p18 INK4c , p19 INK4d , p21 CIP/WAF1 and p27 KIP . P-values were for CDK2, Sp, p = 0.0168 and H, p = 0.0140; for CDK4, Sp, p = 0.4072 and H, p = 0.8576; for CDK6, Sp, p = 0.5484 and H, p = 0.4916; for cyclin-A1, Sp, p = 0.9923 and H, p = 0.7017; for cyclin-D1, Sp, p = 0.0307 and H, p = 0.6883; for cyclin-E2, Sp, p = 0.2645 and H, p = 0.3070; for p15 INK4b , Sp, p = 0.0088 and H, p = 0.0310; for p16 INK4a , Sp, p = 0.0014 and H, p = 0.0496; for p18 INK4c , Sp, p = 0.0309 and H, p = 0.1054; for p19 INK4d , Sp, p = 0.0484 and H, p = 0.3560; for p21 CIP/WAF , Sp, p = 0.0554 and H, p = 0.6807; for p27 KIP1 , Sp, p = 0.2149 and H, p = 0.2131. Data are represented as mean +/- SEM, N = 4 for each condition.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig 4 Cyclin-dependent kinase inhibitor 2A (p16) immunohistochemistry and mRNA expression in the cerebrum 1 to 14 days after injury. (A) p16 immunostained cells could be observed in the ipsilateral hemicerebrum at 4, 7, and 14 days after injury in injury groups. Long scale = 50 mum, short scale = 12.5 mum. (B) Double immunohistochemistry of the ipsilateral hemicerebrum at 7 days after injury for co-localization of p16, glial fibrillary acidic protein (GFAP), DAPI (4',6-diamidino-2-phenylindole), and merge. Scale = 30 mum. (C) Graph of the numbers of p16 immunostained cells in the ipsilateral hemicerebrum in (A). (D-F) p16 mRNA expression in the ipsilateral hemicerebrum 1 to 14 days after injury. p16 mRNA expressions were normalized to that of glyceraldehyde-3-phosphate dehydrogenase ( Gapdh ) (D), hypoxanthine guanine phosphoribosyl transferase ( Hprt ) (E), and peptidylprolyl isomerase A ( Ppia ) (F) (* p < 0.05, n = 5 for control group, n = 5 for the sham group, n = 5 for the injury group).

PA5-20379

Antibody data