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ASPP1 Overexpression Lysate

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Western Blot: ASPP1 Overexpression Lysate (Adult Normal) [NBP2-10288] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for ASPP1.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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ASPP1 Overexpression Lysate Summary

Description

ASPP1 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC221172

Accession#: NM_015316

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
PPP1R13B

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
119.4 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 ASPP1 Overexpression Lysate

  • apoptosis-stimulating protein of p53, 1
  • ASPP1Protein phosphatase 1 regulatory subunit 13B
  • KIAA0771apoptosis-stimulating of p53 protein 1
  • p53BP2-like
  • p85
  • protein phosphatase 1, regulatory (inhibitor) subunit 13B

Background

The ASPP family members ASPP1, ASPP2, act as potent activators of p53. A third member of the family, iASPP, has an inhibitory effect on apoptosis. The three members of ASPP family all bind p53 via their C-terminus, which contains a proline-rich region, four ankyrin repeats and an SH3 domain. Expression of ASPP1 and ASPP2 is frequently downregulated in human breast tumors expressing wild-type p53. ASPP1 may also play a role in vascular development.

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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Video Protocols

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Bioinformatics

Gene Symbol PPP1R13B