All lanes : Anti-PARP1 antibody (ab227244) at 1/5000 dilution

Lane 1 : Non-transfected HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell extract
Lane 2 : PARP1 shRNA transfected HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell extract

Lysates/proteins at 30 µg per lane.

Secondary
All lanes : HRP-conjugated anti-rabbit IgG

Predicted band size: 113 kDa

7.5% SDS-PAGE gel.

4% paraformaldehyde-fixed HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained for PARP1 (green) using ab227244 at 1/500 dilution in ICC/IF.

Red: Phalloidin, a cytoskeleton marker, at 1/200 dilution.

Blue: Hoechst 33342 staining.

ChIP was performed with HeLa chromatin extract and 5 µg of either normal rabbit IgG or ab227244. The precipitated DNA was detected by PCR with primer set targeting to HSP70.1 promoter.

Paraffin-embedded HeLa xenograft tissue stained for PARP1 using ab227244 at 1/500 dilution in immunohistochemical analysis.

Antigen Retrieval: EDTA based, pH 8.0, buffer, 15minutes. 

PARP1 was immunoprecipitated from HCT 116 (human colorectal carcinoma cell line) whole cell extract with 4 µg ab227244. Western blot was performed from the immunoprecipitate using ab227244 at 1/500 dilution. Anti-Rabbit IgG was used as a secondary reagent.

Lane 1: HCT 116 whole cell extract 30 μg.

Lane 2: Control IP in HCT 116 whole cell extract with 4 μg of preimmune Rabbit IgG.

Lane 3: ab227244 IP in HCT 116 whole cell extract.

All lanes : Anti-PARP1 antibody (ab227244) at 1/1000 dilution

Lane 1 : 293T whole cell extracts
Lane 2 : A431 whole cell extracts
Lane 3 : HeLa whole cell extracts
Lane 4 : HepG2 whole cell extracts

Lysates/proteins at 30 µg per lane.

Secondary
All lanes : Rabbit IgG antibody (HRP) at 1/10000 dilution

Predicted band size: 113 kDa

5% gel.

Running conditions: 80V, 15min; 140V, 40 minutes.

Transfer condition: Semi-dry, 18 V, 60 min (NC membrane).

Blocking condition: 5% non-fat milk in TBST, RT, 60 minutes.

Primary antibody incubation: 4? , overnight.

Washing condition: 5 ml TBST, 4 x 5 minutes. 

Exposure: chemiluminescent substrate for the detection of HRP-conjugated antibody

 

All lanes : Anti-PARP1 antibody (ab227244) at 1/1000 dilution

All lanes : Neuro2A whole cell extracts

Lysates/proteins at 30 µg per lane.

Secondary
All lanes : Rabbit IgG antibody (HRP) at 1/10000 dilution

Predicted band size: 113 kDa

5% gel.

Running conditions: 80V, 15min; 140V, 40 minutes.

Transfer condition: Semi-dry, 18 V, 60 min (NC membrane).

Blocking condition: 5% non-fat milk in TBST, RT, 60 minutes.

Primary antibody incubation: 4? , overnight.

Washing condition: 5 ml TBST, 4 x 5 minutes. 

Exposure: chemiluminescent substrate for the detection of HRP-conjugated antibody

 

All lanes : Anti-PARP1 antibody (ab227244) at 1/5000 dilution

Lane 1 : HCT116 whole cell lysate (untreated)
Lane 2 : HCT116 whole cell lysate (30 µM cisplatin treatment for 24 hours)

Lysates/proteins at 30 µg per lane.

Secondary
All lanes : HRP-conjugated anti-rabbit IgG

Predicted band size: 113 kDa

7.5% SDS-PAGE gel.

The top band is full-length and the bottom band on lane 2 is cleaved PARP1. 

Cross-linked ChIP was performed with Raji chromatin extract and 5 μg of either control rabbit IgG or ab227244. The precipitated DNA was detected by PCR with primer set targeting to S100A9 promoter.

All lanes : Anti-PARP1 antibody (ab227244) at 1/500 dilution

Lane 1 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell extract
Lane 2 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell extract

Secondary
All lanes : HRP-conjugated anti-rabbit IgG

Predicted band size: 113 kDa

7.5% SDS-PAGE gel.