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Our Abpromise guarantee covers the use of ab17942 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000. Predicted molecular weight: 42-44 kDa.|
This image is courtesy of an anonymous AbreviewThe tissue was harvested seven days post surgery, sonicated with RIPA buffer and the protein estimate made by Lowry. A 10% SDS-PAGE gel was run for 1.5 hr at 100V and transfered to PVDF membrane for 1.5 hr at 274 mA. The blot was blocked with 5% BSA for 1 hour at 23°C. The primary antibody was incubated with the blot for 18 hours at 4°C.
ab17942 staining ERK1 + ERK2 in Embryonic Stem Cell-induced Mouse Neurospheres by Immunohistochemistry (Frozen sections). Sections were formaldehyde-fixed prior to blocking in 10% serum for 30 minutes at room temperature. The primary antibody was diluted 1/1000 and incubated with the sample for 12 hours at 4°C. An Alexa Fluor® 594-conjugated Goat anti-Rabbit polyclonal antibody, diluted 1/1500 was used as the secondary.
ab17942 staining ERK1 + ERK2 in Human stomach tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 5% serum for 1 hour at 23°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/200 in blocking buffer) for 1 hourat 23°C. A HRP-conjugated GOat anti-rabbit IgG polyclonal was used as the secondary antibody.
ab17942 staining ERK1 + ERK2 in NIH3T3 Mouse Fibroblasts cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with PFA, permeabilized with 0.025% Triton-X in TBS and blocked with 5% serum for 1 hour at 23°C. Samples were incubated with primary antibody (1/200 in blocking buffer) for 16 hours at 4°C. An Alexa Fluor® 568-conjugated Goat anti-rabbit IgG polyclonal diluted to 1/1000 was used as the secondary antibody.
Blue channel - DAPI (nuclear staining)
Green channel - Alexa fluor 488 (B-Actin)
Image from PLoS One. 2014; 9(6): e99219. Fig3A, doi: 10.1371/journal.pone.0099219
Western blot analysis of Mice retinas (40-50μg/lane) labelling with anti-ERK1/2 at 1:300 (ab17942) and mouse monoclonal anti-phosphorylated ERK1/2 at 1:300 (ab50011), in 5% nonfat milk in TBST overnight at 4ºC. HRP conjugated antibodies were used as the secondary antibodies.
Data is expressed as percentage change in phosphorylated ERK1/2 (p-ERK1/2) over total ERK1/2 (t-ERK1/2) calculated in control and diabetic mice maintained with and without Edaravone treatment
Results are expressed as mean±SD. Values obtained from Normal group are considered as 100%. *P<0.05, ***P<0.001 vs. Normal, #P<0.05 vs. Edaravone
Western Blot for ab17942.
Extracts prepared from PC12 cells not stimulated (-), or stimulated with NGF (+) were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to nitrocellulose. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4C, then were incubated with ERK1&2 pan antibody for two hours at room temperature in a 3% BSA-TBST buffer. After washing, membranes were incubated
with goat anti-rabbit IgG alkaline phosphatase.
These data show that ab17942 ERK1&2 antibody allows the total amount of ERK1&2 to be measured.
Extracts prepared from PC12 cells not stimulated (-), or stimulated with NGF (+) were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to nitrocellulose. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4C, then were incubated with ERK1&2 pan antibody for two hours at room temperature in a 3% BSA-TBST buffer. After washing, membranes were incubated with goat anti-ra
ab17942 at 1/200 staining human (ab30203), rat (ab29480) and mouse (ab7261) kidney sections by IHC-P. The tissue sections were formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. An HRP conjugated goat anti-rabbit antibody was used as the secondary.
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