TRPC3 Overexpression Lysate

Western Blot: TRPC3 Overexpression Lysate (Adult Normal) [NBL1-17331] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for TRPC3.

• Reactivity: Hu
• Applications: WB

TRPC3 Overexpression Lysate Summary

• Description:

  TRPC3 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC210794
  
  Accession#: NM_003305
  
  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: TRPC3

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: 97.2 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 TRPC3 Overexpression Lysate

• hTrp3
• hTrp-3
• short transient receptor potential channel 3
• transient receptor potential cation channel, subfamily C, member 3
• transient receptor potential channel 3
• Transient receptor protein 3
• TRP3
• TRP-3
• TRPC3

Background

The transient receptor potential canonical (TRPC) group of channels, including TPRC3, is a family of receptor-activated non-selective calcium permeant cation channels. In vitro TRPC proteins can form hetero- or homo-oligomeric channels. Endogenous TRPC1, TRPC3 and TRPC7 participate in forming heteromeric store-operated channels, while TRPC3 and TRPC7 can also participate in forming heteromeric receptor-operated channels. TRPC3 plays important roles in neuronal differentiation and immune cell maturation by mediating the cationic current in response to phospholipase C activation, Ca(2+) depletion, and diacylglycerol stimulation. TRPC3 channels are highly expressed in pontine neurons and mediate a nonselective cation current in response to the neurotrophin receptor, TrkB.

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 TRPC3 Lysate (NBL1-17331). (Showing 1 - 2 of 2 FAQ).

1. I would like to test some of your TRPC3 antibodies in my model system (Xenopus), but they have not been proven to work in this species. Do you offer any guarantee, or beta testing program. We would be happy to send you any images we produce if the antibody works (WB, IHC, ICC). We know that TRPC1-5 are expressed in our system (Xenopus spinal cord).
   • We do offer a testing program, the Innovators Reward Program. Our Innovator’s Reward program is designed to support your innovative research with minimal financial risk to you. Should you decide to use one of our products in an application or species for which it has not been tested all you need to do is submit a full review of the antibody with an image to qualify.
2. I am hoping you can help me concerning use of this lysate in Western Blots. How much of the lysate was loaded onto the gel used to generate the image seen on the data sheet?
   • For the image on our webpage for our TRPC3 overexpression lysate (NBL1-17331), we have loaded 10ug per lane. Please note that the blot was probed with an anti-DDK target, so your results may vary.

Bioinformatics

• Gene Symbol: TRPC3