Blocking Assay:Article Title: Intermolecular 3′UTR-3′UTR interactions drive Wnt gene activation through heteromeric protein assembly
Article Snippet: Total protein was collected with Laemmli buffer and boiled at 95°C for 10 min. Proteins were resolved by NuPAGETM Bis-Tris Mini Protein Gels (Invitrogen) and transferred to a nitrocellulose membrane. .. Membranes were blocked in Intercept® Blocking Buffer (Li-COR) and incubated with primary antibodies overnight at 4°C. ..
Article Title: Ecological determinants of disease and immunity in myelodysplastic syndromes
Article Snippet: Protein concentration was measured using the BCA Protein Assay Kit (Pierce, #23225). .. Protein lysates were resolved on 4–12% Bis-Tris gels (Invitrogen, NP0335BOX), transferred to nitrocellulose membranes using the iBlot 2 Gel Transfer Device (Invitrogen, #IB21001), and blocked with Intercept Blocking Buffer (LI-COR, #927-60003). .. The following primary antibodies were used: TGFBR1 (Abcam, #ab235578), GAPDH (Cell Signaling Technology, #97166), U2AF2 (Cell Signaling Technology, #70471), CD98/SLC3A2 (1:5000; Cell Signaling Technology, #47213), β-actin (1:5000; Cell Signaling Technology, #3700).
Article Title: An interaction between HP1 and the Chromosomal Passenger Complex Initiates Acentrosomal Spindle Assembly in Drosophila oocytes
Article Snippet: 1 mg of total protein was loaded per lane on SDS-PAGE gels and transferred to a nitrocellulose membrane (Bio-Rad) using a Trans-Blot Turbo machine (Bio-Rad). .. Membranes were blocked with Intercept Blocking Buffer (LI-COR) for 1h at room temperature, then sequentially incubated with primary and secondary antibodies from LI-COR (Supplementary Table 1). .. Blots were imaged using the Odyssey CLX Imaging System (LI-COR Biosciences).
Article Title: Androgen loss accelerates brain tumour growth via HPA axis activation.
Article Snippet: A total of 20–25 μg protein lysate was loaded onto 4–20% Tris Glycine gels (Thermo Fisher) and proteins were transferred to nitrocellulose membranes. .. After blocking with Intercept Blocking buffer (LiCor Biosciences), membranes were incubated with primary mouse anti-caspase-1 antibody (1:1,000; Adipogen), followed by IR-680-conjugated anti-mouse secondary antibody (1:10,000; LiCor Bio). .. In brief, after live/dead staining with LIVEDEAD Blue (Thermo Fisher Scientific) on ice for 15 min, cells were washed and incubated with FcR blocker (Miltenyi Biotech) diluted in PBS and 2% BSA on ice for 10 min. For surface staining, cells were incubated in an antibody mixture diluted in brilliant buffer (BD Biosciences) at 1:100 to 1:250 on ice for 30 min. After washing with PBS and 2% BSA buffer, cells were fixed with FOXP3/ transcription factor fixation buffer (eBioscience) at 4 °C overnight.
Article Title: Extracellular matrix microarchitecture modulates cellular behavior and extracellular vesicle phenotypes in biomimetic tendon models
Article Snippet: Following electrophoresis, proteins were transferred onto PVDF membranes using a semi-dry blotting system (Bio-Rad; Cat. No. 1704156). .. Membranes were blocked at room temperature for 1 h using Intercept® Blocking Buffer (LI-COR; Cat. No. 927-70001) to minimize nonspecific binding. ..
Article Title: Adaptive loss of function accelerated the evolution of ancient and modern human cognition
Article Snippet: The membrane was blocked using Intercept Blocking Buffer (cat no. 927-60001, LI-COR Biotechnology, Lincoln, NE) (60 rpm, 1 h, room temperature) and then incubated in the appropriate primary antibody mixture (4°C, overnight). .. The membrane was blocked using Intercept Blocking Buffer (cat no. 927-60001, LI-COR Biotechnology, Lincoln, NE) (60 rpm, 1 h, room temperature) and then incubated in the appropriate primary antibody mixture (4°C, overnight). .. Primary antibodies used were: α-ESR1 (cat no. 8644S, Cell Signaling Technology, Danvers, MA; 1:1000 dilution), α-mCherry (cat no. 43590S, Cell Signaling Technology, 1:1000 dilution), α-DYKDDDDK tag (cat no. 14793S, Cell Signaling Technology, 1:1000 dilution) for EYA1, AKAP11, and TSHR, α-RET (cat no. 14556, Cell Signaling Technology, 1:1000), α-GAPDH (cat no. 97166S, Cell Signaling Technology, 1:5,000 dilution), α-beta-actin (cat no. 3700S, Cell Signaling Technology, 1:5000 dilution) and α-alpha-tubulin (cat no. MS581P1, Millipore Sigma, 1:5000 dilution).
Article Title: Disruption of the LRRK2 substrate RAB12 facilitates neurotransmission and causes hyperactivity in mice.
Article Snippet: .. Membranes were blocked with Intercept® Blocking Buffer (LI-COR, #927-60001) at room temperature for one hour, followed by overnight incubation at 4 °C with primary antibodies: rabbit anti-pRAB12 (Abcam, #ab256487, 1:1,000), mouse antiRAB12 (Santa Cruz Biotechnology, #sc-515613, 1:1,000), mouse anti-β-Actin (Cell Signaling Technology, #3700S, 1:10,000), mouse anti-synaptophysin (Synaptic Systems, #101011, 1:1,000), rat anti-GFAP (Invitrogen, #13-0300, 1:1,000), rabbit anti-RILPL1 (BOSTER, #A13355, 1:1,000), rabbit anti-β-Actin (Cell Signaling Technology, #4970S, 1:10,000), mouse anti-PSD95 (Millipore, #MAB1598, 1: 1000), rabbit anti-SV2A (SYSY, #119002, 1: 3000), Guinea pig anti-vGLUT1(SYSY, #135304, 1:1000), rabbit anti-CLSTN3 (Proteintech, #13302-1- AP, 1:800), rabbit anti-RPH3A (Proteintech, #11396-1-AP, 1:2000), rabbit anti-synaptotagmin-1 (CST, #3347S), and rabbit anti-SNAP25 (SYSY, #111002, 1:5000). .. Following primary antibody incubation, the membranes were washed three times with TBST, five minutes each, and incubated with appropriate HRP-conjugated secondary antibodies (Cell Signaling Technology, #7074S for pRAB12, RILPL1, SV2A, CLSTN3, RPH3A, synaptotagmin-1, and SNAP25; #7076S for total RAB12, synaptophysin, PSD95, and mouse anti-β-Actin; Santa Cruz Biotechnology, #sc-2006 for GFAP) for one hour at room temperature.
Article Title: Insulin Receptor Isoform Pattern in Aged Normal and Alzheimer's Human Cortex
Article Snippet: The protein was transferred onto methanol‐wetted polyvinylidene fluoride (PVDF) membranes (cat. no. GE10600023, Amersham Biosciences) for 90 min at 100 V in a Mini Trans‐Blot Cell module (Bio‐Rad) with ice‐cold NuPAGE transfer buffer (cat. no. NP0006, Invitrogen) containing 20% methanol. .. After protein transfer, the membranes were incubated for an hour with Intercept blocking buffer (cat. no. 927‐70001, LICORbio). .. Each membrane was incubated with a mouse anti‐β‐actin antibody as a loading control (cat. no. ab6276, Abcam; 1:1000) and one of three rabbit anti‐IRβ antibodies (Novus Biologicals, NBP2‐12793, 1:500; Invitrogen, PA5‐27334, 1:500; or Abbiotec, 250 724, 1:300) in a solution of 5% bovine serum albumin (BSA) in tris‐buffered saline with 0.1% Tween 20 (TBS‐T) at 4°C overnight with agitation.
Incubation:Article Title: Intermolecular 3′UTR-3′UTR interactions drive Wnt gene activation through heteromeric protein assembly
Article Snippet: Total protein was collected with Laemmli buffer and boiled at 95°C for 10 min. Proteins were resolved by NuPAGETM Bis-Tris Mini Protein Gels (Invitrogen) and transferred to a nitrocellulose membrane. .. Membranes were blocked in Intercept® Blocking Buffer (Li-COR) and incubated with primary antibodies overnight at 4°C. ..
Article Title: An interaction between HP1 and the Chromosomal Passenger Complex Initiates Acentrosomal Spindle Assembly in Drosophila oocytes
Article Snippet: 1 mg of total protein was loaded per lane on SDS-PAGE gels and transferred to a nitrocellulose membrane (Bio-Rad) using a Trans-Blot Turbo machine (Bio-Rad). .. Membranes were blocked with Intercept Blocking Buffer (LI-COR) for 1h at room temperature, then sequentially incubated with primary and secondary antibodies from LI-COR (Supplementary Table 1). .. Blots were imaged using the Odyssey CLX Imaging System (LI-COR Biosciences).
Article Title: Androgen loss accelerates brain tumour growth via HPA axis activation.
Article Snippet: A total of 20–25 μg protein lysate was loaded onto 4–20% Tris Glycine gels (Thermo Fisher) and proteins were transferred to nitrocellulose membranes. .. After blocking with Intercept Blocking buffer (LiCor Biosciences), membranes were incubated with primary mouse anti-caspase-1 antibody (1:1,000; Adipogen), followed by IR-680-conjugated anti-mouse secondary antibody (1:10,000; LiCor Bio). .. In brief, after live/dead staining with LIVEDEAD Blue (Thermo Fisher Scientific) on ice for 15 min, cells were washed and incubated with FcR blocker (Miltenyi Biotech) diluted in PBS and 2% BSA on ice for 10 min. For surface staining, cells were incubated in an antibody mixture diluted in brilliant buffer (BD Biosciences) at 1:100 to 1:250 on ice for 30 min. After washing with PBS and 2% BSA buffer, cells were fixed with FOXP3/ transcription factor fixation buffer (eBioscience) at 4 °C overnight.
Article Title: Adaptive loss of function accelerated the evolution of ancient and modern human cognition
Article Snippet: The membrane was blocked using Intercept Blocking Buffer (cat no. 927-60001, LI-COR Biotechnology, Lincoln, NE) (60 rpm, 1 h, room temperature) and then incubated in the appropriate primary antibody mixture (4°C, overnight). .. The membrane was blocked using Intercept Blocking Buffer (cat no. 927-60001, LI-COR Biotechnology, Lincoln, NE) (60 rpm, 1 h, room temperature) and then incubated in the appropriate primary antibody mixture (4°C, overnight). .. Primary antibodies used were: α-ESR1 (cat no. 8644S, Cell Signaling Technology, Danvers, MA; 1:1000 dilution), α-mCherry (cat no. 43590S, Cell Signaling Technology, 1:1000 dilution), α-DYKDDDDK tag (cat no. 14793S, Cell Signaling Technology, 1:1000 dilution) for EYA1, AKAP11, and TSHR, α-RET (cat no. 14556, Cell Signaling Technology, 1:1000), α-GAPDH (cat no. 97166S, Cell Signaling Technology, 1:5,000 dilution), α-beta-actin (cat no. 3700S, Cell Signaling Technology, 1:5000 dilution) and α-alpha-tubulin (cat no. MS581P1, Millipore Sigma, 1:5000 dilution).
Article Title: Disruption of the LRRK2 substrate RAB12 facilitates neurotransmission and causes hyperactivity in mice.
Article Snippet: .. Membranes were blocked with Intercept® Blocking Buffer (LI-COR, #927-60001) at room temperature for one hour, followed by overnight incubation at 4 °C with primary antibodies: rabbit anti-pRAB12 (Abcam, #ab256487, 1:1,000), mouse antiRAB12 (Santa Cruz Biotechnology, #sc-515613, 1:1,000), mouse anti-β-Actin (Cell Signaling Technology, #3700S, 1:10,000), mouse anti-synaptophysin (Synaptic Systems, #101011, 1:1,000), rat anti-GFAP (Invitrogen, #13-0300, 1:1,000), rabbit anti-RILPL1 (BOSTER, #A13355, 1:1,000), rabbit anti-β-Actin (Cell Signaling Technology, #4970S, 1:10,000), mouse anti-PSD95 (Millipore, #MAB1598, 1: 1000), rabbit anti-SV2A (SYSY, #119002, 1: 3000), Guinea pig anti-vGLUT1(SYSY, #135304, 1:1000), rabbit anti-CLSTN3 (Proteintech, #13302-1- AP, 1:800), rabbit anti-RPH3A (Proteintech, #11396-1-AP, 1:2000), rabbit anti-synaptotagmin-1 (CST, #3347S), and rabbit anti-SNAP25 (SYSY, #111002, 1:5000). .. Following primary antibody incubation, the membranes were washed three times with TBST, five minutes each, and incubated with appropriate HRP-conjugated secondary antibodies (Cell Signaling Technology, #7074S for pRAB12, RILPL1, SV2A, CLSTN3, RPH3A, synaptotagmin-1, and SNAP25; #7076S for total RAB12, synaptophysin, PSD95, and mouse anti-β-Actin; Santa Cruz Biotechnology, #sc-2006 for GFAP) for one hour at room temperature.
Article Title: Insulin Receptor Isoform Pattern in Aged Normal and Alzheimer's Human Cortex
Article Snippet: The protein was transferred onto methanol‐wetted polyvinylidene fluoride (PVDF) membranes (cat. no. GE10600023, Amersham Biosciences) for 90 min at 100 V in a Mini Trans‐Blot Cell module (Bio‐Rad) with ice‐cold NuPAGE transfer buffer (cat. no. NP0006, Invitrogen) containing 20% methanol. .. After protein transfer, the membranes were incubated for an hour with Intercept blocking buffer (cat. no. 927‐70001, LICORbio). .. Each membrane was incubated with a mouse anti‐β‐actin antibody as a loading control (cat. no. ab6276, Abcam; 1:1000) and one of three rabbit anti‐IRβ antibodies (Novus Biologicals, NBP2‐12793, 1:500; Invitrogen, PA5‐27334, 1:500; or Abbiotec, 250 724, 1:300) in a solution of 5% bovine serum albumin (BSA) in tris‐buffered saline with 0.1% Tween 20 (TBS‐T) at 4°C overnight with agitation.
Binding Assay:Article Title: Extracellular matrix microarchitecture modulates cellular behavior and extracellular vesicle phenotypes in biomimetic tendon models
Article Snippet: Following electrophoresis, proteins were transferred onto PVDF membranes using a semi-dry blotting system (Bio-Rad; Cat. No. 1704156). .. Membranes were blocked at room temperature for 1 h using Intercept® Blocking Buffer (LI-COR; Cat. No. 927-70001) to minimize nonspecific binding. ..
Membrane:Article Title: Adaptive loss of function accelerated the evolution of ancient and modern human cognition
Article Snippet: The membrane was blocked using Intercept Blocking Buffer (cat no. 927-60001, LI-COR Biotechnology, Lincoln, NE) (60 rpm, 1 h, room temperature) and then incubated in the appropriate primary antibody mixture (4°C, overnight). .. The membrane was blocked using Intercept Blocking Buffer (cat no. 927-60001, LI-COR Biotechnology, Lincoln, NE) (60 rpm, 1 h, room temperature) and then incubated in the appropriate primary antibody mixture (4°C, overnight). .. Primary antibodies used were: α-ESR1 (cat no. 8644S, Cell Signaling Technology, Danvers, MA; 1:1000 dilution), α-mCherry (cat no. 43590S, Cell Signaling Technology, 1:1000 dilution), α-DYKDDDDK tag (cat no. 14793S, Cell Signaling Technology, 1:1000 dilution) for EYA1, AKAP11, and TSHR, α-RET (cat no. 14556, Cell Signaling Technology, 1:1000), α-GAPDH (cat no. 97166S, Cell Signaling Technology, 1:5,000 dilution), α-beta-actin (cat no. 3700S, Cell Signaling Technology, 1:5000 dilution) and α-alpha-tubulin (cat no. MS581P1, Millipore Sigma, 1:5000 dilution).
|
LI-COR is a verified supplier 
LI-COR manufactures this product 