Rab5a Knockout HeLa Cell Lysate

Product Discontinued

Rab5a Knockout HeLa Cell Lysate Summary

• Preparation Method: Knockout achieved by using CRISPR/Cas9,Insertion of the selection cassette in exon 2
• Gene: RAB5A

Applications/Dilutions

• Dilutions:
  • Western Blot
• Application Notes: You will receive 1 vial (100ug) of knockout cell lysate and 1 vial (100ug) of Parental cell lysate. Lysate can be diluted with 1X SDS sample buffer and will be stable at -20 degrees C for 12 months. Minimize freeze-thaw cycles.

Packaging, Storage & Formulations

• Storage: Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles.
• Buffer: 0.1 mg cell homogenate lyophilized in RIPA buffer made with double-knockout cell lines.
• Concentration: LYOPH
• Reconstitution Instructions: To use as WB negative control, spin down briefly and resuspend in 100 uL 1xSDS sample buffer (2% SDS, 60 mM Tris-HCl pH 6.8, 10% Glycerol, 0.02% Bromophenol blue, 60 mM beta-mercaptoethanol). Boil the lysate for 3 - 5 minutes before loading it onto gel.

Lysate Details for Array

• Type: Knockout HeLa Cell

Notes

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Validation of antibody specificity is critical and verification of antibody performance against knockout samples is one way to guarantee that an antibody recognizes a specific target. Novus' KO cell lysate can be used as a negative control for western blots and to confirm the specificity of antibodies.

Alternate Names for Rab5a Knockout HeLa Cell Lysate

• RAB5A, member RAS oncogene family
• RAB5RAS-associated protein RAB5A
• ras-related protein Rab-5A

Background

Rab5 is a 24 kDa GTP-binding protein that regulates the fusion of plasma membrane -derived clathrin-coated vesicles with early endosomes , and homotypic fusion among early endosomes1. Localized to the cytoplasmic side of the plasma membrane, clathrin -coated vesicles and early endosomes, Rab5 appears to regulate vesicle fusion through a cycle of GDP/GTP e xchange and GTP hydrolysis. The different guanine nuc leotide binding states of Rab5 may affect its ability to associate or dissociate with membranes during endocytotic membrane traffic. The GTP-bound, or active, form of Rab5 associates with membrane s and regulates vesicle docking and fusion. Studies using a Rab5 mutant that hydrolyzed xanthosine 5 -triphosphate (XTP) indicated that nucleotide hydrolysis occurs even in the absence of membrane fusion. GTP hydrolysis by Rab5 may determine the frequency of membrane docking and fusion events.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Bioinformatics

• Gene Symbol: RAB5A