Alpha-mannosidase II Overexpression Lysate

Western Blot: Alpha-mannosidase II Overexpression Lysate (Adult Normal) [NBL1-12834] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Alpha-mannosidase II.

• Reactivity: Hu
• Applications: WB

Alpha-mannosidase II Overexpression Lysate Summary

• Description:

  Alpha-mannosidase II Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC220186
  
  Accession#: NM_002372
  
  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: MAN2A1

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: 131 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 Alpha-mannosidase II Overexpression Lysate

• alpha-mannosidase 2
• AMan II
• EC 3.2.1
• EC 3.2.1.114
• Golgi alpha-mannosidase II
• golgi integral membrane protein 7
• GOLIM7
• Man II
• MANA2MANII
• Mannosidase alpha class 2A member 1
• mannosidase, alpha type II
• mannosidase, alpha, class 2A, member 1
• Mannosyl-oligosaccharide 1,3-1,6-alpha-mannosidase

Background

Alpha-mannosidase II encodes a protein which is a member of family 38 of the glycosyl hydrolases. The protein is located in the Golgi and catalyzes the final hydrolytic step in the asparagine-linked oligosaccharide (N-glycan) maturation pathway. Mutations in the mouse homolog of this gene have been shown to cause a systemic autoimmune disease similar to human systemic lupus erythematosus. [provided by RefSeq]

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.

FAQs for Alpha-mannosidase II Lysate (NBL1-12834). (Showing 1 - 1 of 1 FAQ).

1. How much lysate is used in WB example? How much is recommended for positive control?
   • The Western blot image you see shows 5ug of protein. 5-10ug of lysate may be used depending on the sensitivity of the antibody.

Bioinformatics

• Gene Symbol: MAN2A1