Kindlin Overexpression Lysate

Western Blot: Kindlin Overexpression Lysate (Adult Normal) [NBP2-06945] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Kindlin.

• Reactivity: Hu
• Applications: WB

Kindlin Overexpression Lysate Summary

• Description:

  Kindlin Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC217593
  
  Accession#: NM_017671
  
  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: FERMT1

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: 77.3 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 Kindlin Overexpression Lysate

• C20orf42fermitin family homolog 1
• chromosome 20 open reading frame 42
• fermitin family homolog 1 (Drosophila)
• fermitin family member 1
• FLJ20116
• KIND1DTGCU2
• kindlerin
• kindlin 1
• Kindlin syndrome protein
• Kindlin-1
• UNC112 related protein 1
• UNC112A
• Unc-112-related protein 1
• URP1FLJ23423

Background

Due to a concerted effort to identify biomarkers for lung and colon carcinomas by genome-wide transcriptional profiling, the identification and cloning of one such gene as well as two additional closely related genes was achieved. Due to the strong sequence homology to the C. elegans UNC-112 a novel gene gene was named URP1, for UNC-112 related protein. Another novel related gene, URP2 and the previously discovered MIG-2 gene was also identified. Transcriptional analysis shows that only URP1 is significantly differentially regulated, being over-expressed in 70% of the colon carcinomas and 60% of the lung carcinomas tested. Quantification of URP1 expression by qRT-PCR showed up-regulation of the gene by 60-fold in lung tumors and up to nearly 6-fold in colon tumors. Northern blot analysis of URP1 indicates that normal expression is restricted to neuromuscular tissues. In contrast, the expression of URP2 appears to be confined primarily to tissues of the immune system. URP1 has the potential to be one of the most important prognostic and diagnostic markers of colon or lung cancer.

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: FERMT1