Anti-ERK1 + ERK2 抗体

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 + ERK2 antibody (AB17942)

Immunohistochemistry analysis of ERK1/2 (pan) showing staining in the cytoplasm and nucleus of paraffin-embedded human breast carcinoma tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with ab17942 diluted in 3% BSA-PBS at a dilution of 1 : 100 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 + ERK2 antibody (AB17942)

Immunohistochemistry analysis of ERK1/2 (pan) showing staining in the cytoplasm and nucleus of paraffin-embedded human colon carcinoma tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with ab17942 diluted in 3% BSA-PBS at a dilution of 1 : 20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

• ICC

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Immunocytochemistry - Anti-ERK1 + ERK2 antibody (AB17942)

Immunofluorescent analysis of ERK1 + ERK2 Antibody was done on 70% confluent log phase U87-MG cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with ab17942 at 1 : 250 dilution in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor 488 Goat Anti-Rabbit IgG Secondary Antibody at a dilution of 1 : 400 for 45 minutes at room temperature (Panel a : green). Nuclei (Panel b : blue) were stained with SlowFade® Gold Antifade Mountant with DAPI. F-actin (Panel c : red) was stained with Alexa Fluor 594 Phalloidin. Panel d is a merged image showing cytoplasmic and nuclear localization. Panel e is a no primary antibody control. The images were captured at 40X magnification.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 + ERK2 antibody (AB17942)

Immunohistochemistry analysis of ERK1/2 (pan) showing staining in the cytoplasm and nucleus of paraffin-embedded mouse stomach tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with ab17942 diluted in 3% BSA-PBS at a dilution of 1 : 20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

• WB

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Western blot - Anti-ERK1 + ERK2 antibody (AB17942)

All lanes:

Western blot - Anti-ERK1 + ERK2 antibody (ab17942) at 1/1000 dilution

Lane 1:

K562 cells at 20 µg

Lane 2:

Daudi cells at 20 µg

Lane 3:

Hep G2 cells at 20 µg

Lane 4:

PC-3 cells at 20 µg

Lane 5:

NIH 3T3 cells at 20 µg

Secondary

All lanes:

Anti-Rabbit IgG - HRP Secondary Antibody at 1/5000 dilution

Predicted band size: 41 kDa

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• WB

AbReview11057****

Western blot - Anti-ERK1 + ERK2 antibody (AB17942)

The 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.

All lanes:

Western blot - Anti-ERK1 + ERK2 antibody (ab17942) at 1/1000 dilution

Lane 1:

Rat spinal cord tissue homogenate from animals that underwent Sham surgery at 20 µg

Lanes 2 - 3:

Rat spinal cord tissue homogenate from animals that underwent L5 nerve transection at 20 µg

Secondary

All lanes:

HRP conjugated goat anti-rabbit antibody at 1/3000 dilution

Predicted band size: 41 kDa

Observed band size: 42 kDa,44 kDa

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Exposure time: 5min

This image is courtesy of an anonymous Abreview

• WB

PubMed

Western blot - Anti-ERK1 + ERK2 antibody (AB17942)

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 4°C. 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

All lanes:

Western blot - Anti-ERK1 + ERK2 antibody (ab17942)

Predicted band size: 41 kDa

true

Image from PLoS One. 2014; 9(6): e99219. Fig3A, doi: 10.1371/journal.pone.0099219 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

• WB

Unknown

Western blot - Anti-ERK1 + ERK2 antibody (AB17942)

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.

All lanes:

Western blot - Anti-ERK1 + ERK2 antibody (ab17942)

Predicted band size: 41 kDa

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