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AKAP5 Overexpression Lysate

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Western Blot: AKAP5 Overexpression Lysate (Adult Normal) [NBL1-07429] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for AKAP5.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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AKAP5 Overexpression Lysate Summary

Description

AKAP5 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC221314

Accession#: NM_004857

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
AKAP5

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 (cat# 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
46.9 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 AKAP5 Overexpression Lysate

  • A kinase (PRKA) anchor protein 5
  • AKAP 79
  • AKAP7979kDa
  • A-kinase anchor protein 79 kDa
  • A-kinase anchoring protein 75/79
  • cAMP-dependent protein kinase regulatory subunit II high affinity bindingprotein
  • cAMP-dependent protein kinase regulatory subunit II high affinity-bindingprotein
  • H21

Background

The A-kinase anchor proteins (AKAPs) are a group of structurally diverse proteins, which have the common function of binding to the regulatory subunit of protein kinase A (PKA) and confining the holoenzyme to discrete locations within the cell. This gene encodes a member of the AKAP family. The encoded protein binds to the RII-beta regulatory subunit of PKA, and also to protein kinase C and the phosphatase calcineurin. It is predominantly expressed in cerebral cortex and may anchor the PKA protein at postsynaptic densities (PSD) and be involved in the regulation of postsynaptic events. It is also expressed in T lymphocytes and may function to inhibit interleukin-2 transcription by disrupting calcineurin-dependent dephosphorylation of NFAT. [provided by RefSeq]. Publication Note: This RefSeq record includes a subset of the publications that are available for this gene. Please see the Entrez Gene record to access additional publications.

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 AKAP5 Lysate (NBL1-07429) (0)

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Blogs on AKAP5.

Identifying tumoral and stromal transcriptomes that underlie tumor plasticity and stromal neuroinflammatory response in brain metastasis
By Jamshed Arslan, Pharm. D., PhD. Cancers in the brain often come from tumors elsewhere in the body. Several adaptive mechanisms influence brain metastasis, such as blood brain barrier leakage that can be induced by ...  Read full blog post.

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Bioinformatics

Gene Symbol AKAP5