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LAT Overexpression Lysate

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Western Blot: LAT Overexpression Lysate (Adult Normal) [NBP2-09156] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for LAT.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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LAT Overexpression Lysate Summary

Description

LAT Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC204180

Accession#: NM_001014987

Protein Tag: C-MYC/DDK

You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.
Gene
LAT

Applications/Dilutions

Dilutions
  • Western Blot
Application Notes
This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.

Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.
Theoretical MW
24.8 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

Storage
Store at -80C. Avoid freeze-thaw cycles.
Buffer
RIPA buffer

Lysate Details for Array

Type
Overexpression

Notes

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

Alternate Names for LAT Overexpression Lysate

  • 36 kDa phospho-tyrosine adapter protein
  • LAT
  • LAT1,36 kDa phospho-tyrosine adaptor protein
  • linker for activation of T cells
  • linker for activation of T-cells family member 1
  • p36-38
  • pp36

Background

LAT (linker for activation of T-cells) is involved in the T-cell antigen receptor (TCR) signal transduction pathway, and may play an important role downstream in activating protein tyrosine kinases (1). LAT is phosphorylated by ZAP-70/Syk protein tyrosine kinases leading to recruitment of multiple signaling molecules (2). It has been found that Tyr191 and Tyr171 are required for T-cell activation and Tyr132 is required for the activation of PLCgamma1 and Ras signaling pathways, respectively. Tyr226 and Tyr191 are required for Vav binding, whereas Tyr171 and Tyr132 are necessary for association and activation of phosphoinositide 3-kinase activity and PLCgamma1 (3).

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.

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Publications for LAT Lysate (NBP2-09156) (0)

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Product General Protocols

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Bioinformatics

Gene Symbol LAT