seeblue protein ladder Ctbp1 western blot

#SEEBLUE PROTEIN LADDER CTBP1 WESTERN BLOT PLUS#

Chloride (–) is from the gel buffer and serves as a leading ion due to its high affinity to the anode relative to other anions in the system.Bis-Tris Gels use a Bis-Tris discontinuous buffer system involving three ions:

The gels can be run using MES SDS running buffer to better resolve smaller proteins (1–200 kDa) and MOPS SDS running buffer to resolve medium- to large-size proteins (14–260 kDa). For optimal sample preparation, we recommend using buffers containing SDS.

#SEEBLUE PROTEIN LADDER CTBP1 WESTERN BLOT PLUS#

NuPAGE Bis-Tris and Bolt Bis-Tris Plus Gels are formulated for denaturing gel electrophoresis applications. High capacity wells of Bolt Bis-Tris Plus Gels-wedge-shaped wells makes sample loading easier (no specialized pipette tips required) and allow loads of up to two times higher sample volume.Longer shelf life-NuPAGE Bis-Tris and Bolt Bis-Tris Plus gels can be stored for 16 months at room temperature.Faster run times-separation of your proteins can be attained in 20-35 minutes.Wide ranges of molecular weight separation-selecting the right gel and running buffer helps to achieve the optimal separation of your proteins.Better protein integrity-optimized sample preparation process preserves your proteins.Features of the NuPAGE Bis-Tris and Bolt Bis-Tris Plus Gels include: The unique WedgeWell design of the Bolt Bis-Tris Plus gel sample well allows loading of up to 2x more sample volume than other precast gels and the Bis-Tris Plus chemistry enables Bolt gels to be run in as little as 20 minutes. Both NuPAGE Bis-Tris gels and Bolt Bis-Tris Plus gels are ideal for western blot analysis and applications such as sequencing and mass spectrometry where protein integrity is crucial and higher sensitivity is required. NuPAGE Bis-Tris gels are offered as mini and midi gels, and Bolt Bis-Tris Plus gels are offered as mini gels. preserving protein integrity becomes particularly important when separating low-abundance proteins. The Bis-Tris gel chemistry provides a neutral-pH environment that minimizes protein modifications, resulting in less protein degradation and sharper bands without typical gel 'smiling'. NuPAGE Bis-Tris and Bolt Bis-Tris Plus Gels are precast polyacrylamide gels designed to give optimal separation for a wide range of molecular weight proteins during gel electrophoresis under denaturing conditions. Membranes were imaged on an iBright Imaging System, with the same time and image adjustment settings for each blot. For fluorescence: multiplex detection, the membranes were probed overnight with a mixture of primary antibodies diluted in blocking solution: Rabbit-anti Vinculin (1:15,000), Rat anti-α Tubulin (1:5,000)), and Mouse-anti p23 (1:5,000) followed by an incubation with secondary antibodies diluted at 1:5,000 in 1X Blocker FL: Donkey anti-Rabbit Alexa Fluor Plus 680, Donkey anti-Rat Alexa Fluor 488, and Donkey anti-Mouse Alexa Fluor Plus 800 for 1 hour. Band loss and smearing can be seen at the higher loads for all targets on the Bio-Rad blot, while the blot for the NuPAGE Bis-Tris gel offers superior protein loading capacity above 24 µg and crisp, bright bands.įor immunodetection: membranes were blocked for 1 hour in 1X Blocker FL Fluorescent Blocking Buffer.

Both membranes were probed for three targets (vinculin, α-tubulin, and p23) using fluorescent immunodetection. Western blots using NuPAGE 4-12% Bis-Tris midi gels generate sharper bands at greater protein and RIPA lysis buffer loads than Bio-Rad 4–20% TGX midi gels.A NuPAGE 4–12% Bis-Tris midi gel, 12+2 well, was loaded with decreasing total protein amount of HEK293 lysate, subjected to electrophoresis in a SureLock Tandem Midi Gel Tank and transferred onto a 0.45 µm PVDF membrane using the SureLock Tandem Blot Module and the Bio-Rad 4–20% TGX midi gel, 12+2 well, was subjected to electrophoresis in a Criterion Midi Cell Tank and transferred onto a 0.45 µm PVDF membrane using the Criterion Blotter. *Not all percentages are available in every well type WedgeWell format: 10, 12, 15 17 well (load up to 2X sample volume per well) SureLock Tandem Midi Gel Tank, Invitrogen XCell4 SureLock Midi-Cell or Bio-Rad Criterion (with adapters only) Mini Gel Tank or XCell SureLock Mini-Cell NuPAGE MES SDS or NuPAGE MOPS SDS Running Bufferīolt MES SDS or Bolt MOPS SDS running buffer Working with dilute samples or need to load larger sample volumes, or when faster run times are neededĬoefficient of variation (CV) of only 2% for Rf values (migration) Need flexibility in well and gel format, or when you need a higher-throughput midi gel for a larger number of samples

Seeblue protein ladder Ctbp1 western blot