- 44-1160G - Invitrogen Antibodies PPP1R2 antibody

Bresch AM, Yerich N, Wang R, Sperry AO
BMC molecular and cell biology 2020 Nov 25;21(1):84

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: HeLa cells grown on a chamber slide were fixed in cold 10% trichloroacetic acid for 10 minutes, rinsed in TBS and permeabilized with 0.1% Triton-X-100 for 15 minutes. Slides were incubated with the I-2 (pT72) antibody

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: HeLa cells grown on a chamber slide were fixed in cold 10% trichloroacetic acid for 10 minutes, rinsed in TBS and permeabilized with 0.1% Triton-X-100 for 15 minutes. Slides were incubated with the I-2 (pT72) antibody

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Phospho-Inhibitor-2 pThr72 was done on 70% confluent log phase A431 cells treated with 3uM of Nocodazole for 24 hours. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Phospho-Inhibitor-2 pThr72 Rabbit Polyclonal Antibody (Product # 44-1160G) at 1:250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing punctuated nuclear localization. Panel e is untreated cell with no signal. Panel f is a no primary antibody control. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of Phospho-Inhibitor-2 [pThr72] was done on HeLa 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 Phospho-Inhibitor-2 [pThr72] Rabbit Polyclonal Antibody (441160G, red histogram) or with rabbit isotype control (yellow 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 Goat Anti-Rabbit Secondary Antibody (A11008) 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)
Main image
Experimental details: Fig. 4 PP1, PPP1R2, pR2, and AURKA are localized at the midbody. a ARPE-19 cells were fixed and stained for the indicated endogenous proteins (green) localized relative to alpha-tubulin of the central spindle (red). b Average intensity from the green channel along the central spindle was measured in 10-0.87 mum 2 square regions spanning the length of the central spindle in 30 dividing cells and is plotted in ( c ). Size bar equals 5 mum. Asterisks indicate statistical significance compared to control (* p

44-1160G