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krasg12v  (Addgene inc)


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    Addgene inc krasg12v
    Krasg12v, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 30 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/krasg12v/pBABE-Puro-KRas*G12V+(Plasmid+%2346746)/pm40935852-55-13-22
    Average 93 stars, based on 30 article reviews
    krasg12v - by Bioz Stars, 2026-11
    93/100 stars

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    Stable Transfection:

    Article Title: Mutant KRAS triggers functional reprogramming of tumor-associated macrophages in colorectal cancer
    Article Snippet: Transfection of HIF-1α-specific siRNAs in target cells was performed using a heterogeneous mixture of siRNAs (Sigma). .. To establish SW48 cells stably expressing the KRASG12V mutation, SW48 cells were stably transfected with plasmids encoding either an empty vector (pBabe-puro, Addgene) or KRASG12V (pBabe-puro-KRAS12V, Addgene) using the Amaxa system (Lonza). .. Cells were selected with puromycin (1μg/ml, Merck) for 10 days.

    Article Title: Loss of MLL Induces Epigenetic Dysregulation of Rasgrf1 to Attenuate Kras-Driven Lung Tumorigenesis
    Article Snippet: .. Establishment of KrasG12V stably overexpressing cell line Lenti-mCherry-G12V-Kras plasmid was purchased from Addgene and used to generate stable cell lines. ..

    Expressing:

    Article Title: Mutant KRAS triggers functional reprogramming of tumor-associated macrophages in colorectal cancer
    Article Snippet: Transfection of HIF-1α-specific siRNAs in target cells was performed using a heterogeneous mixture of siRNAs (Sigma). .. To establish SW48 cells stably expressing the KRASG12V mutation, SW48 cells were stably transfected with plasmids encoding either an empty vector (pBabe-puro, Addgene) or KRASG12V (pBabe-puro-KRAS12V, Addgene) using the Amaxa system (Lonza). .. Cells were selected with puromycin (1μg/ml, Merck) for 10 days.

    Article Title: Single-cell analyses reveal YAP/TAZ as regulators of stemness and cell plasticity in Glioblastoma
    Article Snippet: For constitutive expression of RTKs, cDNAs of HER2CA (from FUW-tetO-HER2-CA), PDGFRαCA (from pcDNA5FRT-EF-Pdgfrα-CA-EGFPN, Addgene#66789) and EGFRCA (from pBabe EGFR-L858R/T790M, Addgene#32073) were subcloned in CSII-CMV-MCS-IRES-puro empty vector backbone (obtained by substituting the blasticidin (bsd) resistance of CSII-CMV-MCS-IRES-bsd (a gift of H. Miyoshi) with the puromycin resistance). .. For constitutive lentiviral expression of mutant KRas, the pTomo KRasG12V was generated by subcloning the KRasG12V from the pBabe KRasG12V (Addgene#46746) to the pTomo-MCS Empty vector, obtained by substituting the loxP-RFP-loxP cassette from the pTomo vector (Addgene #26291) with a synthetic multiple cloning sites (MCS). .. For constitutive lentiviral expression of mutant KRas, the pTomo KRasG12V was generated by subcloning the KRasG12V from the pBabe KRasG12V (Addgene#46746) to the pTomo-MCS Empty vector, obtained by substituting the loxP-RFP-loxP cassette from the pTomo vector (Addgene #26291) with a synthetic multiple cloning sites (MCS).

    Article Title: Salmonella -based platform for efficient delivery of functional binding proteins to the cytosol
    Article Snippet: List of plasmids used in this study Plasmid Description pCASP-SptP1673G124-HilA triggers secretion of SptP167 fused to a-GFP DARPin from Salmonella upon induction with arabinose pCASP-SptP1203G124-HilA triggers secretion of SptP120 fused to a-GFP DARPin from Salmonella upon induction with arabinose pCASP-3G124-HilA expression of a-GFP DARPin from Salmonella is disrupted pCASP-SptP1203m160-HilA triggers secretion of SptP120 fused to a-mCherry DARPin from Salmonella upon induction with arabinose pCASP-SptP120E3_5-HilA triggers secretion of SptP120 fused to E3_5 DARPin from Salmonella upon induction with arabinose pCASP-SptP120NS1-HilA triggers secretion of SptP120 fused to NS1 monobody from Salmonella upon induction with arabinose pCASP-SptP120K27-HilA triggers secretion of SptP120 fused to K27 DARPin from Salmonella upon induction with arabinose pCASP-SptP120K55-HilA triggers secretion of SptP120 fused to K55 DARPin from Salmonella upon induction with arabinose pCASP-SptP120-TDHilA triggers secretion of SptP120 fused to HA4-7c12 Tandem Monobody from Salmonella upon induction with arabinose pCASP-SptP92-TDHilA triggers secretion of SptP92 fused to HA4-7c12 Tandem Monobody from Salmonella upon induction with arabinose pCASP-SptP35-TDHilA triggers secretion of SptP35 fused to HA4-7c12 Tandem Monobody from Salmonella upon induction with arabinose pCASP-HilA empty vector triggers HilA overexpression upon induction with arabinose without any synthetic protein expressed from Salmonella pCMV R8.74 lentiviral packaging plasmid – gift from Didier Trono (Addgene plasmid # 22036) pMD2.G vesicular stomatitis G envelope expressing plasmid – gift from Dider Trono (Addgene plasmid # 12259) Lenti-AcGFP-Sec61IRES-Blast vector plasmid for integration and constitutive expression of AcGFP fused to Sec61 in mammalian cells – gift from D. Gerlich (IMBA, Vienna). .. Lenti-mCherry-WTKRAS-IRES-Blast vector plasmid for integration and constitutive expression of mCherry fused to wild-type KRAS in mammalian cells Lenti-mCherry-G12VKRAS-IRES-Blast vector plasmid for integration and constitutive expression of mCherry fused to KRASG12V in mammalian cells All plasmid maps and sequences can be found on Addgene.org Supplementary Table 4. .. List of cell lines used in this study Cell line Description HCT116 Human colorectal carcinoma derived cell line bearing a KRASG13D activating mutation – obtained from ATCC HCT116 G12V KRAS-mCherry HCT116 cells engineered to stably express mutated G12V KRAS fused to mCherry – created in this study by lentiviral transduction HCT116 Sec61-GFP HCT116 cells engineered to stably express Sec61 fused to AcGFP – created in this study by lentiviral transduction HeLa HeLa Kyoto – human cervix adenocarcinoma derived cell line – gift from the lab of D. Gerlich (IMBA, Vienna) HeLa Sec61-GFP HeLa Kyoto cells engineered to stably express Sec61 fused to AcGFP – gift from the lab of D. Gerlich (unpublished) HeLa H2B-GFP HeLa Kyoto cells engineered to stably express H2B fused to EGFP – gift from the lab of D. Gerlich ref. 6 HeLa alpha-Tubulin-mCherry HeLa Kyoto cells engineered to stably express alpha-Tubulin fused to mCherry – gift from the lab of D. Gerlich ref. 6 HeLa Sec61-GFP H2B-mCherry HeLa Kyoto cells engineered to stably express Sec61 fused to AcGFP and H2B fused to mCherry – created in this study by lentiviral transduction of HeLa Kyoto cells that also stably express H2B-mCherry.

    Mutagenesis:

    Article Title: Mutant KRAS triggers functional reprogramming of tumor-associated macrophages in colorectal cancer
    Article Snippet: Transfection of HIF-1α-specific siRNAs in target cells was performed using a heterogeneous mixture of siRNAs (Sigma). .. To establish SW48 cells stably expressing the KRASG12V mutation, SW48 cells were stably transfected with plasmids encoding either an empty vector (pBabe-puro, Addgene) or KRASG12V (pBabe-puro-KRAS12V, Addgene) using the Amaxa system (Lonza). .. Cells were selected with puromycin (1μg/ml, Merck) for 10 days.

    Article Title: Single-cell analyses reveal YAP/TAZ as regulators of stemness and cell plasticity in Glioblastoma
    Article Snippet: For constitutive expression of RTKs, cDNAs of HER2CA (from FUW-tetO-HER2-CA), PDGFRαCA (from pcDNA5FRT-EF-Pdgfrα-CA-EGFPN, Addgene#66789) and EGFRCA (from pBabe EGFR-L858R/T790M, Addgene#32073) were subcloned in CSII-CMV-MCS-IRES-puro empty vector backbone (obtained by substituting the blasticidin (bsd) resistance of CSII-CMV-MCS-IRES-bsd (a gift of H. Miyoshi) with the puromycin resistance). .. For constitutive lentiviral expression of mutant KRas, the pTomo KRasG12V was generated by subcloning the KRasG12V from the pBabe KRasG12V (Addgene#46746) to the pTomo-MCS Empty vector, obtained by substituting the loxP-RFP-loxP cassette from the pTomo vector (Addgene #26291) with a synthetic multiple cloning sites (MCS). .. For constitutive lentiviral expression of mutant KRas, the pTomo KRasG12V was generated by subcloning the KRasG12V from the pBabe KRasG12V (Addgene#46746) to the pTomo-MCS Empty vector, obtained by substituting the loxP-RFP-loxP cassette from the pTomo vector (Addgene #26291) with a synthetic multiple cloning sites (MCS).

    Transfection:

    Article Title: Mutant KRAS triggers functional reprogramming of tumor-associated macrophages in colorectal cancer
    Article Snippet: Transfection of HIF-1α-specific siRNAs in target cells was performed using a heterogeneous mixture of siRNAs (Sigma). .. To establish SW48 cells stably expressing the KRASG12V mutation, SW48 cells were stably transfected with plasmids encoding either an empty vector (pBabe-puro, Addgene) or KRASG12V (pBabe-puro-KRAS12V, Addgene) using the Amaxa system (Lonza). .. Cells were selected with puromycin (1μg/ml, Merck) for 10 days.

    Enzyme-linked Immunosorbent Assay:

    Article Title: Targeting TGF-β-Smad2/3-JNK1-mediated SIRT1 activity overcomes the chemoresistance of KRAS mutation lung cancer.
    Article Snippet: .. Plasmids, antibodies, chemicals and ELISA assay kits KRASWT (#75282), KRASG12C (#58901), KRASG12D (#58902), KRASG12V (#46746) and JNK1-GFP (#86830) plasmids were purchased from Addgene. .. Small interfering RNAs (siRNAs), including siKRAS (sc-35731 and sc-43874), siSIRT1 (sc-40986) and siSmad2/3 (sc-37238) and antibodies including SIRT1 (sc-15454), KRAS (F234, sc-30), PARP (sc-8007) and cleaved PARP (sc56196) were purchased from Santa Cruz Biotechnology.

    Article Title: Targeting TGF-β-Smad2/3-JNK1-mediated SIRT1 activity overcomes the chemoresistance of KRAS mutation lung cancer
    Article Snippet: .. Plasmids, antibodies, chemicals, and ELISA assay kits KRAS WT (#75282), KRASG12C (#58901), KRASG12D (#58902), KRASG12V (#46746), and JNK1-GFP (#86830) plasmids were purchased from Addgene (Watertown, MA, USA). .. Small interfering RNAs (siRNAs), including siKRAS (sc-35731 and sc-43874), siSIRT1 (sc-40986), and siSmad2/3 (sc-37238), and antibody including SIRT1 (sc-15454), KRAS (F234, sc-30), PARP (sc-8007), and cleaved PARP (sc56196) were purchased from Santa Cruz Biotechnology Inc. (Dallas, TX, USA).

    Over Expression:

    Article Title: Genetically Defined Syngeneic Mouse Models of Ovarian Cancer as Tools for the Discovery of Combination Immunotherapy
    Article Snippet: Mouse Akt2 (Addgene plasmid #64832) was cloned into pLV-EF1aIRES-Blast (Addgene plasmid #85133). .. For Myc overexpression, we used MSCV-Myc-PGKPuro-IRES-GFP (Addgene plasmid #75124), and for KrasG12V, we used pUG2K (Addgene plasmid #35493). .. The lentiviral vector used to overexpress Brd4-short isoform, Smarca4 and Follistatin (Fst) in our study, pLV[Exp]-EF1A>mBrd4[NM_198094.2](ns):P2A:EGF and pLV[Exp]EF1A>mSmarca4 [NM_001357764.1] (ns):P2A:EGFP and pLV[Exp]-mCherryEF1A>mFst[NM_001301373.1] (VB200220-1171ukt) overexpression plasmids were used that were constructed by VectorBuilder.

    Plasmid Preparation:

    Article Title: Genetically Defined Syngeneic Mouse Models of Ovarian Cancer as Tools for the Discovery of Combination Immunotherapy
    Article Snippet: Mouse Akt2 (Addgene plasmid #64832) was cloned into pLV-EF1aIRES-Blast (Addgene plasmid #85133). .. For Myc overexpression, we used MSCV-Myc-PGKPuro-IRES-GFP (Addgene plasmid #75124), and for KrasG12V, we used pUG2K (Addgene plasmid #35493). .. The lentiviral vector used to overexpress Brd4-short isoform, Smarca4 and Follistatin (Fst) in our study, pLV[Exp]-EF1A>mBrd4[NM_198094.2](ns):P2A:EGF and pLV[Exp]EF1A>mSmarca4 [NM_001357764.1] (ns):P2A:EGFP and pLV[Exp]-mCherryEF1A>mFst[NM_001301373.1] (VB200220-1171ukt) overexpression plasmids were used that were constructed by VectorBuilder.

    Article Title: Salmonella -based platform for efficient delivery of functional binding proteins to the cytosol
    Article Snippet: List of plasmids used in this study Plasmid Description pCASP-SptP1673G124-HilA triggers secretion of SptP167 fused to a-GFP DARPin from Salmonella upon induction with arabinose pCASP-SptP1203G124-HilA triggers secretion of SptP120 fused to a-GFP DARPin from Salmonella upon induction with arabinose pCASP-3G124-HilA expression of a-GFP DARPin from Salmonella is disrupted pCASP-SptP1203m160-HilA triggers secretion of SptP120 fused to a-mCherry DARPin from Salmonella upon induction with arabinose pCASP-SptP120E3_5-HilA triggers secretion of SptP120 fused to E3_5 DARPin from Salmonella upon induction with arabinose pCASP-SptP120NS1-HilA triggers secretion of SptP120 fused to NS1 monobody from Salmonella upon induction with arabinose pCASP-SptP120K27-HilA triggers secretion of SptP120 fused to K27 DARPin from Salmonella upon induction with arabinose pCASP-SptP120K55-HilA triggers secretion of SptP120 fused to K55 DARPin from Salmonella upon induction with arabinose pCASP-SptP120-TDHilA triggers secretion of SptP120 fused to HA4-7c12 Tandem Monobody from Salmonella upon induction with arabinose pCASP-SptP92-TDHilA triggers secretion of SptP92 fused to HA4-7c12 Tandem Monobody from Salmonella upon induction with arabinose pCASP-SptP35-TDHilA triggers secretion of SptP35 fused to HA4-7c12 Tandem Monobody from Salmonella upon induction with arabinose pCASP-HilA empty vector triggers HilA overexpression upon induction with arabinose without any synthetic protein expressed from Salmonella pCMV R8.74 lentiviral packaging plasmid – gift from Didier Trono (Addgene plasmid # 22036) pMD2.G vesicular stomatitis G envelope expressing plasmid – gift from Dider Trono (Addgene plasmid # 12259) Lenti-AcGFP-Sec61IRES-Blast vector plasmid for integration and constitutive expression of AcGFP fused to Sec61 in mammalian cells – gift from D. Gerlich (IMBA, Vienna). .. Lenti-mCherry-WTKRAS-IRES-Blast vector plasmid for integration and constitutive expression of mCherry fused to wild-type KRAS in mammalian cells Lenti-mCherry-G12VKRAS-IRES-Blast vector plasmid for integration and constitutive expression of mCherry fused to KRASG12V in mammalian cells All plasmid maps and sequences can be found on Addgene.org Supplementary Table 4. .. List of cell lines used in this study Cell line Description HCT116 Human colorectal carcinoma derived cell line bearing a KRASG13D activating mutation – obtained from ATCC HCT116 G12V KRAS-mCherry HCT116 cells engineered to stably express mutated G12V KRAS fused to mCherry – created in this study by lentiviral transduction HCT116 Sec61-GFP HCT116 cells engineered to stably express Sec61 fused to AcGFP – created in this study by lentiviral transduction HeLa HeLa Kyoto – human cervix adenocarcinoma derived cell line – gift from the lab of D. Gerlich (IMBA, Vienna) HeLa Sec61-GFP HeLa Kyoto cells engineered to stably express Sec61 fused to AcGFP – gift from the lab of D. Gerlich (unpublished) HeLa H2B-GFP HeLa Kyoto cells engineered to stably express H2B fused to EGFP – gift from the lab of D. Gerlich ref. 6 HeLa alpha-Tubulin-mCherry HeLa Kyoto cells engineered to stably express alpha-Tubulin fused to mCherry – gift from the lab of D. Gerlich ref. 6 HeLa Sec61-GFP H2B-mCherry HeLa Kyoto cells engineered to stably express Sec61 fused to AcGFP and H2B fused to mCherry – created in this study by lentiviral transduction of HeLa Kyoto cells that also stably express H2B-mCherry.

    Article Title: Loss of MLL Induces Epigenetic Dysregulation of Rasgrf1 to Attenuate Kras-Driven Lung Tumorigenesis
    Article Snippet: .. Establishment of KrasG12V stably overexpressing cell line Lenti-mCherry-G12V-Kras plasmid was purchased from Addgene and used to generate stable cell lines. ..

    Clone Assay:

    Article Title: A novel, RAS-independent role for NF1 in microtubular dynamics and damage repair dictates sensitivity to T-DM1 in HER2-positive breast cancer
    Article Snippet: Images were processed using the ColonyArea ImageJ plugin250. .. KRASG12V from pDONR223_KRAS_p.G12V (Addgene #81665) was cloned on the lentiviral backbone pLenti CMV hygro dest (Addgene #17454). .. HEK293T cell transfection and infection of target cells was performed by Calcium Chloride followed by hygromycin B (InvivoGen) 0.5 mg/mL selection for 8 days.

    Generated:

    Article Title: Single-cell analyses reveal YAP/TAZ as regulators of stemness and cell plasticity in Glioblastoma
    Article Snippet: For constitutive expression of RTKs, cDNAs of HER2CA (from FUW-tetO-HER2-CA), PDGFRαCA (from pcDNA5FRT-EF-Pdgfrα-CA-EGFPN, Addgene#66789) and EGFRCA (from pBabe EGFR-L858R/T790M, Addgene#32073) were subcloned in CSII-CMV-MCS-IRES-puro empty vector backbone (obtained by substituting the blasticidin (bsd) resistance of CSII-CMV-MCS-IRES-bsd (a gift of H. Miyoshi) with the puromycin resistance). .. For constitutive lentiviral expression of mutant KRas, the pTomo KRasG12V was generated by subcloning the KRasG12V from the pBabe KRasG12V (Addgene#46746) to the pTomo-MCS Empty vector, obtained by substituting the loxP-RFP-loxP cassette from the pTomo vector (Addgene #26291) with a synthetic multiple cloning sites (MCS). .. For constitutive lentiviral expression of mutant KRas, the pTomo KRasG12V was generated by subcloning the KRasG12V from the pBabe KRasG12V (Addgene#46746) to the pTomo-MCS Empty vector, obtained by substituting the loxP-RFP-loxP cassette from the pTomo vector (Addgene #26291) with a synthetic multiple cloning sites (MCS).

    Subcloning:

    Article Title: Single-cell analyses reveal YAP/TAZ as regulators of stemness and cell plasticity in Glioblastoma
    Article Snippet: For constitutive expression of RTKs, cDNAs of HER2CA (from FUW-tetO-HER2-CA), PDGFRαCA (from pcDNA5FRT-EF-Pdgfrα-CA-EGFPN, Addgene#66789) and EGFRCA (from pBabe EGFR-L858R/T790M, Addgene#32073) were subcloned in CSII-CMV-MCS-IRES-puro empty vector backbone (obtained by substituting the blasticidin (bsd) resistance of CSII-CMV-MCS-IRES-bsd (a gift of H. Miyoshi) with the puromycin resistance). .. For constitutive lentiviral expression of mutant KRas, the pTomo KRasG12V was generated by subcloning the KRasG12V from the pBabe KRasG12V (Addgene#46746) to the pTomo-MCS Empty vector, obtained by substituting the loxP-RFP-loxP cassette from the pTomo vector (Addgene #26291) with a synthetic multiple cloning sites (MCS). .. For constitutive lentiviral expression of mutant KRas, the pTomo KRasG12V was generated by subcloning the KRasG12V from the pBabe KRasG12V (Addgene#46746) to the pTomo-MCS Empty vector, obtained by substituting the loxP-RFP-loxP cassette from the pTomo vector (Addgene #26291) with a synthetic multiple cloning sites (MCS).

    Cloning:

    Article Title: Single-cell analyses reveal YAP/TAZ as regulators of stemness and cell plasticity in Glioblastoma
    Article Snippet: For constitutive expression of RTKs, cDNAs of HER2CA (from FUW-tetO-HER2-CA), PDGFRαCA (from pcDNA5FRT-EF-Pdgfrα-CA-EGFPN, Addgene#66789) and EGFRCA (from pBabe EGFR-L858R/T790M, Addgene#32073) were subcloned in CSII-CMV-MCS-IRES-puro empty vector backbone (obtained by substituting the blasticidin (bsd) resistance of CSII-CMV-MCS-IRES-bsd (a gift of H. Miyoshi) with the puromycin resistance). .. For constitutive lentiviral expression of mutant KRas, the pTomo KRasG12V was generated by subcloning the KRasG12V from the pBabe KRasG12V (Addgene#46746) to the pTomo-MCS Empty vector, obtained by substituting the loxP-RFP-loxP cassette from the pTomo vector (Addgene #26291) with a synthetic multiple cloning sites (MCS). .. For constitutive lentiviral expression of mutant KRas, the pTomo KRasG12V was generated by subcloning the KRasG12V from the pBabe KRasG12V (Addgene#46746) to the pTomo-MCS Empty vector, obtained by substituting the loxP-RFP-loxP cassette from the pTomo vector (Addgene #26291) with a synthetic multiple cloning sites (MCS).



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    Image Search Results


    Fig. 1 The best- scored flexible docking poses of PER154A (A), PER160A (B), PER170A (C), and PER176A (D) docked to KRASG12V (PDB ID: 8AFD). Experi mental V12 and Q61 conformations have been shown in light gray, and results from flexible docking are shown in green. Hydrogen bonds are repre sented by dashed cyan lines

    Journal: BMC cancer

    Article Title: First molecules to reactivate RAS G12V GTPase activity.

    doi: 10.1186/s12885-025-13580-8

    Figure Lengend Snippet: Fig. 1 The best- scored flexible docking poses of PER154A (A), PER160A (B), PER170A (C), and PER176A (D) docked to KRASG12V (PDB ID: 8AFD). Experi mental V12 and Q61 conformations have been shown in light gray, and results from flexible docking are shown in green. Hydrogen bonds are repre sented by dashed cyan lines

    Article Snippet: Briefly, 500 ng of KRAS-WT or KRASG12V protein (both from SignalChem), 500 ng of GAP RT 01001-PER, molecules at selected concentrations – 10 μM, 1 μM DTT, 5 μM GDP and GTPase/GAP Buffer were applied to white non-transparent 96-well plates to a final volume of 25 μl per well.

    Techniques:

    Fig. 3 Analysis of the GTPase activity of the KRASG12V protein in the presence of selected molecules. (A) Verification of the action of GAP family proteins – a commercially available and self-obtained GAP RT 01001-PER on KRASWT and KRASG12V proteins. No functionality was observed for the commercial pro tein, while GAP RT 01001-PER showed GTPase activity for KRASWT and had no effect on KRASG12V. The GTPase activity of the KRASG12V protein was partially restored by selected molecules. These effects were observed both in the absence (B) and presence (C) of the GAP RT 01001-PER protein

    Journal: BMC cancer

    Article Title: First molecules to reactivate RAS G12V GTPase activity.

    doi: 10.1186/s12885-025-13580-8

    Figure Lengend Snippet: Fig. 3 Analysis of the GTPase activity of the KRASG12V protein in the presence of selected molecules. (A) Verification of the action of GAP family proteins – a commercially available and self-obtained GAP RT 01001-PER on KRASWT and KRASG12V proteins. No functionality was observed for the commercial pro tein, while GAP RT 01001-PER showed GTPase activity for KRASWT and had no effect on KRASG12V. The GTPase activity of the KRASG12V protein was partially restored by selected molecules. These effects were observed both in the absence (B) and presence (C) of the GAP RT 01001-PER protein

    Article Snippet: Briefly, 500 ng of KRAS-WT or KRASG12V protein (both from SignalChem), 500 ng of GAP RT 01001-PER, molecules at selected concentrations – 10 μM, 1 μM DTT, 5 μM GDP and GTPase/GAP Buffer were applied to white non-transparent 96-well plates to a final volume of 25 μl per well.

    Techniques: Activity Assay