Invitrogen

V5 Tag Monoclonal Antibody (TCM5), eBioscience™

高级验证

This Antibody was verified by Cell treatment to ensure that the antibody binds to the antigen stated.

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V5 Tag Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescent analysis of V5 Tag was performed using 70% confluent V5-H3-6XHis transfected HEK-293 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with V5 Tag Monoclonal Antibody (TCM5), eBioscience™ (Product # 14-6796-82) at 5 µg/mL and Histone H3 Recombinant Polyclonal Antibody (Product # 711055) at 0.5 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 647 (Product # A32728) and Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32731) respectively at a dilution of 1:2000 for 45 minutes at room temperature. Panel a (Nuclei: Red) represents V5 Tag. Panel b (Nuclei: Green) represents Histone H3. Panel c (Nuclei: Blue) represents ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). Panel d represents the merged image showing the co-localization of nuclear signals in transfected cells. Panel e represents untransfected HEK-293 cells. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

V5 Tag Antibody in Western Blot (WB) — Western blot was performed using Anti-V5 Mouse Monoclonal Antibody (Product # 14-6796-82) by loading whole cell extracts of untransfected and transiently transfected HEK-293E lysates: untransfected, 50 µg (Lane 1), empty vector control, 50 µg (Lane 2), V5-H3-His, 50 µg (Lane 3), Myc-p65-V5, 50 µg (Lane 4), Myc-p65-V5, 25 µg (Lane 5), Myc-p65-V5, 12.5 µg (Lane 6), Myc-p65-V5, 6.26 µg (Lane 7), FLAG-P65-HA, 50 µg (Lane 8) and 25 ng of Positope (Lane 9) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). A ~19 kDa band corresponding to V5-H3-His and ~65 kDa band corresponding to Myc-p65-V5 were observed in HEK293E transfected lysates on probing with the primary antibody (1 µg/mL) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1500 (Product # A44241). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Positope (Product # R90050) is a 53 kDa recombinant protein consisting multiple epitope tags along with V5 tag, which has been used as a positive control for V5 tag detection. No cross-reactivity was seen with FLAG-tagged p65 expressing lysate.

V5 Tag Antibody in Western Blot, Immunocytochemistry (WB, ICC/IF) — Fig. 6. Delivery of glioma-relevant mutations into genetically normal human NSCs. (A) Schematic depiction of the targeting strategy to knock out TP53 via gene targeting. CRISPR sgRNA pair targeting site is indicated with a yellow triangle. Horizontal arrows indicate PCR genotyping primers for assaying TP53 locus targeting (PCR1 and PCR2) and presence of an indel within the second allele (PCR3). (B) Representative phase contrast images of puromycin-resistant human NSC colony after 10 days of selection. Right-hand image is a magnification of the boxed area on the left. Scale bar: 100 um. (C) PCR-based genotyping (top) and WB analysis (bottom) of human TP53 targeted NSC clonal lines. Parental, non-transfected cells were used as a control. (D) Sanger sequencing trace of an exemplar correctly targeted clone harbouring an 85 bp deletion on the second allele. (E) Growth curve analysis of the TP53 targeted clones harbouring the 85 bp deletion confirming the positive proliferative effect of p53 ablation. (F) Illustration of the strategy for gene targeting the first H3F3A coding exon. Yellow triangle indicates sgRNA pair targeting site. Horizontal arrows indicate PCR genotyping primers. (G) ICC using a V5 tag-specific antibody to identify targeted cells (arrowheads). DAPI was used for nuclear staining. Scale bar: 50 um. (H) Western blotting using V5 tag and histone H3-specific antibodies and PCR genotyping of parental and transfected cells. (I) Sanger sequencing traces of the genotypin

V5 Tag Antibody in Immunohistochemistry (IHC) — Published figure using V5 Tag monoclonal antibody (Product # 14-6796-82) in Immunohistochemistry

V5 Tag Antibody in Western Blot, Immunoprecipitation (WB, IP) — Figure 6 G34R mutation precludes binding to the transcriptional repressor ZMYND11 (A) Immunoblotting of immunoprecipitated H3.3-WT and H3.3-G34R ectopically expressed in G34R-KO pGBM002 cells. Endogenous (lower band) and V5-tagged ectopic H3.3 (upper band) are indicated. Loss of K36me3 is specific to ectopic H3.3-G34R. Non-transfected G34R-KO cells were used as controls (first and fourth lanes). (B) Metagene plot (left) and boxplot (right) showing H3.3-G34R coverage for expressed, upregulated, downregulated - all, and downregulated - forebrain associated genes. Expressed genes were defined as those with mean RPKM > 1 within pGBM002 parental samples (see STAR methods ). All other gene sets were defined on the basis of differential expression between G34R-KO and parental cells ( Figures 4 I-4L). *** p < 0.005 and **** p < 0.0001 for pairwise comparisons against the ""all expressed"" group. See STAR methods for details on the statistical test. (C) Genomic track showing H3.3-G34R enrichment and mRNA expression at DMRTA2 (left) and DLX1 (right) loci. DMRTA2 and DLX2 are shown, respectively, as representative downregulated and unchanged forebrain-related genes. (D) Volcano plot showing differentially bound partners of ectopically expressed WT and G34R H3.3 identified through IP-MS. x axis shows the fold-change value (G34R/WT) and y axis shows p value (-log 10 scale) for statistical significance. Absolute 3-fold change and a p value of 0.1 (vertical and horizontal dashed lines, resp

V5 Tag Antibody — Antibody specificity was demonstrated by detection of different targets fused to V5 tag in transiently transfected lysates tested. Relative detection of V5 tag was observed across different proteins fused with V5 tag in V5-H3-His (Lane 3) and Myc-p65-V5 (Lane 4-7), using V5 Tag Monoclonal Antibody (TCM5), eBioscience™ (Product # 14-6796-82) in Western Blot. This product has been shown to detect DYKDDDDK Tag at both N- and C- termini of a fusion protein. {RE}

产品信息

14-6796-82

应用	建议稀释比	已发表文章
免疫印迹 (WB)	1:1,000	查看4篇已发表文章 4篇已发表文章
免疫组化 (IHC)	-	查看2篇已发表文章 2篇已发表文章
免疫细胞化学 (ICC/IF)	1:100	查看4篇已发表文章 4篇已发表文章
免疫沉淀 (IP)	-	查看1篇已发表文章 1篇已发表文章

产品规格
种属反应	Tag
已发表种属	Tag
宿主/亚型	Mouse / IgG2b, kappa
分类	Monoclonal
类型	Antibody
克隆号	TCM5
偶联物	Unconjugated PE PE-Cyanine7
形式	Liquid
浓度	0.5 mg/mL
纯化类型	Affinity chromatography
保存液	PBS, pH 7.2
内含物	0.09% sodium azide
保存条件	4° C
RRID	AB_10718239

产品详细信息

Description: This TCM5 monoclonal antibody recognizes recombinant proteins containing a V5 epitope tag. The V5 epitope is naturally present on the P and V proteins of the paramyxovirus, SV5. TCM5 specifically recognizes the sequence GKPIPNPLLGLDST.

Applications Reported: This TCM5 antibody has been reported for use in western blotting, and immunocytochemistry.

Applications Tested: This TCM5 antibody has been tested by western blot and immunofluorescent staining of V5-transfected cells and can be used at less than or equal to 1 µg/mL. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

Purity: Greater than 90%, as determined by SDS-PAGE.

Aggregation: Less than 10%, as determined by HPLC.

Filtration: 0.2 µm post-manufacturing filtered.

靶标信息

Epitope tags provide a method to localize gene products in a variety of cell types, study the topology of proteins and protein complexes, identify associated proteins, and characterize newly identified, low abundance or poorly immunogenic proteins when protein specific antibodies are not available. The V5 epitope tag is derived from a small epitope (Pk) present on the P and V proteins of the paramyxovirus of simian virus (SV5). The V5 tag is usually used with all 14 amino acids (GKPIPNPLLGLDST), although it has also been used with a shorter 9 amino acid sequence (IPNPLLGLD).

仅用于科研。不用于诊断过程。未经明确授权不得转售。

Product References

Regional identity of human neural stem cells determines oncogenic responses to histone H3.3 mutants.

Cell stem cell

Bressan RB,Southgate B,Ferguson KM,Blin C,Grant V,Alfazema N,Wills JC,Marques-Torrejon MA,Morrison GM,Ashmore J,Robertson F,Williams CAC,Bradley L,von Kriegsheim A,Anderson RA,Tomlinson SR,Pollard SM

Published figure using V5 Tag monoclonal antibody (Product # 14-6796-82) in Immunoprecipitation

Thu May 06 00:00:00 EDT 2021

Regional identity of human neural stem cells determines oncogenic responses to histone H3.3 mutants.

Cell stem cell

Bressan RB,Southgate B,Ferguson KM,Blin C,Grant V,Alfazema N,Wills JC,Marques-Torrejon MA,Morrison GM,Ashmore J,Robertson F,Williams CAC,Bradley L,von Kriegsheim A,Anderson RA,Tomlinson SR,Pollard SM

Published figure using V5 Tag monoclonal antibody (Product # 14-6796-82) in Immunoprecipitation

Thu May 06 00:00:00 EDT 2021

Calcium-Mediated Liposome Fusion to Engineer Giant Lipid Vesicles with Cytosolic Proteins and Reconstituted Mammalian Proteins.

Advanced biosystems

Schmid YRF,Scheller L,Buchmann S,Dittrich PS

Published figure using V5 Tag monoclonal antibody (Product # 14-6796-82) in Immunocytochemistry

Sun Nov 01 00:00:00 EDT 2020

Endogenous epitope-tagging of Tet1, Tet2 and Tet3 identifies TET2 as a naïve pluripotency marker.

Life science alliance

Pantier R,Tatar T,Colby D,Chambers I

14-6796 was used in Western Blotting to show that TET2 binds to the transcription factor NANOG.

Tue Oct 01 00:00:00 EDT 2019

Endogenous epitope-tagging of Tet1, Tet2 and Tet3 identifies TET2 as a naïve pluripotency marker.

Life science alliance

Pantier R,Tatar T,Colby D,Chambers I

14-6796 was used in Western Blotting to show that TET2 binds to the transcription factor NANOG.

Tue Oct 01 00:00:00 EDT 2019

The Effect of Selective c-MET Inhibitor on Hepatocellular Carcinoma in the MET-Active, β-Catenin-Mutated Mouse Model.

Gene expression

Zhan N,Michael AA,Wu K,Zeng G,Bell A,Tao J,Monga SP

14-6796 was used in Immunohistochemistry to investigate the effect of EMD1214063 on hepatocellular carcinoma in the MET-Active, Β-catenin-mutated mouse model.

Fri May 18 00:00:00 EDT 2018

An efficient and scalable pipeline for epitope tagging in mammalian stem cells using Cas9 ribonucleoprotein.

eLife

Dewari PS,Southgate B,Mccarten K,Monogarov G,O'Duibhir E,Quinn N,Tyrer A,Leitner MC,Plumb C,Kalantzaki M,Blin C,Finch R,Bressan RB,Morrison G,Jacobi AM,Behlke MA,von Kriegsheim A,Tomlinson S,Krijgsveld J,Pollard SM

14-6796 was used in Immunocytochemistry-immunoflourescence to demonstrate improved CRISPR knock-in efficiencies using co-delivery of Cas9 protein pre-complexed with two-part synthetic modified RNAs and single-stranded oligodeoxynucleotide repair templates.

Wed Apr 11 00:00:00 EDT 2018

Disruptions in asymmetric centrosome inheritance and WDR62-Aurora kinase B interactions in primary microcephaly.

Scientific reports

Sgourdou P,Mishra-Gorur K,Saotome I,Henagariu O,Tuysuz B,Campos C,Ishigame K,Giannikou K,Quon JL,Sestan N,Caglayan AO,Gunel M,Louvi A

14-6796 was used in Immunocytochemistry to study the critical and diverse functions of WDR62 in neocortical development, and the mechanisms by which its disruption leads to a plethora of structural abnormalities.

Wed Mar 08 00:00:00 EST 2017

Disruptions in asymmetric centrosome inheritance and WDR62-Aurora kinase B interactions in primary microcephaly.

Scientific reports

Sgourdou P,Mishra-Gorur K,Saotome I,Henagariu O,Tuysuz B,Campos C,Ishigame K,Giannikou K,Quon JL,Sestan N,Caglayan AO,Gunel M,Louvi A

Wed Mar 08 00:00:00 EST 2017

Efficient CRISPR/Cas9-assisted gene targeting enables rapid and precise genetic manipulation of mammalian neural stem cells.

Development (Cambridge, England)

Bressan RB,Dewari PS,Kalantzaki M,Gangoso E,Matjusaitis M,Garcia-Diaz C,Blin C,Grant V,Bulstrode H,Gogolok S,Skarnes WC,Pollard SM

Published figure using V5 Tag monoclonal antibody (Product # 14-6796-82) in Immunocytochemistry

Wed Feb 15 00:00:00 EST 2017

Efficient CRISPR/Cas9-assisted gene targeting enables rapid and precise genetic manipulation of mammalian neural stem cells.

Development (Cambridge, England)

Bressan RB,Dewari PS,Kalantzaki M,Gangoso E,Matjusaitis M,Garcia-Diaz C,Blin C,Grant V,Bulstrode H,Gogolok S,Skarnes WC,Pollard SM

Published figure using V5 Tag monoclonal antibody (Product # 14-6796-82) in Immunocytochemistry

Wed Feb 15 00:00:00 EST 2017

生物信息学

蛋白别名: simian virus 5 antibody; SPV5gp2; sv5; v-5; V5; V5 Epitope Tag

V5 Tag Monoclonal Antibody (TCM5), eBioscience™

产品信息

14-6796-82

免疫印迹 (WB)

免疫组化 (IHC)

免疫细胞化学 (ICC/IF)

免疫沉淀 (IP)

种属反应

已发表种属

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分类

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形式

浓度

纯化类型

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内含物

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