Adrenomedullin/ADM Overexpression Lysate

Western Blot: Adrenomedullin/ADM Overexpression Lysate [NBL1-07345] - Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Adrenomedullin.

• Reactivity: Hu
• Applications: WB

Adrenomedullin/ADM Overexpression Lysate Summary

• Description:

  Adrenomedullin/ADM Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC200507
  
  Accession#: NM_001124
  
  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: ADM

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: 20.2 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 Adrenomedullin/ADM Overexpression Lysate

• ADM
• Adrenomedullin
• AMpreproadrenomedullin

Background

Adrenomedullin, a hypotensive peptide found in human pheochromocytoma, consists of 52 amino acids, has 1 intramolecular disulfide bond, and shows a slight homology with the calcitonin gene-related peptide. It may function as a hormone in circulation control because it is found in blood in a considerable concentration. The precursor, called preproadrenomedullin, is 185 amino acids long. By RNA-blot analysis, human adrenomedullin mRNA was found to be highly expressed in several tissues. Genomic ADM DNA consists of 4 exons and 3 introns, with the 5-prime flanking region containing TATA, CAAT, and GC boxes. There are also multiple binding sites for activator protein-2 and a cAMP-regulated enhancer element. [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 Adrenomedullin/ADM Lysate (NBL1-07345). (Showing 1 - 2 of 2 FAQ).

1. I have got a query regarding the following products: catalog # NBL1-07345 and catalog # H00000133-T04. Are they the same product/source?
   • NBL1-07345 and H00000133-T04 are both Adrenomedullin Overexpression Lysates but they are not the exact same products. H00000133-T04 is an Abnova product and NBL1-07345 is a Novus product. The main difference between these two is NBL1-07345 contains DDK/Myc tag at C-Terminus so you can use both Adrenomedullin and DDK antibody to detect the protein.
2. We are interested in the following references: NBL1-11754, NB820-59468, NBL1-14127,NBL1-07345. Have these references got any tag in their sequence for their purification, so the weight (KDa) increases?
   • NBL1-11754, NBL1-14127 and NBL1-07345 are all overexpression lysates; in each case, the protein contains a C-terminal Myc/DDK epitope tag (only 3 kDa total). The NB820-59468 is not an overexpression lysate; it is a whole normal brain (fetal) tissue lysate.

Bioinformatics

• Gene Symbol: ADM