Anti-SNF2H 抗体 - ChIP Grade

• ChIP

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ChIP - Anti-SNF2H antibody - ChIP Grade (AB3749)

ab3749 works in ChIP, as shown by the detection of an increase in the recruitment of SNF2H to the estrogen-responsive pS2 promoter. Sonicated Chromatin prepared from untreated or 17beta-estradiol (E) treated MCF7 cells was subjected to the ChIP procedure with ab3749 antibody specific to SNF2H. The immunoprecipitated chromatin was analysed in the proximal region of the estro

• WB

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Western blot - Anti-SNF2H antibody - ChIP Grade (AB3749)

Rabbit polyclonal ab3749 to SNF2H was used in all lanes at 1/500 dilution.

Lane 1 : HeLa lysate (20ug)
Lane 2 : Mouse 3T3 lysate (20ug)
Lane 3 : HeLa lysate (20ug), blocking peptide (ab5160) 1.0ug
Lane 4 : Mouse 3T3 lysate (20ug), blocking peptide (ab5160) 1.0ug

Secondary antibody : Goat polyclonal to Rabbit IgG (ab6721)

ab3749 was able to recognise SNF2H in HeLA and Mouse 3T3 lysates (band at ~120 kDa). Binding was blocked by the immunising peptide ab5160.

Rabbit polyclonal ab3749 to SNF2H was used in all lanes at 1/500 dilution.

Lane 1 : HeLa lysate (20ug)
Lane 2 : Mouse 3T3 lysate (20ug)
Lane 3 : HeLa lysate (20ug), blocking peptide (ab5160) 1.0ug
Lane 4 : Mouse 3T3 lysate (20ug), blocking peptide (ab5160) 1.0ug

Secondary antibody : Goat polyclonal to Rabbit IgG (ab6721)

ab3749 was able to recognise SNF2H in HeLA and Mouse 3T3 lysates (band at ~120 kDa). Binding was blocked by the immun

All lanes:

Western blot - Anti-SNF2H antibody - ChIP Grade (ab3749)

Predicted band size: 122 kDa

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• ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-SNF2H antibody - ChIP Grade (AB3749)

Immunocytochemistry-immunofluorescence using Anti-SNF2H antibody - ChIP Grade, ab3749. Publication image from Nishitani, H. et al., 2017, Nat Commun, 28719581. Legend direct from paper.

The ACF1–SNF2H chromatin remodeller interacts with HBO1 and accumulates at ultraviolet-damaged DNA.(a) ShCtl, shHBO1 and shDDB2 cells were irradiated with 50 J m−2 ultraviolet and cultured for 30 min. Cells were co-immunostained with indicated antibodies. Scale bars, 5 µm. (b) Depletion of HBO1-suppressed ACF1 accumulation at damage sites. ShCtl and shHBO1 cells were irradiated with 50 J m−2 ultraviolet and cultured for 30 min. Relative ACF1/DDB2 intensities of shHBO1 cells are shown as % of shCtl cells. Cells (n=50) from three independent experiments; error bars indicate means±s.d. **P<0.01 (Student’s t-test). (c) Live-cell imaging of GFP-SNF2H accumulation at damage sites. GFP-SNF2H was transiently transfected in shCtl, shDDB2 and shHBO1 cells. Laser irradiation was performed as described in Methods. Cells (n=30) at least from three independent experiments; error bars are the mean±s.e.m. *P<0.05, **P<0.01 (Student’s t-test). (d) HEK293 cells overexpressing GFP-SNF2H and HA-DDB2 or Myc-HBO1 were immunoprecipitated with anti-HA antibody, anti-Myc-antibody and anti-GFP agarose. Immunoprecipitated samples were subjected to western blotting with the indicated antibodies. (e) Lysates from ultraviolet-irradiated or non-irradiated HeLa cells were immunoprecipitated with anti-SNF2H antibody. Immunoprecipitates were subjected to western blotting with the indicated antibodies. (f) Live-cell imaging of XPC–EGFP accumulation at damage sites. XPC–EGFP and siCtl or siSNF2H were transiently transfected in shCtl and shHBO1 cells. Laser irradiation was performed as described in Methods. Cells (n=30) from three independent experiments; error bars are the mean±s.e.m. **P<0.01 (Student’s t-test). (g) SiCtl or siSNF2H was transiently transfected in shCtl and shHBO1 cells. Cells were irradiated at indicated doses of ultraviolet. The numbers of colonies were counted and are represented as % survival of mock-irradiated cells. Three independent experiments were performed. Error bars indicate means±s.d. (h) Histone H3 Lys 4 tri-methylation was rapidly decreased in shHBO1 cells after ultraviolet. ShCtl and shHBO1 cells were irradiated with 40 J m−2 ultraviolet and collected at indicated times. Chromatin-enriched fractions were subjected to western blotting using the indicated antibodies.