Complement Component C5aR1 Overexpression Lysate

Western Blot: C5AR1 Overexpression Lysate (Adult Normal) [NBL1-08482] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for C5AR1.

• Reactivity: Hu
• Applications: WB

Complement Component C5aR1 Overexpression Lysate Summary

• Description:

  Complement Component C5aR1 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC203784
  
  Accession#: NM_001736
  
  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: C5AR1

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: 39.1 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 Complement Component C5aR1 Overexpression Lysate

• C5a anaphylatoxin chemotactic receptor
• C5A
• C5aR
• C5a-R
• C5AR1
• C5ARC5a anaphylatoxin receptor
• C5R1C5a ligand
• CD88 antigen
• CD88
• complement component 5 receptor 1 (C5a ligand)
• complement component 5 receptor 1
• complement component 5a receptor 1
• Complement Component C5a R1
• Complement Component C5aR1

Background

The human anaphylatoxin C5a is a 74 residue glycopeptide which is generated by proteolytic cleavage of the complement factor C5 in the course of complement activation. A variety of biological effects evoked by C5a can be demonstrated, rendering this molecule an important mediartor of inflammation, with granulocytes and macrophages as the main target cells. All cellular responses to C5a are specifically mediated by interactions with the membrane bound C5a receptor, a seven transmembrane GTP binding coupled receptor that belongs to the rhodopsin supergene family. C5a receptor 1 expression has been reported in myeloid blood cells, brain, liver, lung, spleen, heart, kidney, and intestinal tract. ESTs have been isolated from B cell/lung/testis, nerve, and placenta libraries.

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: C5AR1