Anti-ARPC5/p16 ARC antibody [EP1551Y]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-ARPC5/p16 ARC antibody [EP1551Y] (AB51243)

Immunocytochemistry/Immunofluorescence analysis of Neuro-2a cells labelling p16 ARC with purified ab51243 at a dilution of 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

Control 1 : primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000).

Control 2 : ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ARPC5/p16 ARC antibody [EP1551Y] (AB51243)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen tissue labelling p16 ARC with purified ab51243 at a dilution of 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ARPC5/p16 ARC antibody [EP1551Y] (AB51243)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human uterus adenocarcinoma tissue labelling p16 ARC with unpurified ab51243 at a dilution of 1/50.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ARPC5/p16 ARC antibody [EP1551Y] (AB51243)

Immunohistochemical analysis of formalin fixed paraffin embedded human spleen labelling ARPC5/p16 ARC with ab51243 at a concentration of 0.1µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins ab51243 Anti-ARPC5/p16 ARC antibody [EP1551Y] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• IP

Unknown

Immunoprecipitation - Anti-ARPC5/p16 ARC antibody [EP1551Y] (AB51243)

ab51243 (purified) at a dilution of 1/20 immunoprecipitating p16 ARC in human fetal brain tissue lysate.

Lane 1 (input) : Human fetal brain tissue lysate (10μg)

Lane 2 (+) : ab51243 + human fetal brain tissue lysate.

Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab51243 in human fetal brain tissue lysate.

For western blotting, a HRP-conjugated anti-rabbit specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).

Blocking buffer and concentration : 5% NFDM/TBST.

Diluting buffer and concentration : 5% NFDM /TBST.

All lanes:

Immunoprecipitation - Anti-ARPC5/p16 ARC antibody [EP1551Y] (ab51243)

Predicted band size: 16 kDa

Observed band size: 16 kDa

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• WB

Lab

Western blot - Anti-ARPC5/p16 ARC antibody [EP1551Y] (AB51243)

Blocking and dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-ARPC5/p16 ARC antibody [EP1551Y] (ab51243) at 1/5000 dilution

Lane 1:

HeLa whole cell lysate at 20 µg

Lane 2:

SKBR-3 whole cell lysate at 20 µg

Lane 3:

MCF-7 whole cell lysate at 20 µg

Lane 4:

HepG2 whole cell lysate at 20 µg

Secondary

All lanes:

Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 16 kDa

Observed band size: 16 kDa

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• WB

Lab

Western blot - Anti-ARPC5/p16 ARC antibody [EP1551Y] (AB51243)

Blocking and dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-ARPC5/p16 ARC antibody [EP1551Y] (ab51243) at 1/5000 dilution

Lane 1:

Human fetal brain tissue lysate at 20 µg

Lane 2:

Mouse brain tissue lysate at 20 µg

Lane 3:

Rat brain tissue lysate at 20 µg

Secondary

All lanes:

Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 16 kDa

Observed band size: 16 kDa

false

• WB

Supplier Data

Western blot - Anti-ARPC5/p16 ARC antibody [EP1551Y] (AB51243)

Blocking buffer and concentration : 5% NFDM/TBST

Diluting buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-ARPC5/p16 ARC antibody [EP1551Y] (ab51243) at 1/2000 dilution

Lane 1:

Human fetal brain tissue lysate at 10 µg

Lane 2:

Mouse brain tissue lysate at 10 µg

Lane 3:

Rat brain tissue lysate at 10 µg

Secondary

All lanes:

Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 16 kDa

false

• WB

Lab

Western blot - Anti-ARPC5/p16 ARC antibody [EP1551Y] (AB51243)

Lane 1 : Wild type HAP1 whole cell lysate (20 μg)
Lane 2 : p16 ARC knockout HAP1 whole cell lysate (20 μg)
Lane 3 : HeLa whole cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab51243 observed at 16 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab51243 was shown to specifically react with p16 ARC when p16 ARC knockout samples were used. Wild-type and p16 ARC knockout samples were subjected to SDS-PAGE. ab51243 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-ARPC5/p16 ARC antibody [EP1551Y] (ab51243)

Predicted band size: 16 kDa

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• WB

CiteAb

Western blot - Anti-ARPC5/p16 ARC antibody [EP1551Y] (AB51243)

Western Blotting using Anti-ARPC5/p16 ARC antibody [EP1551Y], ab51243. Publication image from Li, Y. et al., 2021, Theranostics, 33391511. Legend direct from paper.

hUC-MSCs treatment normalizes markers of senescence in MRL/lpr mice splenic CD4+ T cells in vivo. (A, B) qPCR analysis showing the expression levels of p21, p16 in WT mice, PBS-treated MRL/lpr mice and hUC-MSCs-treated MRL/lpr mice. (C) Representative western blot showing the expression levels of p21, p16, p53 and acetylation of p53 in WT mice, PBS-treated MRL/lpr mice and hUC-MSCs-treated MRL/lpr mice. (D, E) Capillary WES and qPCR analysis showing the expression of Sirt1 in WT mice, PBS-treated MRL/lpr mice and hUC-MSCs-treated MRL/lpr mice. GAPDH was used as a protein loading control. *p < 0.05; **p < 0.01; ***p < 0.001.

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• WB

CiteAb

Western blot - Anti-ARPC5/p16 ARC antibody [EP1551Y] (AB51243)

Western Blotting using Anti-ARPC5/p16 ARC antibody [EP1551Y], ab51243. Publication image from Li, Y. et al., 2021, Theranostics, 33391511. Legend direct from paper.

Sirt1 is a mediator of hUC-MSCs increasing senescence of splenic CD4+ T cells in MRL/lpr mice. (A) Western blotting showing the expression of Sirt1, p21, p16, p53 and acetylation of p53 in EX527 and DMSO treated MRL/lpr splenic CD4+ T cells. (B) MRL/lpr splenic CD4+ T cells were pretreated with SRT1720 12 h before exposed to hUC-MSCs for 24-48 h, western blotting showing the expression of p21, p16, p53 and acetylation of p53. GAPDH was used as a protein loading control. All experimental data were verified in at least three independent experiments. *p < 0.05; **p < 0.01.

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