Mutant-Specific Antibody

Mutation-specific antibodies are designed to recognize and bind to specific mutated forms of proteins. These mutations often occur in cancer cells, making mutation-specific antibodies valuable tools in both research and clinical settings. OriGene has created many mutant-specific antibodies highly specific to the targeted protein.

Gene Mutation Description SKU
BRAF V600E Anti-BRAF(V600E) Rabbit Monoclonal Antibody TA592461
KRAS G12D Anti-KRAS(G12D) Rabbit Monoclonal Antibody TA594368
IDH1 R132H Anti-IDH1(R132H) Rat Monoclonal Antibody TA190113

Identification of Mutant Antibody with CytoSections™

What are CytoSections?

OriGene has developed a tool for antibody validation called CytoSections. CytoSections are FFPE sections of transiently transfected cDNA specific gene targets overexpressed in mammalian cells. They are a cost-effective, easily renewable, and unlimited source of controls for validating mutant-specific antibodies. Below, we describe how OriGene used CytoSections to validate an antibody specific to the key IDH1(R132H) mutation for use in IHC assays.

CytoSections either over-expressing IDH1 or mutant IDH1( R132H) and untransfected CytoSections were used to screen two different mutant-specific antibodies against IDH1(R132H). The results are shown below

CytoSection Antibody Table
Use of the DDK antibody (TA592569), resulted in positive (brown) staining, in WT (IDH1) and mutant (IDH1(R132H) CytoSections (1A and 1B) respectively). This demonstrates that the CytoSections are expressing either the DDK-tagged IDH1 WT construct, or the mutant construct, as expected. There is no positive staining in the untransfected negative control (1C). WT IDH1 antibody (TA800406) detected both WT (2A) and mutant IDH1(R132H) (2B) while the untransfected control (2C) is negative.

Development and Specificity

It is difficult to develop mutant-specific antibodies. The key factors to keep in mind are:

Targeted Mutations: Mutation-specific antibodies are engineered to target proteins that have undergone specific point mutations, insertions, deletions, or other alterations. These mutations can alter the protein's structure, creating unique epitopes (binding sites) that differ from the wild-type (normal) protein.

Precision: These antibodies are highly specific, meaning they can distinguish between the normal and mutated forms of a protein. This specificity allows for precise targeting of cancer or other diseased cells while sparing normal, healthy cells.

Mutant Antibodies from OriGene

Mutant Antibodies from OriGene Mutant Antibodies from OriGene Mutant Antibodies from OriGene
Figure A, Western blot analysis of overexpressed lysate from HEK293T
cells transfected with empty plasmid (PS100001, lane NC), BRAF V600E mutant plasmid (RC400155, lane V600E) and BRAF wild type plasmid (RC211013, lane WT) using anti-BRAF V600E antibody TA592461 (1:500). Figure B, Western blot analysis of the same samples as figure A with anti-DDK antibody (TA180144, 1:1000)
HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane "293T")
or pCMV6-ENTRY IDH1 (wild type RC210582, middle lane, ) or pCMV6-ENTRY IDH1 mutated ( R132H mutation RC400096, right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (10 ug per lane) were separated by SDS-PAGE and immunoblotted with TA190113 (1:500) and then goat anti-rat IgG-HRP (1:2000).
Figure A, Western blot analysis of overexpressed lysates(15ug per lane)
from HEK293T cells transfected with empty plasmid (PS100001, NC) , human KRASva plasmid (RC222697, KRASva), human KRASvb plasmid (RC201958, KRASvb), KRAS(G12C) plasmid (RC400107, KRAS(G12C)), KRAS(G12V) plasmid (RC400109, KRAS(G12V)), KRAS(G12D) plasmid (RC400104, KRAS(G12D))using anti-KRAS mutant (G12D) antibody TA594368(1:100). Figure B, Western blot analysis of the same samples as figure A with anti-DDK antibody (TA180144, 1:1000)

BRAF (V600E)

IDH1 (R132H)

KRAS (G12D)

Antibody Resources