Anti-n-Myc/MYCN antibody [EPR18982-8R-3] - ChIP Grade

• IP

Unknown

Immunoprecipitation - Anti-n-Myc/MYCN antibody [EPR18982-8R-3] - ChIP Grade (AB227822)

n-Myc/MYCN was immunoprecipitated from 0.35 mg of IMR-32 (human neuroblastoma neuroblast cell line) whole cell lysate with ab227822 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab227822 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1 : IMR-32 whole cell lysate 10 μg (Input).

Lane 2 : ab227822 IP in IMR-32 whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab227822 in IMR-32 whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 3 seconds.

The expression profile observed is consistent with what has been described in the literature (PMID : 17938259; PMID : 2657399).

All lanes:

Immunoprecipitation - Anti-n-Myc/MYCN antibody [EPR18982-8R-3] - ChIP Grade (ab227822)

Predicted band size: 50 kDa

Observed band size: 49-62 kDa

false

• ChIP

Unknown

ChIP - Anti-n-Myc/MYCN antibody [EPR18982-8R-3] - ChIP Grade (AB227822)

Chromatin was prepared from IMR-32 cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The ChIP was performed with 25 μg of chromatin, 5 μg of ab227822 (red), and 20 μl of Protein A/G sepharose beads. 5 μg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (SYBR green approach). Primers and probes are located in the first kb of the transcribed region.

• WB

Unknown

Western blot - Anti-n-Myc/MYCN antibody [EPR18982-8R-3] - ChIP Grade (AB227822)

Blocking and dilution buffer : 5% NFDM/TBST

The expression profile observed is consistent with what has been described in the literature (PMID : 11034201; PMID : 27197171; PMID : 23792191).

Negative control : HeLa (PMID : 27197171).

All lanes:

Western blot - Anti-n-Myc/MYCN antibody [EPR18982-8R-3] - ChIP Grade (ab227822) at 1/1000 dilution

Lane 1:

IMR-32 (human neuroblastoma neuroblast cell line) whole cell lysate at 20 µg

Lane 2:

HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 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 1/100000 dilution

Predicted band size: 50 kDa

Observed band size: 62 kDa

false

Exposure time: 103s