Anti-c-Jun (phospho T91 + T93 + T95) antibody [RP23040155] - ChIP Grade

• ChIP

Supplier Data

ChIP - Anti-c-Jun (phospho T91 + T93 + T95) antibody [RP23040155] - ChIP Grade (AB308130)

Enrichment of endogenous c-Jun (phospho T91 + T93 + T95) protein at specific gene loci using ab308130 : Chromatin Immunoprecipitation (ChIP) was performed using ab308130, 5 μg on sheared chromatin from 2 million U-937 cells and vs U-937 cells treated with 250 ng/mL anisomycin for 30 minutes using the "MAGnify ChIP system" kit. Normal Rabbit IgG (1 μg) was used as a negative IP control. The purified DNA was analyzed by 7500 Fast qPCR system with optimized PCR primer pairs for the promoters of the active JAG1, MMP13 region used as positive control target genes, and the region of the inactive SAT2 satellite repeat, used as negative control target gene. Data is compared between untreated vs treated samples and is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

• WB

Supplier Data

Western blot - Anti-c-Jun (phospho T91 + T93 + T95) antibody [RP23040155] - ChIP Grade (AB308130)

Western blot analysis was performed on whole cell extracts (30 µg lysate) of COLO 205 (Lane 1), U-937 (Lane 2), PC-3 (Lane 3) and U-87 MG (Lane 4). The blots were probed with ab308130, 1-2 µg/mL and detected by chemiluminescence using Goat antiRabbit IgG (H+L) Secondary Antibody, HRP conjugate (0.4 µg/mL, 1 : 2500 dilution). Two Isoforms of 43 and 36 kDa bands corresponding to c-Jun (phospho T91 + T93 + T95) was observed across cell lines tested. Known quantity of protein samples were electrophoresed using a 4-12% Bis-Tris gel, electrophoresis system and pre-stained protein standard. Resolved proteins were then transferred onto a nitrocellulose membrane. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed (ECL).

All lanes:

Western blot - Anti-c-Jun (phospho T91 + T93 + T95) antibody [RP23040155] - ChIP Grade (ab308130) at 1 µg/mL

Lane 1:

whole cell extracts of COLO 205 cells at 30 µg

Lane 2:

Whole cell extracts of U-937 cells at 30 µg

Lane 3:

Whole cell extracts of PC-3 cells at 30 µg

Lane 4:

whole cell extracts of U-87 MG cells at 30 µg

Secondary

All lanes:

Goat antiRabbit IgG (H+L) Secondary Antibody, HRP conjugate at 1/2500 dilution

Predicted band size: 36 kDa

Observed band size: 36 kDa

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