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GAP-43 Overexpression Lysate

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Western Blot: GAP43 Overexpression Lysate (Adult Normal) [NBP2-04214] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for GAP43.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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GAP-43 Overexpression Lysate Summary

Description

GAP-43 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC203175

Accession#: NM_002045

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
GAP43

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
24.6 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 GAP-43 Overexpression Lysate

  • Axonal membrane protein GAP-43
  • B-50
  • Basp2
  • calmodulin-binding protein P-57
  • GAP43
  • GAP-43
  • growth associated protein 43
  • Growth-associated protein 43
  • nerve growth-related peptide GAP43
  • Neural phosphoprotein B-50
  • Neuromodulin
  • neuron growth-associated protein 43
  • PP46
  • protein F1

Background

GAP43 (Growth associated protein 43) is a crucial component for regenerative response in the nervous system. GAP-43 is normally produced by neurons during developmental growth and axonal regeneration, but it is also expressed in specific regions of the normal adult nervous system. The protein is present at high levels in neuronal growth cones during development and axonal regeneration.

Heterozygous GAP3 knockout mice with GAP-43 levels reduced by one-half display significant memory impairments in cued conditioning, or on tests of nociceptive or auditory perception. Differentiating neurons that do not express GAP-43 show mislocalization of the centrosome and mitotic spindles, particularly in neurogenic cell divisions. Therefore, the neuronal precursor pool in the cerebellum fails to grow resulting in a cerebellum that is significantly smaller than normal. Total GAP-43 deletion is lethal within days of birth, demonstrating that GAP-43 is critical for the proper development of the mammalian CNS.

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 GAP43