MAP2 Overexpression Lysate

Western Blot: MAP2 Overexpression Lysate (Adult Normal) [NBP2-06122] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for MAP2.

• Reactivity: Hu
• Applications: WB

MAP2 Overexpression Lysate Summary

• Description:

  MAP2 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC221637
  
  Accession#: NM_031845
  
  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: MAP2

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: 49.5 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 MAP2 Overexpression Lysate

• DKFZp686I2148
• MAP2
• MAP-2
• MAP2A
• MAP2B
• MAP2C
• Microtubule Associated Protein 2
• microtubule-associated protein 2
• MTAP2

Background

Microtubule-associated protein 2 also known as MAP2 (theoretical molecular weight 280 kDa) belongs to the MAP2/Tau family which in addition to the two neuronal forms MAP2 and Tau also includes MAP4 (1). For MAP2, four different isoforms have been identified which are produced through alternative splicing including MAP2a, MAP2b, MAP2c and MAP2d (1). MAP2 isoforms contain multiple microtubule-binding domains located near the carboxy terminal, a variably sized amino-terminal projection domain, and a conserved protein kinase A binding domain. MAP2c and MAP2d isoforms have a shorter amino-terminal projection domain, lower molecular weight and are identifiable at ~70 kDa in SDS-PAGE analysis (2,3). MAP2's carboxy terminal domain supports interactions with microtubules, actin, and intermediate filaments (2).

MAP2 isoforms are developmentally regulated and differentially expressed in neurons and some glia. MAP2c is predominantly expressed in the developing brain while the other isoforms are expressed in the adult brain. The distribution of MAP2 isoforms also varies, with MAP2a and MAP2b predominantly localized to dendrites, while MAP2c is also found in axons. Lastly, the expression of MAP2d is not limited to neurons and may be found in glia, specifically oligodendrocytes (1, 2). MAP2 isoforms associate with microtubules and mediate their interaction with actin filaments thereby playing a critical role in organizing the microtubule-actin network. In neurons, MAP2 isoforms are implicated in different processes including neurite initiation, elongation and stabilization as well as axon and dendrite formation (2). Knockout of MAP expression in animal models results in a variety of functional and structural brain defects according to the isoform affected (e.g., reduced LTP and LTD, reduced myelination, absence of corpus collosum, motor system malfunction, abnormal hippocampal dendritic morphology, abnormal synaptic plasticity) (4).

References

1. Dehmelt, L., & Halpain, S. (2005). The MAP2/Tau family of microtubule-associated proteins. Genome Biology. https://doi.org/10.1186/gb-2004-6-1-204

2. Mohan, R., & John, A. (2015). Microtubule-associated proteins as direct crosslinkers of actin filaments and microtubules. IUBMB Life. https://doi.org/10.1002/iub.1384

3. Shafit-Zagardo, B., & Kalcheva, N. (1998). Making sense of the multiple MAP-2 transcripts and their role in the neuron. Molecular Neurobiology. https://doi.org/10.1007/BF02740642

4. Tortosa, E., Kapitein, L. C., & Hoogenraad, C. C. (2016). Microtubule organization and microtubule-associated proteins (MAPs). In Dendrites: Development and Disease. https://doi.org/10.1007/978-4-431-56050-0_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.

Bioinformatics

• Gene Symbol: MAP2