SNRPA1 Antibody

Knockdown Validated: SNRPA1 Antibody [NBP2-33447]

Knockdown Validated: SNRPA1 Antibody [NBP2-33447] - After indicated time periods of SNRPA1 knockdown, cells were collected and analyzed by western blotting. Forty-eight hours after SNRPA1 depletion, protein expression levels of RAD51 and Chk1 were already reduced. At 24h after siSNRPA1 transfection, gamma-H2AX levels transiently increased and then declined. Image collected and cropped by CiteAb from the following publication (nature.com/articles/oncsis201670), licensed under a CC-BY license.

Immunoprecipitation: SNRPA1 Antibody [NBP2-33447]

Immunoprecipitation: SNRPA1 Antibody [NBP2-33447] - Immunoprecipitation of SNRPA1 using NBP2-33447 or Rabbit IgG isotype control at a 1:100 dilution, followed by pulldown with Protein A magnetic beads. IP image submitted by a verified customer review.

Western Blot: SNRPA1 Antibody [NBP2-33447] -

Western Blot: SNRPA1 Antibody [NBP2-33447] - Recruitment kinetics of SNRPA1 & SF3A3 after laser microirradiation & SNRPA1-dependent recruitment of RAD51 & BRCA1 to laser tracks. (a) Image-based cytometry reveals increased gammaH2AX formation after 72 h SNRPA1 knockdown in U2OS cells. SNRPA1 knockdown effciency is shown in the bar graph. U2OS cells transiently (b) or stably (c) expressing SNRPA1-GFP were microirradiated & protein recruitment followed in real-time. For transcription & splicing inhibition cells were pretreated with DRB (50 μM) or SSA (100 nM) for at least 1 h before damage induction. Representative confocal images & recruitment kinetics are shown. (d) As in (b & c) but in cells stably expressing SF3A3-GFP. (e) Generation of R-loops at laser tracks visualized by HB-GFP recruitment. HB-GFP recruitment kinetics resemble SNRPA1-GFP. (f) Schematic of SNRPA1 deletion constructs. (g) Representative confocal images of recruitment of wild-type & mutated GFP-tagged SNRPA1 after laser microirradiation. (h) Quantification of recruitment, non-recruitment & displacement of GFP-tagged SNRPA1 fusions. Ten cells were analyzed per condition. (i) Confirmation of siRNA-mediated depletion of SNRPA1 after 48 & 72 h knockdown in cells used in (j). (j) Recruitment of RAD51 & BRCA1 in control & SNRPA1-depleted U2OS cells. U2OS cells were transfected with control or SNRPA1 siRNA & 48 or 72 h later microirradiated, fixed after indicated repair times & stained for gammaH2AX, RAD51 & BRCA1. Representative confocal images & percentage of cells displaying RAD51 & BRCA1 recruitment are shown. Scale bar 5 or 10 μM (i). Statistically significant differences were determined using Student's t-test, *P<0.05, **P<0.01. Error bars represent s.e.m. RFU (relative fluorescence units). Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/27991914), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: SNRPA1 Antibody [NBP2-33447] -

Western Blot: SNRPA1 Antibody [NBP2-33447] - SNRPA1 knockdown impairs BRCA1 & RAD51 accumulation at endonuclease (FokI) cleaved single DSB sites & radiation-induced DSB sites. (a) Visualization of endonuclease-mediated DSB induced by Shield 1 addition in 2-6-5 reporter cells with mCherry-LacI-FokI fusion protein expression. Cells were transfected with indicated siRNA, & 48 h later 4-OHT & Shield 1 were added for 5 h to induce mCherry-LacI-FokI expression. Cells were fixed & immunostained with indicated antibodies. In SNRPA1-depleted cells, BRCA1 & RAD51 accumulation to endonuclease cleaved single DSB sites were significantly reduced. (b) U2OS cells were transfected with control or SNRPA1 siRNA & after indicated time periods irradiated with 2 Gy. Two hours after irradiation, cells were fixed & immunostained. Quantification of RAD51 (>12 foci), gammaH2AX (>12 foci), BRCA1 (>12 foci) & RPA (>12 foci) positive cells. For each condition, more than 400 cells were analyzed. (c) After indicated time periods of SNRPA1 knockdown, cells were collected & analyzed by western blotting. Forty-eight hours after SNRPA1 depletion, protein expression levels of RAD51 & Chk1 were already reduced. At 24 h after siSNRPA1 transfection, gammaH2AX levels transiently increased & then declined. The error bars represent s.e.m. from three independent experiments (n=3). Statistically significant differences between cells treated with control or splicing factor siRNA were determined using Student's t-test, *P<0.05, **P<0.01. Scale bar is 10 μm. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/27991914), licensed under a CC-BY license. Not internally tested by Novus Biologicals.