Difference between revisions of "Part:BBa K3573001:Experience"

(→‎Applications of BBa_K3573001)
(→‎Applications of BBa_K3573001)
Line 6: Line 6:
 
===Applications of BBa_K3573001===
 
===Applications of BBa_K3573001===
  
−
Our team has cloned scFv(RAS) into an expression vector and tested the protein expression in 4 different conditions (2 temperatures X 2 IPTG concentrations)
+
Our team has amplified scFv(RAS) using PCR and cloned the gene into an expression vector.
  
−
After expression, we used Ni-NTA column for protein purification using the His6-tag located at the N-terminus of CPP-scFv(RAS). We have confirmed the identity of the protein using Western blot analysis using anti-His6 antibody.
+
[[File:T--Korea_HS--scFvPCR.jpg|500px]]
  
−
[[File:T--Korea_HS--1.png|500px]]
+
 
 +
We have tested protein expression in 4 different conditions (2 temperatures X 2 IPTG concentrations). After expression, Ni-NTA column was used for protein purification using the His6-tag located at the N-terminus of CPP-scFv(RAS).
 +
 
 +
[[File:T--Korea_HS--scFvNiNTA.jpg|500px]]
 +
 
 +
 
 +
 
 +
We have confirmed the identity of the protein using Western blot analysis using anti-His6 antibody.
 +
 
 +
[[File:T--Korea_HS--scFvWestern.jpg|500px]]
 +
 
 +
 
 +
 
 +
HRas(G12V) that is described part number BBa_K3573002 was used for binding assay with scFv(RAS) using size-exclusion chromatography.
 +
 
 +
[[File:T--Korea_HS--scFvRASBinding.jpg|500px]]
  
 
===User Reviews===
 
===User Reviews===

Revision as of 23:55, 25 October 2020


This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K3573001

Our team has amplified scFv(RAS) using PCR and cloned the gene into an expression vector.

T--Korea HS--scFvPCR.jpg


We have tested protein expression in 4 different conditions (2 temperatures X 2 IPTG concentrations). After expression, Ni-NTA column was used for protein purification using the His6-tag located at the N-terminus of CPP-scFv(RAS).

T--Korea HS--scFvNiNTA.jpg


We have confirmed the identity of the protein using Western blot analysis using anti-His6 antibody.

T--Korea HS--scFvWestern.jpg


HRas(G12V) that is described part number BBa_K3573002 was used for binding assay with scFv(RAS) using size-exclusion chromatography.

T--Korea HS--scFvRASBinding.jpg

User Reviews

UNIQd313d9a71d1ccae7-partinfo-00000000-QINU UNIQd313d9a71d1ccae7-partinfo-00000001-QINU