S100A9 Overexpression Lysate

Western Blot: S100A9 Overexpression Lysate (Adult Normal) [NBL1-15660] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for S100A9.

• Reactivity: Hu
• Applications: WB

S100A9 Overexpression Lysate Summary

• Description:

  S100A9 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC208250
  
  Accession#: NM_002965
  
  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: S100A9

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: 13.1 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 S100A9 Overexpression Lysate

• CAGBMigration inhibitory factor-related protein 14
• Calgranulin B
• calgranulin-B
• Calprotectin L1H subunit
• CFAGMRP-14,60B8AG
• CGLB
• L1AG
• LIAG
• MAC387
• MIF
• MRP-14
• MRP14Leukocyte L1 complex heavy chain
• NIF
• P14
• protein S100-A9
• S100 calcium binding protein A9 (calgranulin B)
• S100 calcium binding protein A9
• S100 calcium-binding protein A9 (calgranulin B)
• S100 calcium-binding protein A9
• S100A9

Background

The protein encoded by this gene is a member of the S100 family of proteins containing 2 EF-hand calcium-binding motifs. S100 proteins are localized in the cytoplasm and/or nucleus of a wide range of cells, and involved in the regulation of a number of cellular processes such as cell cycle progression and differentiation. S100 genes include at least 13 members which are located as a cluster on chromosome 1q21. This protein may function in the inhibition of casein kinase and altered expression of this protein is associated with the disease cystic fibrosis. [provided by RefSeq]. Publication Note: This RefSeq record includes a subset of the publications that are available for this gene. Please see the Entrez Gene record to access additional publications.

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 S100A9 Lysate (NBL1-15660). (Showing 1 - 2 of 2 FAQ).

1. Have any of your S100A9 antibodies been tested for use in neutralization, or blocking assays, on human samples?
   • Unfortunately at this time we have not tested, or received customer feedback on our S100A9 antibodies for use in blocking or neutralizing assays. If you were planning on testing any of our S100A9 antibodies for neutralizing or blocking capabilities, we would recommend our Innovator’s Reward Program. Under the terms of this program we would be happy to provide a credit for a free vial in exchange for new data on this previously untested or reported application. Please submit this data in the form of an online review. Additional <a href="http://www.novusbio.com/support/innovators-reward.html" target="_self">Innovators Reward Program</a> information can be found using this link.
2. We would like to stain paraffin embedded human intestinal tissue for Calprotectin. One question will be which cells express the majority of Calprotectin - invading neutrophils vs epithelium for example. The antibody list you provide is quite extensive. Which antibody would you recommend for our purpose? Which tissue would you recommend as positive control.
   • Nine of our antibodies to Calprotectin have been validated for IHC-P with human tissue, and you can see all of these products. A number of our IHC images were generated using human spleen samples, and as such this may be a good choice of a positive control tissue. We do sell a human spleen slide product, which is suitable for IHC-P. Unfortunately I do not have an in-depth knowledge of Calprotectin, however the image shown for our antibody with catalogue number NBP1-02826 demonstrates clear staining of neutrophils. The following paper also suggests that the protein is abundant in neutrophils: PMID 11435495.

Bioinformatics

• Gene Symbol: S100A9