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20 Full Site Results for "lysates": Protocol
Immunoprecipitation Protocol (NB200-174)
Immunoprecipitation 1. Total cell lysates (0.25 mg) from NIH3T3 cells which have deleted the Arf gene and from NIH3T3 cells expressing an HA-tagged version of p19Arf were incubated with control rat IgG or anti-p19Arf (NB 200-174) and protein G- Seph
Western Blot Protocol Specific for Survivin Antibody (NB500-201)
Western Blot Procedure 1) Cells were pelleted, washed in 1XPBS, suspended in ice water (~ 5 x 10(6) cells/ml), and placed on ice 2) Lysates were prepared with the addition of 2X lysis buffer [2% SDS/ 50mM Tris-HCl / 10% glycerol] 3) Lysates were heat
Western Blot protocol for Bestrophin 1 Antibody (NB300-164)
Procedure Guide for NB 300-164 Monoclonal Anti-Bestrophin Western Blot Procedure 1. Run cell lysates** on an SDS-PAGE gel. 2. Transfer the proteins to PVDF. 3. Block the membrane in 1% Carnation instant milk in PBS + 0.1% Tween 20 (with 0.1mM CaCl2
Western Blot Protocols specific for 15-PGDH Antibody (NB200-179)
Western Blot I (LoVo lysates) 1. Perform SDS-PAGE (4-12%) on samples to be analyzed, loading 20ug of total protein per lane. 2. Transfer proteins to Nitrocellulose according to the instructions provided by the manufacturer of the transferapparatu
Western Blot Protocol for PPM1B Antibody (NBP1-04335)
Western Blot: 1. Sample preparation: SDS loading buffer was added to MCF7 cell lysates and recombinant PPM1B protein. The solution was boiled before loading on a 15% SDS-PAGE 2. Blocking: 5% skim milk in PBS-Tween, for 1hr at room temperature, 3 tim
Western Blot Protocol for GABA B Receptor 1 Antibody (NB300-145)
Procedure Guide for NB 300-145 Polyclonal Anti-GABA_R1 Western Blot Procedure 1) Resolve ~90 mg of whole brain or cerebral cortex tissue lysates on a 4-20% Tris-Glycine gel. 2) Transfer to nitrocellulose membranes. 3) Block membranes for 1 hour, at
Protocol specific for Survivin Antibody (NB500-205)
Western Blot Procedure 1) HeLa whole cell lysates were heated to 70 degrees C for 10 minutes and then microfuged at room temperature. 2) 65 mg of lysate were electrophoresed (200 V) through a 4-12% NuPAGE Bis-Tris gel, for 1 hr. 3) Proteins were
WB protocol specific for Fatty Acid Synthase Antibody (NB400-114)
1. Run ~50 ug of total protein on a 4-15% SDS polyacrylamide gel. 2. Transfer protein to a nitrocellulose membrane. 3. Block membrane with 1XPBS/5% non-fat milk/0.1% Tween-20 for 1 hour at room temperature (~23-27C). 4. Incubate membrane with 1:1000
Immunoprecipitation protocol specific for Beclin 1 Antibody (NB500-249)
Immunoprecipitation Protocol: 1. Cells in 2x 75cm flasks (60% confluency) are scraped with 0.5ml of Tris lysis Buffer (50mM Tris, 150mM NaCl, 1mM EDTA, 100ug/ml PMSF, 1% triton). 2. Lyse 1h at 4C, with gentle agitation. 3. Centrifuge to clear the
Western Blot Protocol for HIF-1 alpha Antibody (NB100-105)
Western Blot Protocol Specific for HIF-1alpha Antibody (NB100-105) General considerations for Western blot analysis of HIF-alpha proteins 1. HIF-1alpha is largely undetectable in cells or tissues grown under normoxic conditions. It is stabilized on
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