a-Tubulin Antibody for Western Blotting

Biochemistry
Stanford University
Senior Research Scientist

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Company:

Cell Signaling Technology

Product Name:

α-Tubulin Antibody

Catalog Number:

2144

Rab proteins constitute the largest family of monomeric small GTPases that oscillate between active (GTP- bound) and inactive (GDP- bound) conformations. The spatio-temporal activation of Rabs is controlled by the GDP-GTP exchange factors (GEFs), mediating their activation, and the GTP hydrolysis activating proteins (GAPs), terminating their activity. Rab GTPases execute diverse tasks in membrane trafficking by driving cargo collection into nascent transport vesicles, linking vesicles to cytoskeletal elements, recruiting tethers, and finally docking vesicles to the target compartment, leading to membrane fusion. The goal of this project is to characterize a specific GEF for Golgi localized Rab6 GTPase, and to analyze its effect in Rab6-mediated processes at the Golgi complex. In the present experiment, alpha-tubulin antibody was used as a loading control for the gel. This antibody was recommended by a researcher from a neighboring lab.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Whole cell extracts of HeLa cells

Primary Incubation

1:5000 dilution in blocking buffer; 1 hr at room temp

Blocking Agent

5% Milk powder in 1xTBS-T(0.05%)

Secondary Incubation

Goat-anti Rabbit-IgG-HRP conjugated; 1:10000 dilution in blocking buffer; 1 hr at room temp

Tertiary Incubation

None

Detection

ECL regular; 2 sec exposure on a film

Results Summary

The loading of samples was uniform across three wells as judged by alpha-tubulin intensity.

Additional Notes

None

Related Categories

Image Gallery

Summary

The Good

Specific signal

The Bad

None

The Bottom Line

Alpha-tubulin antibody from Cell Signaling is a perfect antibody for Western Blotting.

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