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Epigenetics

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Histone extracts (15 \u03bcg) from HeLa cells were analysed by Western blot using H3K9me2 (9B3) Monoclonal Antibody (Cat. No. bsm-53016M) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.
HeLa cells were stained with H3K9me2 (9B3) Monoclonal Antibody (bsm-53016M). Cells were fixed with 4% formaldehyde for 10\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K9me2 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.
To test the specificity an ELISA was performed using a serial dilution H3K9me2 (9B3) Monoclonal Antibody (bsm-53016M). The wells were coated with peptides containing the unmodified H3K9 as well as the mono-, di- and trimethylated H3K9 and the dimethylated H3K4 and H3K27. Figure shows a high specificity of the antibody for the modification of interest.
ChIP assays were performed using HeLa cells, H3K9me2 (9B3) Monoclonal Antibody (bsm-53016M) and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 1.6 million cells. A titration of the antibody consisting of 1, 3 and 9 \u03bcg per ChIP experiment was analysed. IgG (5 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the promoter and the coding region of the GAPDH gene, and for the RPL10 and HBB promoters. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Catalog #1: bsm-53016M

Applications: WB, ELISA, IF(ICC), IF

Reactivity: Human

Conjugate: Unconjugated

HeLa cells were stained with H3K27ac (1A3) Monoclonal Antibody (bsm-53022M) and with DAPI. Cells were fixed with 4% formaldehyde for 10\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K27ac antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two staining is shown on the right.
To test the specificity an ELISA was performed using a serial dilution of H3K27ac (1A3) Monoclonal Antibody (bsm-53022M). The wells were coated with peptides containing the unmodified H3K27 region as well as the acetylated H3K27 and the acetylated H3K9. Figure shows a high specificity of the antibody for the peptide containing the modification of interest.
ChIP assays were performed using HeLa cells, H3K27ac (1A3) Monoclonal Antibody (bsm-53022M) and optimized PCR primer pairs for qPCR. ChIP was performed using sheared chromatin from 1 million cells. A titration consisting of 0.5, 1, 2 and 5 \u00b5g of antibody per ChIP experiment was analyzed. IgG (1\u00b5g\/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoters of the EIF4A2 and GAPDH genes, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Catalog #1: bsm-53022M

Applications: WB, ELISA, IF(ICC)

Reactivity: Human

Conjugate: Unconjugated

HeLa cells were stained with H3K36me3 (11D1) Monoclonal Antibody (bsm-53021M) and DAPI. Cells were fixed with methanol and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K36me3 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.
To test the specificity an ELISA was performed using a serial dilution of H3K36me3 (11D1) Monoclonal Antibody (bsm-53021M). The wells were coated with peptides containing unmodified H3K36 region as well as the mono-, di- and trimethylated H3K36 and the trimethylated H3K9. Figure shows a high specificity of the antibody for the peptide containing the modification of interest.
ChIP assays were performed using HeLa cells, H3K36me3 (11D1) Monoclonal Antibody (bsm-53021M) and optimized PCR primer pairs for qPCR. ChIP was performed using sheared chromatin from 1 million cells. A titration consisting of 0.5, 1, 2 and 5 \u03bcg of antibody per ChIP experiment was analyzed. IgG (1 \u03bcg\/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the coding regions of the active EIF2S3 and CCT5 genes, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Catalog #1: bsm-53021M

Applications: WB, ELISA, IF

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bsm-53000M

Applications: WB, IF(ICC)

Reactivity: Human

Conjugate: Unconjugated

Whole cell extracts (40 \u03bcg) from HeLa cells from an untransfected control (lane 1) and transfected with TBP siRNA (lane 2), and were analysed by Western blot using TBP Monoclonal Antibody (bsm-53003M) diluted 1:500 in TBSTween containing 5% skimmed milk, followed by secondary antibody.
ChIP assays were performed using U2OS cells, TBP (2C9) Monoclonal Antibody (bsm-53003M) and optimized primer sets for qPCR. Sheared chromatin from 1x10e6 cells and 4 \u03bcg of antibody were used per ChIP experiment. QPCR was performed with primers for the promoter of the c-fos and GAPDH genes, a region 0.5 and 1 kb upstream of the GAPDH promoter, and exon 2 of the myoglobin gene as a negative control. Figure shows the recovery (the relative amount of immunoprecipitated DNA compared to input DNA) and the occupancy (ratio +\/- control target). These results demonstrate the occupancy of both promoters by TBP.

Catalog #1: bsm-53003M

Applications: WB, ChIP-seq

Reactivity: Human

Conjugate: Unconjugated

Western blot analysis was performed on 100 fmol ER alpha (ERa) and ER beta (ERb1) recombinant protein with ER alpha (5G7) Monoclonal Antibody (bsm-53007M) at a concentration of 7 \u03bcg\/ ml. Figure shows that the antibody is specific for ER alpha isoform, whereas the ER beta isoform is not recognized.
COS-7 cells transiently over-expressing human ER\u03b1 (left) or ER\u00df1 (right) were labeled with the ER alpha (5G7) Monoclonal Antibody (bsm-53007M), used at a concentration of 15 \u03bcg\/ml, followed by a biotinylated secondary antibody and peroxidase-labeled avidin. Figure shows the specificity of the antibody for the ER alpha isoform.
ChIP assays was performed using MCF7 cells treated with the ER agonist estradiol for 3 hours prior to harvesting, ER alpha (5G7) Monoclonal Antibody (bsm-53007M), and optimized primer sets for qPCR. Sheared chromatin from 3 million cells and 5 \u03bcg of antibody were used per ChIP experiment. Recovery (%: ChIP\/input) and occupancy (x fold: +ve\/-ve) are shown. QPCR was performed with primers for the GREB1 promoter and for exon 2 of the myoglobin gene used as a negative control. Figure shows the recovery (the relative amount of immunoprecipitated DNA compared to input DNA) and the occupancy (ratio +\/- control target). These results demonstrate the occupancy of the GREB1 promoter by ERalpha.

Catalog #1: bsm-53007M

Applications: WB, IHC-P, IHC, dot-blot

Reactivity: Human, Monkey

Conjugate: Unconjugated

Catalog #1: bsm-53011M

Applications: WB

Reactivity: Human

Conjugate: Unconjugated

Western blot was performed on whole cell (25 \u03bcg, lane 1) and histone extracts (15 \u03bcg, lane 2) from HeLa cells, and on 1 \u03bcg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using H3K9un (7D7) Monoclonal Antibody (bsm-53024M). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.
HeLa cells were stained with H3K9un (7D7) Monoclonal Antibody (bsm-53024M) and with DAPI. Cells were fixed with 4% formaldehyde for 10\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K9un antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.
ChIP assays were performed using HeLa cells, H3K9un (7D7) Monoclonal Antibody (bsm-53024M) and optimized PCR primer sets for qPCR. ChIP was performed on sheared chromatin from 1 million cells, using the SX-8G IP-Star automated system. A titration of the antibody consisting of 1, 2, 5, and 10 \u03bcg per ChIP experiment was analysed. IgG (2 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the coding regions of the MYOD1 and MB genes and for a region 1 kb upstream of the GAPDH promoter, used as positive controls, and for the ZNF510 coding region, the EIF4A2 promoter and the Sat2 satellite repeat region, used as negative controls. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Catalog #1: bsm-53024M

Applications: WB, IHC-P, IF(ICC)

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bsm-53025M

Applications: WB, IF(ICC)

Reactivity: Human

Conjugate: Unconjugated

Whole cell extracts from HL60 and NB4 cells (40 \u03bcg) were analysed by Western blot using TET2 (6A1) Monoclonal Antibody (bsm-53017M), diluted 1:2,000 in PBS containing 10% milk. The position of the protein of interest (expected MW 224 kDa) is indicated on the right; the marker (in kDa) is shown on the left.
IP was performed on 250 \u03bcg HL60 RIPA cell lysate using TET2 (6A1) Monoclonal Antibody (bsm-53017M) (lane 3) or an IgG negative control (lane 2). The input sample (25 \u03bcg RIPA lysate) was used as a positive control (lane 1). The samples were analysed by Western blot using TET2 (6A1) Monoclonal Antibody (bsm-53017M), diluted 1:2,000 in PBS containing 10% milk.

Catalog #1: bsm-53017M

Applications: WB, IP

Reactivity: Human, Mouse

Conjugate: Unconjugated

Catalog #1: bsm-53018M

Applications: WB

Reactivity: Human, Mouse

Conjugate: Unconjugated