SP-D Overexpression Lysate

Western Blot: Surfactant protein D Overexpression Lysate (Adult Normal) [NBL1-15897] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Surfactant protein D.

• Reactivity: Hu
• Applications: WB

SP-D Overexpression Lysate Summary

• Description:

  SP-D Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC204746
  
  Accession#: NM_003019
  
  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: SFTPD

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: 35.4 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 SP-D Overexpression Lysate

• COLEC7collectin-7
• Collectin 7
• Collectin-7
• Lung surfactant protein D
• PSPD
• PSP-D
• SFTP4
• SFTP4pulmonary surfactant-associated protein D
• SFTPD
• SPD
• SP-D
• SP-Dpulmonary surfactant apoprotein
• surfactant protein D
• surfactant, pulmonary-associated protein D
• surfactant-associated protein, pulmonary 4

Background

Surfactant protein D (SP-D) is synthesized and secreted by lung epithelial cells. It belongs to group III of the family of C-type lectins and members of this group has overall structure consisting of multiple globular head regions linked by triple-helical, collagen-like, strands. This group also includes SP-A and the serum proteins mannan-binding protein, conglutinin and collectin-43, all of which have been shown to bind to the C1q receptor found on a wide variety of cells. Both SP-D and SP-A have been shown to enhance oxygen radical production by alveolar macrophages. The serum concentration is 88 ng/ml in healthy individuals (2).

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: SFTPD