Cells were lysed with SDS sample buffer (50 mM Tris at pH 7.4, 100 mM NaCl, 1% Triton X-100, 0.1% SDS, 1% sodium deoxycholate, 1× proteinase inhibitor cocktail [Biotools]), resolved by SDS-PAGE, transferred to nitrocellulose membranes, and incubated with primary antibodies detecting CDH1 (1:1000; Cell Signaling), hnRNP H1 (1:1000; Bethyl Laboratories), hnRNP F (1:1500; Santa Cruz Biotechnology), GAPDH (1:20,000; Sigma), or ACTB (1:30,000; Sigma) overnight at 4°C. Secondary antibodies HRP-conjugated anti-mouse and anti-rabbit IgG (Sigma) were used at 1:10,000 for 2 h at room temperature. Following washes with PBST (1× PBS, 0.1% Tween-20), ECL-Prime (GE Healthcare) was used to visualize the chemiluminescence signal, and ImageJ software was used for quantification.
Hnrnp h1
Manufactured by Fortis Life Sciences
HnRNP H1 is a protein that is involved in the regulation of RNA processing and gene expression. It is a member of the heterogeneous nuclear ribonucleoprotein (hnRNP) family of proteins. HnRNP H1 plays a role in the splicing and stability of mRNA molecules.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using hnrnp h1
1
Western Blot Analysis of Cellular Proteins
2
Western Blot Analysis of RNA-Binding Proteins
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Cells were lysed with SDS sample buffer (50 mM Tris pH 7.4, 100 mM NaCl, 1% Triton X-100, 0.1% SDS, 1% sodium deoxycholate, and 1× proteinase inhibitor cocktail [Biotools]) and resolved by SDS-PAGE. Proteins were then transferred to nitrocellulose membranes and incubated with primary antibodies detecting PTBP1 (BB7) (Sigma, 1:8000), DDX21 (Bethyl Laboratories, 1:1000), hnRNPH1 (Bethyl Laboratories, 1:1500), or ACTB (Sigma, 1:30,000) overnight at 4°C. Secondary HRP-conjugated anti-mouse and anti-rabbit IgGs (Sigma) were used at 1:10,000 for 2 h at room temperature. Following washes with PBST (1× PBS, 0.1% Tween-20), ECL-Prime (GE Healthcare) was used to visualize the chemiluminescence signal, and Image J software was used for quantification.
3
HSATIII RNA In Situ Hybridization and Immunostaining
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For HSATIII RNA in situ hybridization, cells cultured on cover glasses were washed with ice-cold PBS and fixed in 4% PFA/PBS for 10 min at room temperature. The cover glasses were then washed twice with ice-cold PBS and permeabilized in cold 70% ethanol for at least 1 hour at 4°C. Bethyl Laboratories, A300-790A; SLTM: Bethyl Laboratories, A302-834A; HNRNPA1: MBL, RN114PW; HNRNPH1: Bethyl Laboratories, IHC-00087). After washing in TBST, the cover glasses were incubated in 3% BSA/TBST containing Alexa 488-or Alexa 568-conjugated secondary antibodies. For immunostaining, the fixed cells were permeabilized in cold PBS containing 0.1% Triton X-100 for 15 min, and then incubated with the antibodies as described above. For the triple staining shown in Figure 1C, HSATIII was visualized with a biotin-labeled HSATIII ASO and avidin DN-FITC (Vector Laboratories). Primary antibodies against the proteins of interest were visualized with Alexa 350-or Alexa 568-conjugated secondary antibodies
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