Anti-PP2A alpha + beta 抗体 [EPR11787(B)]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-PP2A alpha + beta antibody [EPR11787(B)] (AB168350)

Immunofluorescent analysis of A431 cells labeling PP2A alpha + beta with ab168350 at 1/50 dilution.

• WB

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Western blot - Anti-PP2A alpha + beta antibody [EPR11787(B)] (AB168350)

All lanes:

Western blot - Anti-PP2A alpha + beta antibody [EPR11787(B)] (ab168350) at 1/10000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

293T (Human embryonic kidney epithelial cell) cell lysate at 10 µg

Lane 3:

A431 cell lysate at 10 µg

Lane 4:

NIH/3T3 cell lysate at 10 µg

Secondary

All lanes:

HRP labeled goat anti-rabbit at 1/2000 dilution

Predicted band size: 36 kDa

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• WB

Supplier Data

Western blot - Anti-PP2A alpha + beta antibody [EPR11787(B)] (AB168350)

Blocking and Diluting buffer and concentration : 5% NFDM/TBST.

Human PP2A beta full length recombinant protein contains aa1-309 with a GST-Tag (catalog# : ab128562).

Human PP2A alpha full length recombinant protein contains aa1-309 with a GST-Tag (catalog# : ab128557).

All lanes:

Western blot - Anti-PP2A alpha + beta antibody [EPR11787(B)] (ab168350) at 1/20000 dilution

Lane 1:

Human PP2A beta full length recombinant protein at 0.015 µg

Lane 2:

Human PP2A alpha full length recombinant protein at 0.015 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 36 kDa

Observed band size: 59 kDa

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Exposure time: 15s

• WB

Unknown

Western blot - Anti-PP2A alpha + beta antibody [EPR11787(B)] (AB168350)

All lanes:

Western blot - Anti-PP2A alpha + beta antibody [EPR11787(B)] (ab168350) at 1/10000 dilution

All lanes:

immunoprecipitation pellet from HeLa cell lysate

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG preferentially detecting the non-reduced form of rabbit IgG

Predicted band size: 36 kDa

false

• WB

Lab

Western blot - Anti-PP2A alpha + beta antibody [EPR11787(B)] (AB168350)

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PP2A alpha + beta antibody [EPR11787(B)] (ab168350) at 1/20000 dilution

Lane 1:

Western blot - Recombinant Human PPP2CB protein (<a href='/products/proteins-peptides/recombinant-human-ppp2cb-protein-ab128562'>ab128562</a>) at 0.015 µg

Lane 2:

Recombinant human PP2A-alpha protein (ab128557) at 0.015 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 36 kDa

Observed band size: 59 kDa

false

Exposure time: 15s

• WB

CiteAb

Western blot - Anti-PP2A alpha + beta antibody [EPR11787(B)] (AB168350)

Western Blotting using Anti-PP2A alpha + beta antibody [EPR11787(B)], ab168350. Publication image from Zhong, Y. et al., 2019, Nat Commun, 31586053. Legend direct from paper.

ATG binds to PP2A to enhance its activity. Drug Affinity Responsive Target Stability (DARTS) assay was performed to test the direct binding of ATG to PP2A in renal cells. a HEK293T cell lysates were pre-incubated with various concentrations of ATG as indicated at 25 °C for 1 h prior to digestion with pronase (0 or 1 : 1000 dilution) for 20 min. Lysates were then probed for PP2A expression, with GAPDH as a loading control. b Surface Plasmon Resonance (SPR) assay with Biacore showing the steady-state fit of binding between ATG and PP2A. c PP2A activity in HEK293T cells treated with various ATG concentrations as indicated for 2 h. Total PP2A activity is expressed as a relative fold change to no ATG treatment. n = 3 experiments. *P < 0.05 and ***P < 0.001 when compared with control by one-way ANOVA with Tukey’s post hoc analysis. d PP2A activity in conditionally immortalized human podocytes exposed to normal glucose and high mannitol (NG; 5 mM glucose and 25 mM mannitol) or high glucose (HG; 30 mM glucose) conditions for 24 h prior to treatment with ATG (10 µM) or vehicle for 2 h. Total PP2A activity is expressed as a relative fold change to NG vehicle control. ***P < 0.001 compared wqith vehicle control; n = 3. e PP2A activity in isolated glomeruli of control and diabetic eNOS−/− mice. Total PP2A activity is expressed as a relative fold change to vehicle-treated nondiabetic control. n = 3 per group. **P < 0.01 and ***P < 0.001 compared with vehicle control; #P < 0.05 compared wqith vehicle-treated control mice. f PP2A activity in isolated glomeruli of db/m and db/db mice. Total PP2A activity is expressed as a relative fold change to vehicle-treated db/m mice. n = 5 per group. *P < 0.05 and ***P < 0.001 compared with respective vehicle control; #P < 0.05 compared with vehicle-treated db/m mice by two-way ANOVA with Tukey’s post hoc analysis. *P < 0.05 and ***P < 0.001 when compared with control by two-way ANOVA with Tukey’s post hoc analysis. The data are represented as mean ± SD. g Computational prediction of ATG binding to PP2A crystal structure. Docking values for ATG for the binding using the virtual screening : Value S = −8.7414; E configuration = 3.2185; E place = -51.5684; E score1 = −8.7414. Source data are provided as a Source Data file

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• WB

CiteAb

Western blot - Anti-PP2A alpha + beta antibody [EPR11787(B)] (AB168350)

Western Blotting using Anti-PP2A alpha + beta antibody [EPR11787(B)], ab168350. Publication image from Zhong, Y. et al., 2019, Nat Commun, 31586053. Legend direct from paper.

ATG reduces TNF-α-induced inflammation through PP2A activation. a Levels of p65 NF-κB phosphorylation (p-p65) in the kidney glomeruli of control and diabetic mice with or without ATG treatment. Quantification of p-p65 normalized to total p65 (t-p65) is shown on the right. n = 3 per group. **P < 0.01 vs. nondiabetic mice, #P < 0.05 vs. vehicle-treated diabetic mice by two-way ANOVA with Tukey’s post hoc analysis. b Levels p-p65 in cultured podocytes treated with or without TNF-α. Quantification of p-p65 normalized to total p65 is shown on the right, n = 3. ***P < 0.001 vs. 0 µM ATG. c Overexpression of PP2A catalytic subunit (PP2Ac) by transient transfection reduces p65 NF-κB activation in cultured podocytes in vitro. EV, empty vector-transfected. Quantification of PP2A levels normalized to GAPDH and p-p65 normalized to t-p65 is shown on the right. ***P < 0.001 vs. EV; ###P < 0.001 vs. TNF-α-treated EV. d Levels p-p65 in cultured podocytes treated with ATG in presence or absence of okadaic acid (OA). Quantification of p-p65 levels normalized to GAPDH is shown on the right, n = 3. ***P < 0.001 vs. 0 µM ATG; ###P < 0.001 vs. TNF-α-treated EV by 1-way ANOVA with Tukey’s post hoc analysis. The data are represented as mean ± SD. Source data are provided as a Source Data file

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