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Beta 2 Adaptin Overexpression Lysate

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Western Blot: Beta 2 Adaptin Overexpression Lysate (Adult Normal) [NBP2-11335] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Beta 2 Adaptin.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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Beta 2 Adaptin Overexpression Lysate Summary

Description

Beta 2 Adaptin Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC215791

Accession#: NM_001282

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
AP2B1

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
104.4 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 Beta 2 Adaptin Overexpression Lysate

  • Adapter-related protein complex 2 beta subunit
  • Adaptor protein complex AP-2 subunit beta
  • adaptor-related protein complex 2, beta 1 subunit
  • ADTB2adaptin, beta 2 (beta)
  • AP105B
  • AP2-BETA
  • beta-2-adaptin
  • beta-adaptin
  • CLAPB1AP-2 complex subunit beta
  • Clathrin assembly protein complex 2 beta large chain
  • clathrin-associated/assembly/adaptor protein, large, beta 1
  • DKFZp781K0743
  • Plasma membrane adaptor HA2/AP2 adaptin beta subunit

Background

Clathrin-mediated endocytosis is the pathway by which many receptors for nutrients and hormones are internalized to be recycled or down-regulated. During formation of clathrin coated membranes, clathrin co-assembles with heterotetrameric molecules known as assembly polypeptides (APs) or adaptors which form a layer of protein coat between the clathrin lattice and the membrane. There are two characterized adaptors AP1 and AP2. AP1 is associated with clathrin coated vesicles at the trans-Golgi network and AP2 is associated with the endocytic clathrin coated vesicles at the plasma membrane and has been shown to specifically interact with Shc and EGF receptor. AP2 is composed of four subunits, two separate 100 kDa gene products with similar domain structures (alpha and beta adaptin) and a 50 and 17 kDa subunit. There are two alpha-adaptin genes, alpha A and alpha C which have a tissue specific pattern of expression.

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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Bioinformatics

Gene Symbol AP2B1