Review

n terminal flag tag vector (Addgene inc)

Addgene inc is a verified supplier

About

News

Press Release

Team

Advisors

Partners

Contact

Bioz Stars

Bioz vStars

Buy from Supplier

Structured Review

Addgene inc n terminal flag tag vector

<t>BLNK</t> is a regulator of Met signaling (A) Time course signaling assay in response to HGF in <t>FLAG-BLNK-</t> and empty vector-transfected HEK293 cells is shown. Total Met (α-Met), BLNK (α-FLAG), phosphorylation of ERK (α-pERK) and total ERK (α-ERK) are represented. GAPDH (α-GAPDH) was used as loading control. (B) Time course signaling assay in response to HGF in NCIH2122 cells with siRNA-mediated knockdown of BLNK is shown. Total Met (α-Met), BLNK (α-BLNK), phosphorylation of ERK (α-pERK), total ERK (α-ERK), phosphorylation of AKT (α-pAKT) and total AKT (α-AKT) are represented. GAPDH (α-GAPDH) was used as loading control. (C) Time course signaling assay in response to HGF in NCIH1568 cells with siRNA-mediated knockdown of BLNK is shown. Total Met (α-Met), BLNK (α-BLNK), phosphorylation of ERK (α-pERK), total ERK (α-ERK), phosphorylation of AKT (α-pAKT) and total AKT (α-AKT) are represented. GAPDH (α-GAPDH) was used as loading control. (D)and(E) Phosphorylation of Met in response to HGF post siRNA-mediated knockdown of BLNK is assessed using immunoprecipitation of Met in NCIH2122 (D) and NCIH1568 (E). Total Met (α-Met), phosphorylated Met (α-pMet Y1234/Y1235) and BLNK (α-BLNK) are shown. GAPDH (α-GAPDH) was used as loading control.

N Terminal Flag Tag Vector, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/n terminal flag tag vector/product/Addgene inc
Average 93 stars, based on 2 article reviews

n terminal flag tag vector - by Bioz Stars, 2026-05

93/100 stars

Images

1) Product Images from "B cell linker protein (BLNK) is a regulator of Met receptor signaling and trafficking in non-small cell lung cancer"

Article Title: B cell linker protein (BLNK) is a regulator of Met receptor signaling and trafficking in non-small cell lung cancer

Journal: iScience

doi: 10.1016/j.isci.2022.105419

BLNK is a regulator of Met signaling (A) Time course signaling assay in response to HGF in FLAG-BLNK- and empty vector-transfected HEK293 cells is shown. Total Met (α-Met), BLNK (α-FLAG), phosphorylation of ERK (α-pERK) and total ERK (α-ERK) are represented. GAPDH (α-GAPDH) was used as loading control. (B) Time course signaling assay in response to HGF in NCIH2122 cells with siRNA-mediated knockdown of BLNK is shown. Total Met (α-Met), BLNK (α-BLNK), phosphorylation of ERK (α-pERK), total ERK (α-ERK), phosphorylation of AKT (α-pAKT) and total AKT (α-AKT) are represented. GAPDH (α-GAPDH) was used as loading control. (C) Time course signaling assay in response to HGF in NCIH1568 cells with siRNA-mediated knockdown of BLNK is shown. Total Met (α-Met), BLNK (α-BLNK), phosphorylation of ERK (α-pERK), total ERK (α-ERK), phosphorylation of AKT (α-pAKT) and total AKT (α-AKT) are represented. GAPDH (α-GAPDH) was used as loading control. (D)and(E) Phosphorylation of Met in response to HGF post siRNA-mediated knockdown of BLNK is assessed using immunoprecipitation of Met in NCIH2122 (D) and NCIH1568 (E). Total Met (α-Met), phosphorylated Met (α-pMet Y1234/Y1235) and BLNK (α-BLNK) are shown. GAPDH (α-GAPDH) was used as loading control.

Figure Legend Snippet: BLNK is a regulator of Met signaling (A) Time course signaling assay in response to HGF in FLAG-BLNK- and empty vector-transfected HEK293 cells is shown. Total Met (α-Met), BLNK (α-FLAG), phosphorylation of ERK (α-pERK) and total ERK (α-ERK) are represented. GAPDH (α-GAPDH) was used as loading control. (B) Time course signaling assay in response to HGF in NCIH2122 cells with siRNA-mediated knockdown of BLNK is shown. Total Met (α-Met), BLNK (α-BLNK), phosphorylation of ERK (α-pERK), total ERK (α-ERK), phosphorylation of AKT (α-pAKT) and total AKT (α-AKT) are represented. GAPDH (α-GAPDH) was used as loading control. (C) Time course signaling assay in response to HGF in NCIH1568 cells with siRNA-mediated knockdown of BLNK is shown. Total Met (α-Met), BLNK (α-BLNK), phosphorylation of ERK (α-pERK), total ERK (α-ERK), phosphorylation of AKT (α-pAKT) and total AKT (α-AKT) are represented. GAPDH (α-GAPDH) was used as loading control. (D)and(E) Phosphorylation of Met in response to HGF post siRNA-mediated knockdown of BLNK is assessed using immunoprecipitation of Met in NCIH2122 (D) and NCIH1568 (E). Total Met (α-Met), phosphorylated Met (α-pMet Y1234/Y1235) and BLNK (α-BLNK) are shown. GAPDH (α-GAPDH) was used as loading control.

Techniques Used: Plasmid Preparation, Transfection, Immunoprecipitation

$Met-GRB2 interaction is increased in the presence of BLNK, and BLNK-GRB2 interaction is increased in the presence of active Met (A) (Left panel) Schematic of modified MaMTH assay is shown. Created with Biorender.com . (Right panel) MaMTH assay examining the interaction between Met and GRB2 or PEX7 (negative control) in the presence of GFP-tagged BLNK or GFP-tagged LCP2 is shown. Significance was assessed using unpaired two-tailed Welch’s t test. Results are shown as mean ± SD, with n = 4 technical replicates. ∗∗∗p<0.001. Protein expression of all constructs was detected using western blotting. Met bait (α-V5), GRB2/NCK2 preys (α-FLAG) and BLNK/LCP2 (α-GFP) are shown. GAPDH (α-GAPDH) was used as a loading control. (B) Co-immunoprecipitation investigating interaction between GRB2 and BLNK in the presence of Met under different conditions is shown. Phosphorylated Met was detected using α-pY antibody. Tubulin (α-Tubulin) was used as a loading control. IP-Immunoprecipitated fraction; WCL – Whole cell lysate. (C) SIMPL-ELISA assay examining interaction between GRB2 bait and various BLNK mutants or PEX7 (negative control) preys is shown. Mock - no transfection, empty - empty vector transfection. The spliced signal is expressed as relative luciferase units and is normalized to bait and prey expression. Grubbs test was performed to remove any outliers. Significance was assessed using unpaired two-tailed Student’s t test. Results are shown as mean ± SD, with n= minimum of 2 technical replicates after Grubbs test. ∗p<0.05; ∗∗p<0.01; ∗∗∗p<0.001. (D) Time course signaling assay in response to HGF, in the presence of BLNK wildtype, BLNK P204A212AY72F or empty vector in HEK293 cells is shown. Total Met (α-Met), BLNK (α-FLAG), phosphorylation of ERK (α-pERK) and total ERK (α-ERK) are represented. GAPDH (α-GAPDH) was used as loading control.$
Figure Legend Snippet: Met-GRB2 interaction is increased in the presence of BLNK, and BLNK-GRB2 interaction is increased in the presence of active Met (A) (Left panel) Schematic of modified MaMTH assay is shown. Created with Biorender.com . (Right panel) MaMTH assay examining the interaction between Met and GRB2 or PEX7 (negative control) in the presence of GFP-tagged BLNK or GFP-tagged LCP2 is shown. Significance was assessed using unpaired two-tailed Welch’s t test. Results are shown as mean ± SD, with n = 4 technical replicates. ∗∗∗p<0.001. Protein expression of all constructs was detected using western blotting. Met bait (α-V5), GRB2/NCK2 preys (α-FLAG) and BLNK/LCP2 (α-GFP) are shown. GAPDH (α-GAPDH) was used as a loading control. (B) Co-immunoprecipitation investigating interaction between GRB2 and BLNK in the presence of Met under different conditions is shown. Phosphorylated Met was detected using α-pY antibody. Tubulin (α-Tubulin) was used as a loading control. IP-Immunoprecipitated fraction; WCL – Whole cell lysate. (C) SIMPL-ELISA assay examining interaction between GRB2 bait and various BLNK mutants or PEX7 (negative control) preys is shown. Mock - no transfection, empty - empty vector transfection. The spliced signal is expressed as relative luciferase units and is normalized to bait and prey expression. Grubbs test was performed to remove any outliers. Significance was assessed using unpaired two-tailed Student’s t test. Results are shown as mean ± SD, with n= minimum of 2 technical replicates after Grubbs test. ∗p<0.05; ∗∗p<0.01; ∗∗∗p<0.001. (D) Time course signaling assay in response to HGF, in the presence of BLNK wildtype, BLNK P204A212AY72F or empty vector in HEK293 cells is shown. Total Met (α-Met), BLNK (α-FLAG), phosphorylation of ERK (α-pERK) and total ERK (α-ERK) are represented. GAPDH (α-GAPDH) was used as loading control.

Techniques Used: Modification, Negative Control, Two Tailed Test, Expressing, Construct, Western Blot, Immunoprecipitation, Enzyme-linked Immunosorbent Assay, Transfection, Plasmid Preparation, Luciferase

Figure Legend Snippet:

Techniques Used: Enzyme-linked Immunosorbent Assay, Recombinant, Transfection, Western Blot, CRISPR, Knock-Out, Sequencing, Software

Image Search Results

Journal: iScience

Article Title: B cell linker protein (BLNK) is a regulator of Met receptor signaling and trafficking in non-small cell lung cancer

doi: 10.1016/j.isci.2022.105419

Figure Lengend Snippet: BLNK is a regulator of Met signaling (A) Time course signaling assay in response to HGF in FLAG-BLNK- and empty vector-transfected HEK293 cells is shown. Total Met (α-Met), BLNK (α-FLAG), phosphorylation of ERK (α-pERK) and total ERK (α-ERK) are represented. GAPDH (α-GAPDH) was used as loading control. (B) Time course signaling assay in response to HGF in NCIH2122 cells with siRNA-mediated knockdown of BLNK is shown. Total Met (α-Met), BLNK (α-BLNK), phosphorylation of ERK (α-pERK), total ERK (α-ERK), phosphorylation of AKT (α-pAKT) and total AKT (α-AKT) are represented. GAPDH (α-GAPDH) was used as loading control. (C) Time course signaling assay in response to HGF in NCIH1568 cells with siRNA-mediated knockdown of BLNK is shown. Total Met (α-Met), BLNK (α-BLNK), phosphorylation of ERK (α-pERK), total ERK (α-ERK), phosphorylation of AKT (α-pAKT) and total AKT (α-AKT) are represented. GAPDH (α-GAPDH) was used as loading control. (D)and(E) Phosphorylation of Met in response to HGF post siRNA-mediated knockdown of BLNK is assessed using immunoprecipitation of Met in NCIH2122 (D) and NCIH1568 (E). Total Met (α-Met), phosphorylated Met (α-pMet Y1234/Y1235) and BLNK (α-BLNK) are shown. GAPDH (α-GAPDH) was used as loading control.

Article Snippet: For signaling and immunoprecipitation of BLNK in HEK293 cells, N terminally FLAG-tagged BLNK was generated by cloning BLNK entry plasmid (Source: Human ORFeome V8.1) into pCMV5-based N terminal FLAG tag vector (Addgene).

Techniques: Plasmid Preparation, Transfection, Immunoprecipitation

Journal: iScience

Article Title: B cell linker protein (BLNK) is a regulator of Met receptor signaling and trafficking in non-small cell lung cancer

doi: 10.1016/j.isci.2022.105419

Figure Lengend Snippet: Met-GRB2 interaction is increased in the presence of BLNK, and BLNK-GRB2 interaction is increased in the presence of active Met (A) (Left panel) Schematic of modified MaMTH assay is shown. Created with Biorender.com . (Right panel) MaMTH assay examining the interaction between Met and GRB2 or PEX7 (negative control) in the presence of GFP-tagged BLNK or GFP-tagged LCP2 is shown. Significance was assessed using unpaired two-tailed Welch’s t test. Results are shown as mean ± SD, with n = 4 technical replicates. ∗∗∗p<0.001. Protein expression of all constructs was detected using western blotting. Met bait (α-V5), GRB2/NCK2 preys (α-FLAG) and BLNK/LCP2 (α-GFP) are shown. GAPDH (α-GAPDH) was used as a loading control. (B) Co-immunoprecipitation investigating interaction between GRB2 and BLNK in the presence of Met under different conditions is shown. Phosphorylated Met was detected using α-pY antibody. Tubulin (α-Tubulin) was used as a loading control. IP-Immunoprecipitated fraction; WCL – Whole cell lysate. (C) SIMPL-ELISA assay examining interaction between GRB2 bait and various BLNK mutants or PEX7 (negative control) preys is shown. Mock - no transfection, empty - empty vector transfection. The spliced signal is expressed as relative luciferase units and is normalized to bait and prey expression. Grubbs test was performed to remove any outliers. Significance was assessed using unpaired two-tailed Student’s t test. Results are shown as mean ± SD, with n= minimum of 2 technical replicates after Grubbs test. ∗p<0.05; ∗∗p<0.01; ∗∗∗p<0.001. (D) Time course signaling assay in response to HGF, in the presence of BLNK wildtype, BLNK P204A212AY72F or empty vector in HEK293 cells is shown. Total Met (α-Met), BLNK (α-FLAG), phosphorylation of ERK (α-pERK) and total ERK (α-ERK) are represented. GAPDH (α-GAPDH) was used as loading control.

Techniques: Modification, Negative Control, Two Tailed Test, Expressing, Construct, Western Blot, Immunoprecipitation, Enzyme-linked Immunosorbent Assay, Transfection, Plasmid Preparation, Luciferase

Journal: iScience

Article Title: B cell linker protein (BLNK) is a regulator of Met receptor signaling and trafficking in non-small cell lung cancer

doi: 10.1016/j.isci.2022.105419

Figure Lengend Snippet:

Techniques: Enzyme-linked Immunosorbent Assay, Recombinant, Transfection, Western Blot, CRISPR, Knock-Out, Sequencing, Software

Review

Structured Review

Images

1) Product Images from "B cell linker protein (BLNK) is a regulator of Met receptor signaling and trafficking in non-small cell lung cancer"

Similar Products

Image Search Results