Anti-c-Jun (phospho S63) antibody [Y172] - BSA and Azide free

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-c-Jun (phospho S63) antibody [Y172] - BSA and Azide free (AB227533)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A431 (Human epidermoid carcinoma cell line) cells labeling c-Jun (phospho S63) with ab32385 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing the expression was increased after treatment with anisomycin (1 µg/ml for 15 minutes), then decreased after treatment with the Lambda Protein Phosphatase treatment 31°C for 2 hours. The nuclear counter stain is DAPI (blue).
Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at a 1/200 dilution (red).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32385).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Jun (phospho S63) antibody [Y172] - BSA and Azide free (AB227533)

This IHC data was generated using the same anti-c Jun antibody clone, Y172, in a different buffer formulation (cat# ab32385).

ab32385, at a 1/50 dilution, staining c-Jun in paraffin embedded human breast carcinoma tissue by Immunohistochemistry.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Jun (phospho S63) antibody [Y172] - BSA and Azide free (AB227533)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast tissue sections labeling c-Jun with Purified ab32385 at 1 : 250 dilution (0.46 µg/ml). Heat mediated antigen retrieval was performed using using ab93684 (Tris/EDTA buffer, pH 9.0)ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody.Negative control : PBS instead of the primary antibody.Hematoxylinwas used as a counterstainThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32385).

• WB

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Western blot - Anti-c-Jun (phospho S63) antibody [Y172] - BSA and Azide free (AB227533)

Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-c-Jun (phospho S63) antibody [Y172] (<a href='/en-us/products/primary-antibodies/c-jun-phospho-s63-antibody-y172-ab32385'>ab32385</a>) at 0.1 µg/mL

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 15 µg

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) treated with 1 ug/ml anisomycin for 15 minutes whole cell lysates at 15 µg

Lane 3:

HeLa (Human cervix adenocarcinoma epithelial cell) treated with 1 ug/ml anisomycin for 15 minutes whole cell lysates 15ug. Then the membrane was incubated with phosphatase. at 15 µg

Secondary

All lanes:

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

Predicted band size: 35 kDa

false

• WB

Lab

Western blot - Anti-c-Jun (phospho S63) antibody [Y172] - BSA and Azide free (AB227533)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32385).

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

All lanes:

Western blot - Anti-c-Jun (phospho S63) antibody [Y172] (<a href='/en-us/products/primary-antibodies/c-jun-phospho-s63-antibody-y172-ab32385'>ab32385</a>) at 1/1000 dilution

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 15 µg

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) treated with 1ug/mL anisomycin for 15 minutes whole cell lysates at 15 µg

Lane 3:

HeLa (Human cervix adenocarcinoma epithelial cell) treated with 1ug/ml anisomycin for 15 minutes whole cell lysates. Then the membrane was incubated with phosphatase. at 15 µg

Secondary

All lanes:

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

Predicted band size: 35 kDa

false

• Dot

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Dot Blot - Anti-c-Jun (phospho S63) antibody [Y172] - BSA and Azide free (AB227533)

Unpurified ab32385 used at a 1 : 1000 dilution.
Secondary antibody is Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) used at a 1 : 100,000 dilution. Blocking/Diluting buffer and concentration : 5% NFDM/TBST.
Lane 1 : Human c-Jun (pS63) phospho peptide.
Lane 2 : Human c-Jun non-phospho peptide.
Exposure time 3 minutes.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32385).

• WB

Unknown

Western blot - Anti-c-Jun (phospho S63) antibody [Y172] - BSA and Azide free (AB227533)

Blocking and diluting buffer : 5% NFDM/TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32385).

All lanes:

Western blot - Anti-c-Jun (phospho S63) antibody [Y172] (<a href='/en-us/products/primary-antibodies/c-jun-phospho-s63-antibody-y172-ab32385'>ab32385</a>) at 0.1 µg/mL

Lane 1:

NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates at 15 µg

Lane 2:

NIH/3T3 (Mouse embryonic fibroblast) treated with 250 ng/ml anisomycin for 30 minutes whole cell lysates at 15 µg

Lane 3:

NIH/3T3 (Mouse embryonic fibroblast) treated with 250 ng/ml anisomycin for 30 minutes whole cell lysates. Then the membrane was incubated with phosphatase at 15 µg

Secondary

All lanes:

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

Predicted band size: 35 kDa

false

• ELISA

Unknown

ELISA - Anti-c-Jun (phospho S63) antibody [Y172] - BSA and Azide free (AB227533)

Antigen pS63 : c-Jun (phospho S63); NP : c-Jun non-phospho. Antigen concentration 0.01~1 ng/ml.

Primary antibody concentration range 0~1000 ng/ml.

Secondary antibody is an Alkaline Phosphatase-conjugated Goat Anti-Rabbit IgG(H+L) used at a 1 : 2500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32385).