Anti-Histone H3 antibody [1B1B2] - Nuclear Marker and ChIP Grade

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 antibody [1B1B2] - Nuclear Marker and ChIP Grade (AB195277)

Immunofluorescent analysis of HeLa cells labeling Histone H3 with ab195277 at 1/500 dilution. Cells were fixed with 4% formaldehyde for 10 minutes and blocked with PBS/TX-100 containing 1% BSA. The left panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.

• ChIP

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ChIP - Anti-Histone H3 antibody [1B1B2] - Nuclear Marker and ChIP Grade (AB195277)

ChIP assays were performed using human HeLa cells, ab195277 and optimized PCR primer pairs for qPCR. ChIP was performed using sheared chromatin from 1 million cells. A titration consisting of 1, 2, 5 and 10 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoters of the active GAPDH and EIF4A2 genes, and for the inactive MYOD1 gene and the Sat2 satellite repeat. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

• WB

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Western blot - Anti-Histone H3 antibody [1B1B2] - Nuclear Marker and ChIP Grade (AB195277)

The antibody was diluted 1 : 1,000 in TBS-Tween containing 5% skimmed milk.

All lanes:

Western blot - Anti-Histone H3 antibody [1B1B2] - Nuclear Marker and ChIP Grade (ab195277) at 1/1000 dilution

Lane 1:

HeLa whole cell extract at 30 µg

Lane 2:

K562 whole cell extract at 30 µg

Lane 3:

MCF7 whole cell extract at 30 µg

Lane 4:

U2OS whole cell extract at 30 µg

Lane 5:

HepG2 whole cell extract at 30 µg

Lane 6:

Jurkat whole cell extract at 30 µg

Lane 7:

NIH 3T3 whole cell extract at 30 µg

Lane 8:

E14Tg2a Mouse ES whole cell extract at 30 µg

Predicted band size: 15 kDa

false

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [1B1B2] - Nuclear Marker and ChIP Grade (AB195277)

All lanes:

Western blot - Anti-Histone H3 antibody [1B1B2] - Nuclear Marker and ChIP Grade (ab195277) at 1/5000 dilution

Lane 1:

HeLa cell lysates at 40 µg

Lane 2:

Recombinant histone H3 at 1 µg

Lane 3:

Recombinant histone H4 at 1 µg

Predicted band size: 15 kDa

false