IL-32 Overexpression Lysate

Western Blot: IL32 Overexpression Lysate (Adult Normal) [NBP2-08698] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for IL32.

• Reactivity: Hu
• Applications: WB

IL-32 Overexpression Lysate Summary

• Description:

  IL-32 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC215952
  
  Accession#: NM_001012631
  
  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: IL32

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: 21.6 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 IL-32 Overexpression Lysate

• IL32
• IL-32
• IL-32alpha
• IL-32beta
• interleukin 32
• interleukin-32 theta
• interleukin-32
• natural killer cell transcript 4
• Natural killer cells protein 4
• NK4
• NK4IL-32delta
• TAIF
• TAIFa
• TAIFb
• TAIFc
• TAIFd
• TAIFIL-32gamma
• Tumor necrosis factor alpha-inducing factor

Background

IL32 alfa is the shortest of the 4 isotypes of IL32 known, it is 134 amino acids ( kDa) protein, the IL32B (beta) is 188, IL32C is 168 and IL32D is 179 amino acids long proteins. Each subtype has an N-terminal segment and four kringle domains (NK4), that interact with several other proteins including the hepatic growth factor via c-met a tyrosine receptor kinase (5). IL-32 synergized with the intracellular nuclear oligomerization domain receptors (NOD1- and NOD2-specific muropeptides of peptidoglycans for the release of IL-1beta and IL-6. In contrast, IL-32 did not influence the cytokine production induced via TLRs (6). The anti-IL32 selective antibodies were made against an epitope that lies near the C-terminal end of the protein. The antibodies to IL32 are affinity purified on immobilized affinity based chromatography and characterized for applications in ELISA and Western blotting. The antiIL32 antibodies recognize a single band of IL32 in PC-IL32 samples. The IL32 antibodies do not cross react with other pro-inflammatory or anti-inflammatory interleukins, has also produced antibodies to other interleukins, interleukin receptors, cytokines and their receptors.

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: IL32