82380

antibody from Invitrogen Antibodies

Targeting: MAPK3 ERK1, p44erk1, p44mapk, PRKM3

Western blot

Immunoprecipitation

Antibody data

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Validation data

MAPK3 protein structure - 82380 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of ERK1/ERK2 was performed using 70% confluent log phase RSC96 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with ERK1/ERK2 Rabbit Polyclonal Antibody (Product # 82380) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight 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 represents the merged image showing cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of ERK1/ERK2 was performed using 70% confluent log phase RSC96 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with ERK1/ERK2 Rabbit Polyclonal Antibody (Product # 82380) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (Heavy Chain) 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 represents the merged image showing cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 5 The effect of FGF2 on the phosphorylation of Akt and ERK1/2. (A) The phosphorylation of Akt and ERK1/2 was determined by immunoblot analysis. GAPDH was used as a loading control. Images are representative of five animals in each group. (B,C) The histograms show the normalized optical density analysis. The phosphorylation of Akt and ERK1/2 in the I/R group was transiently increased at 6 h after reperfusion compared to the Sham group, whereas there was no significant difference between the I/R group and the Sham group at 24 and 72 h. FGF2 treatment strikingly increased the phosphorylation of Akt and ERK1/2 at 24 and 72 h after reperfusion compared to the I/R group. Representative data of five individual samples in each group. * P < 0.05, ** P < 0.01, *** P < 0.001; ns indicates not statistically significant.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 6 The effect of FGF2 on the inhibition of apoptosis and ER stress and the activation of the Akt and ERK1/2 signaling pathway was detected in vitro . (A) The expression of pro-apoptotic proteins, the expression of ER stress-relevant proteins, and the phosphorylation of Akt and ERK1/2 were detected by Western blotting. Images are representative of five animals in each group. (B-I) The histograms show the normalized optical density analysis. FGF2 significantly decreased the expression of cleaved Caspase-3, Bax, CHOP, GRP78, ATF-6, and XBP-1 in NRK-52E treated with TBHP. The phosphorylation of Akt and ERK1/2 was increased in the TBHP-FGF2 group compared with the TBHP group. Representative data of five individual samples in each group. * P < 0.05, ** P < 0.01, *** P < 0.001; ns indicates not statistically significant.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 7 LY294002 and U0126 abolish the effect of FGF2 on the phosphorylation of Akt and ERK1/2 in vitro . (A) The phosphorylation of Akt and ERK1/2 was determined by Western blotting analysis. GAPDH was used as a loading control. Images are representative of five animals in each group. (B,C) The histograms show the normalized optical density analysis. LY294002 inhibited the effect of FGF2 treatment on the phosphorylation of Akt, whereas U0126 strikingly blocked the effect of FGF2 on the phosphorylation of ERK1/2. Representative data of five individual samples in each group. * P < 0.05, ** P < 0.01, *** P < 0.001; ns indicates not statistically significant.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5 The regulation effect of FGF10 on ER stress and ERK1/2 signaling pathway. (A) The expression levels of GRP-78, CHOP, XBP-1, ATF-4, ATF-6, PDI, ERK1/2, and phospho-ERK1/2 in kidney tissues of Sham group, I/R group and I/R-FGF10 group were determined by immunoblot analysis. FGF10 significantly increased the phosphorylation of ERK1/2. (B-I) The histograms show the normalized optical density analysis. Results are representative of five rats in each group. * P < 0.05, ** P < 0.01, *** P < 0.001, ns represents no significant difference.