TCF1/TCF7 (C63D9) Rabbit mAb (Alexa Fluor ® 647 Conjugate) 6709S from Cell Signaling Technology

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Cancer Immunology, Immunotherapy
December 01 2022

Fig 1: Tcf1Tg TCR-engineered CD8+ T cells are reinforced against apoptosis A Persistence of TCR-engineered T cells over the course of in vitro culture with SK37 without exogenous IL-2. CD8+ T cells were engineered to express the 19,305-TCR and Tcf-1 and were then seeded onto SK37 monolayers without IL-2 supplementation. After (72, 196, 336) hours T cells were collected with uniform technique and were assayed via flow-cytometry-based detection of CD45 + cells with forward-scatter and side-scatter properties typical of lymphocytes, as seen in Supplemental Fig. 3A. CountBrite Absolute Counting Beads were used to ascertain cell numbers. A t-test was used to compare cell numbers at each timepoint, with an asterisk indicative of a comparisons that yielded a p-value less than 0.05. B Viability of TCR-engineered T cells during extended co-culture with SK37. T cells were labelled with annexin-v after 196-h of co-culture. A gate captures cells which labelled positive for annexin-v. CD45 + GFP + Zombie- cells with lymphocyte forward-scatter and side-scatter profiles were assayed. To the right is a bar graph comparing frequencies of apoptotic cells. A t-test was applied to assess statistical significance, without pairing. N = 3 clones propagated from the same donor. Asterisks indicate comparisons which yielded a p-value less than 0.05. C Mitochondria polarization of T cells during extended co-culture with SK37, as assessed via labelling with MitoTrackerDeepRedFM [10 nM] after 196-h. To the left are flow-cytometry histograms for T cells transduced to express Tcf-1 (red) or GFP (black) (along with the 19,305-TCR), with median fluorescence intensity listed underneath. Gates (seen in Supplemental Fig. 3B) allowed detection of MitoTracker in viable cells (with CD45 + GFP + Zombie- profiles). To the right is a bar graph. A t-test was applied to assess statistical significance, without pairing, with asterisks reflective of comparisons which yielded a p-value less than 0.05. N = 3 clones from the same donor. D Cell-cycle activity of T cells cultured with SK37, as influenced by IL-2. T cells were assayed for Ki67 content 48-h after initiation of co-culture, with or without IL-2 supplemented media (300 IU/ml). Gates capture cells which had intermediate and high labelling for Ki67. CD45 + GFP + CD8+ T cells were assayed, as seen in Supplemental Fig. 3C. To the right is a graph comparing frequencies of Ki67 + cells. t-tests were applied without pairing, for N = 3 clones from the same donor. Asterisks indicate comparisons which yielded a p-value less than 0.05. E MitoTrackerDeepRedFM labelling [10 nM] in relation to CD8 co-receptor labelling, in T cells co-cultured for 48-h with SK37. Gates were applied to observe CD45 + GFP + cells which did not uptake viability dye. To the right is a bar graph comparing MitoTracker labelling. t-tests were used to assess significance, without pairing. N = 3 clones from the same donor. Asterisks indicate comparisons which yielded a p-value less than 0.05. F Viability after 48-h of co-culture. This graph compares the annexin-v labelling of Tcf-1 transgenic TCR-engineered CD8+ T cells to GFP transgenic TCR-engineered CD8+ T cells. It compares CD45 + GFP + CD8+ T cells after 48 h of co-culture with SK37. A t-test was used to assess significance, without pairing. N = 3 clones from the same donor. Asterisks are indicative of comparisons which yielded a p-value of less than 0.05
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Nature immunology
September 01 2021

Fig 2: TCF-1 deficiency selectively reprograms Treg-cells without compromising their core signature.Treg-cells were isolated from the mesenteric lymph nodes of Foxp3Cre and Tcf7fl/flFoxp3Cre mice. (a) Representative FACS histograms of MLN purified cells from Foxp3Cre Tcf7fl/fl and control Foxp3Cre showing selective loss of TCF-1 from Treg-cells in Foxp3Cre Tcf7fl/fl mice. (b and c) Histogram plots showing the cumulative data of the same. (b: n = 4; p < 0.0001 & c: n = 5) Data are representative of two independent experiments and n represents biologically independent replicate mice; means ± SEM; two-sided, unpaired t-test. (d) GSEA plot comparing the enrichment of genes expressed more strongly in Foxp3Cre versus Foxp3CreTcf7fl/fl Treg-cells.
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Nature immunology
September 01 2021

Fig 3: Treg-cells are activated and polarized during polyposis.Treg_cells were isolated from the mesenteric lymph nodes of WT and APC?486 mice. (a) UMAP projection (left panel) and fraction of cells in each cell type (stack bars; right panel) for APC?486 and control B6 Treg-cells. Data are from two replicates. (b) Dot plot showing the expression of Tcf7 across all cell types in Apc?486 and control B6 Treg-cells. Color and size of the dots are proportional to the expression level and percent of cells expressing Tcf7 in each indicated cluster. (c) Expression of Socs3, Jund, Lag3 and Maf between APC?486 and B6 cells projected on the UMAP. See TableS4 for the full list. The fold change in percent of cells expressing the indicated gene in each cell type is proportional to the circle size. Adjusted-p-values > 0.01 are masked with gray color. (d) Expression changes of the most differentially expressed genes between APC?486 and control B6 Treg-cells. See TableS4 for the full list. The fold change in expression intensities is color-coded. (e) RNA velocity vectors overlaid on UMAP for B6 (left) and APC?486 (right) Treg-cells.
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Nature communications
November 24 2021

Fig 4: LSD1 loss unleashes TCF1-controlled gene expression in CD8+ TILs.a A volcano plot showing differential gene expression by LSD1-deficient versus wildtype CD8+CD44+PD-1+ TILs of MC38 tumors (n = 4 per group, FC > 1.5 and FDR < 0.01 as the cutoff). Non-gray colored dots mean significant difference. b Percentages of genes bound by TCF1 among the differentially expressed genes in LSD1-deficient versus wildtype CD8+ TILs or among total genes in genome. c GSEA analysis in Lsd1 KO versus WT CD8+ TILs (n = 4 per group). Gene list was ranked with signed (from log2FC) likelihood ratio from Lsd1 KO versus WT comparison. Kolmogorov–Smirnov test was used to determine the enrichment score. d, e A representative flow plot of IL7Ra expression (d) and IL7Ra MFI (e) of MC38 tumor-infiltrating CD8+PD-1+ T cells (n = 4 per group). f, g A representative flow plot of Slamf6 expression (f) and Slamf6 MFI (g) of MC38 tumor-infiltrating CD8+PD-1+ T cells (n = 4 per group). Data represent two independent experiments (d–g) and are presented as mean ± SEM (e, g). Statistical significance was determined by two-sided unpaired t test (e, g). Source data are provided as a Source data file.
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Nature communications
November 24 2021

Fig 5: LSD1 depletion preserves intratumoral TCF1+PD-1int progenitor exhausted CD8+ T cells.a, b Frequencies (a) and PD-1 MFI (b) of CD8+PD-1+ TILs on day 18 after MC38 tumor implantation (Lsd1f/f group, n = 4; Cd4-Cre+Lsd1f/f group, n = 5). c–e Representative flow plots (c), percentages (d) and cell numbers (e) of TCF1+PD-1int, TCF1+PD-1hi and TCF1-PD-1hi cells in CD8+ TILs analyzed by flow cytometry on day 12 and 18 after MC38 tumor implantation (Lsd1f/f d12 group, n = 5; Cd4-Cre+Lsd1f/f d12 group, n = 5; Lsd1f/f d18 group, n = 4; Cd4-Cre+Lsd1f/f d18 group, n = 4). f Percentages of TCF1+PD-1int, TCF1+PD-1hi and TCF1-PD-1hi cells in CD8+Tetramer+ TILs on day 18 after MC38-OVA tumor implantation (Lsd1f/f group, n = 14; Cd4-Cre+Lsd1f/f group, n = 10). g, h, Slamf6 (g) and Tim3 (h) expression by different subpopulations of CD8+ TILs analyzed by flow cytometry (Lsd1f/f group, n = 8; Cd4-Cre+Lsd1f/f group, n = 6). i, j, Representative flow plots of Ly6C expression (i) and percentages of Ly6C+ cells (j) in different subpopulations of CD8+ TILs (n = 4 per group). k, l GzmB (k) and Ki-67 (l) expression by different subpopulations of CD8+ TILs analyzed by flow cytometry on day 18 (n = 4 per group). m, n Cell apoptosis analysis by Annexin V staining (m) and active Casp3/7 staining (n) of CD8+ TILs isolated from MC38 tumors on day 14 (n = 5 per group). Data represent two independent experiments (a–e, g–n) or are pooled from three independent experiments (f) and are presented as mean ± SEM (a–n). Statistical significance was determined by two-sided unpaired t test (a–n). Source data are provided as a Source data file.

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Cell Signaling Technology for
TCF1/TCF7 (C63D9) Rabbit mAb (Alexa Fluor ® 647 Conjugate)

Cell Signaling Technology's TCF1/TCF7 (C63D9) Rabbit mAb (Alexa Fluor ® 647 Conjugate) is a Rabbit monoclonal antibody. The TCF1/TCF7 (C63D9) Rabbit mAb (Alexa Fluor ® 647 Conjugate) Antibody was generated using TCF7 as the antigen. It reacts with Homo Sapiens, Human, and Mouse. This antibody has been shown to work in applications such as: Flow Cytometry.

Description

TCF1/TCF7 (C63D9) Rabbit mAb (Alexa Fluor® 647 Conjugate) detects endogenous levels of total TCF1/TCF7 protein. This antibody does not recognize the dominant negative isoforms of TCF1/TCF7 lacking the amino-terminal β-catenin binding domain and does not cross-react with LEF1

Product Specs
ItemTCF1/TCF7 (C63D9) Rabbit mAb (Alexa Fluor ® 647 Conjugate)
CompanyCell Signaling Technology
Price Pricing Info Supplier Page View Company Product Page
Catalog Number6709S
Quantity100 µl
ApplicationsFC-FP
Conjugate/TagAlexa Fluor ® 647
ReactivityH, M
TargetTCF7
HostRabbit
NCBI Full Gene Nametranscription factor 7
NCBI Gene AliasesTCF-1
ClonalityMonoclonal
Research AreaDevelopmental Biology
CloneC63D9
Molecular Weight48, 50

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(1) IL-15 boosts activated HBV core-specific CD8+ progenitor cells via metabolic rebalancing in persistent HBV infectioniScienceJanuary 19, 2024Julia Peña-Asensio, Henar Calvo-Sánchez, Joaquín Miquel, Eduardo Sanz-de-Villalobos, Alejandro González-Praetorius, Miguel Torralba, Juan-Ramón LarrubiaClone RPA-T8" anti-human TCF1/TCF7 mAb (Alexa Fluor® 647 conjugate),Cell Signaling Technology,"Cat # 6709 RRID:AB_2797631, Clone C63D9" anti-human CX3CR1 mAb (FITC),BioLegend,"Cat # 341605, RRID:AB_1626276,

(2) Single-cell CRISPR screens in vivo map T cell fate regulomes in cancerNatureNovember 15, 2023Peipei Zhou, Hao Shi, Hongling Huang, Xiang Sun, Sujing Yuan, Nicole M. Chapman, Jon P. Connelly, Seon Ah Lim, Jordy Saravia, Anil KC, et al.IC3765A, 1:100) was from R&D Systems, and anti-RBPJ (D10A4, 5313 T), Alexa Fluor 647–anti-TCF-1 (C63D9, 6709, 1:100) and APC–anti-pS6 (S235/236) (D57.2.2E, 14733, 1:100) were from Cell Signaling Technology. To

(3) Overcoming lung cancer immunotherapy resistance by combining nontoxic variants of IL-12 and IL-2JCI InsightOctober 9, 2023Brendan L. Horton, Alicia D. D’Souza, Maria Zagorulya, Chloe V. McCreery, Gita C. Abhiraman, Lora Picton, Allison Sheen, Yash Agarwal, Noor Momin, K. Dane Wittrup, et al.conjugated to PE-Cy7 (BioLegend 109110); anti–TCF-1 clone C63D9 conjugated to AF647 (Cell Signaling 6709S) or AF488 (Cell Signaling 6444S); and anti–TIM-3 clone RMT3-23 conjugated to BV421 (BioLegend 119723). IL12-MSA

(4) Sex-Biased T-cell Exhaustion Drives Differential Immune Responses in GlioblastomaCancer DiscoverySeptember 6, 2023Juyeun Lee, Michael Nicosia, Ellen S. Hong, Daniel J. Silver, Cathy Li, Defne Bayik, Dionysios C. Watson, Adam Lauko, Kristen E. Kay, Sabrina Z. Wang, et al.Anti-TOX (TXRX10, cat. #12-6502) antibody was purchased from Invitrogen, and anti-TCF1 (C63D9, cat. #6709S) antibody was obtained from Cell Signaling Technology. Antibodies against PD-1 (RMP1-30, cat. #752354),

(5) SLC38A2 and glutamine signalling in cDC1s dictate anti-tumour immunityNatureAugust 1, 2023Guo C, You Z, Shi H, Sun Y, Du X, Palacios G, Guy C, Yuan S, Chapman NM, Lim SA, et al.Violet 605–anti-Ly108 (13G3, 745250, 1:200) (from BD Biosciences); Alexa Fluor 647–anti-TCF1 (C63D9, 6709, 1:100, Cell Signaling Technology). Migratory cDC1s and cDC2s in tumour dLN were respectively gated

(6) TCR-independent CD137 (4-1BB) signaling promotes CD8+-exhausted T cell proliferation and terminal differentiationImmunityJuly 1, 2023Andrea C. Pichler, Nadège Carrié, Marine Cuisinier, Samira Ghazali, Allison Voisin, Pierre-Paul Axisa, Marie Tosolini, Céline Mazzotti, Dominic P. Golec, Sabrina Maheo, et al.,Cell Signaling Technology,Cat#: 6444S Anti-human/Anti-Mouse TCF1/TCF7 PE [C63D9],Cell Signaling Technology,Cat#: 6709S Anti-mouse CD45.1-FITC (A20),BD Biosciences,Cat#: 110706RRID: AB_313495 Anti-mouse TCRb-Perc PCy5.5

(7) Intravenous heterologous prime-boost vaccination activates innate and adaptive immunity to promote tumor regressionCell ReportsJune 7, 2023Ramiro A. Ramirez-Valdez, Faezzah Baharom, Ahad Khalilnezhad, Sloane C. Fussell, Dalton J. Hermans, Alexander M. Schrager, Kennedy K.S. Tobin, Geoffrey M. Lynn, Shabnam Khalilnezhad, Florent Ginhoux, et al.Biosciences,Cat# 748291; RRID:AB_2872718 Anti-mouse TCF-1 (AF647); clone C63D9,Cell Signaling,Cat# 6709S; RRID:AB_2797631 Anti-mouse Tim-3 (BV605); clone RMT3-23,Biolegend,Cat# 119721; RRID:AB_2616907 Anti-mouse

(8) Lymphocyte networks are dynamic cellular communities in the immunoregulatory landscape of lung adenocarcinomaCancer CellMay 1, 2023Giorgio Gaglia, Megan L. Burger, Cecily C. Ritch, Danae Rammos, Yang Dai, Grace E. Crossland, Sara Z. Tavana, Simon Warchol, Alex M. Jaeger, Santiago Naranjo, et al.12–9668-82; RRID: AB_2572720 t-CyCIF: anti-human and mouse TCF1,Cell Signaling Technology,Clone C63D9; Cat# 6709; RRID: AB_2797631 t-CyCIF: anti-human and mouse Vimentin,Cell Signaling Technology,Clone D21H3; Cat#

(9) Multiparametric flow cytometry to characterize vaccine-induced polyfunctional T cell responses and T cell/NK cell exhaustion and memory phenotypes in mouse immuno-oncology modelsFrontiers in ImmunologyApril 6, 2023Davide Moi, Bijun Zeng, Simone A. Minnie, Rituparna Bhatt, Jack Wood, David P. Sester, Roberta Mazzieri, Riccardo Dolcetti2,NKp46,29A1.4,PE-Cy7,BL – 137618,1/200,Differentiation Markers specific for Panel 2,TCF-1*,C63D9,AF647,CST – 6709S,1/200,Stemness List of antibodies and relative fluorochromes used in Panels 1 & 2 . *Intracellular

(10) Visual Acuity in Patients Requiring Intravitreal Injections: Short-Term and Long-Term Effects of Delay in CareJournal of VitreoRetinal DiseasesApril 3, 2023Weilin Song, Emese Kanyo, Riley Bastian, Rishi P. Singh, Aleksandra V. Rachitskaya16,.0664 RVO,RVO,RVO,RVO,RVO,RVO Overall,319 ± 6,330 ± 7,.1066,296 ± 5,.0115 c Completed (n = 121),313 ± 10,308 ± 11,.6709,305 ± 9,.4377 Delayed (n = 85),329 ± 11,364 ± 13,.0467 c,282 ± 11,.0049 c Mean CST (µm) Between

(11) Extracellular acidosis restricts one-carbon metabolism and preserves T cell stemnessNature metabolismFebruary 1, 2023Cheng H, Qiu Y, Xu Y, Chen L, Ma K, Tao M, Frankiw L, Yin H, Xie E, Pan X, et al.25-5870-82, 1:200 for FACS) were purchased from Thermo Fisher. Alexa Fluor 647 anti-TCF1 (cat. no. 6709, 1:200 for FACS) and phospho-4E-BP1 (Thr37/46) (cat. no. 2846, 1:200 for FACS) were purchased from Cell Signaling Technology

(12) Tcf-1 protects anti-tumor TCR-engineered CD8+ T-cells from GzmB mediated self-destructionCancer Immunology, ImmunotherapyDecember 1, 2022Brendan Zangari, Takemasa Tsuji, Junko Matsuzaki, Hemn Mohammadpour, Cheryl Eppolito, Sebastiano Battaglia, Fumito Ito, Thinle Chodon, Richard Koya, A. J. Robert Robert McGray, et al.(BioLegend #424,401) and labelled intracellularly with αTcf-1 clone C63D9 (Cell Signaling Technologies #6709). LSR-II and Fortessa cytometers were used. In experiments detecting apoptosis, cells were labelled

(13) Impaired humoral immunity is associated with prolonged COVID-19 despite robust CD8 T cell responsesCancer CellJuly 11, 2022Olga Lyudovyk, Justin Y. Kim, David Qualls, Madeline A. Hwee, Ya-Hui Lin, Sawsan R. Boutemine, Yuval Elhanati, Alexander Solovyov, Melanie Douglas, Eunise Chen, et al.Biosciences,"751,572" CD138-BV605,BioLegend,"356,520" Eomes-PE-eFluor610,Invitrogen,61-4877-42 TCF-1-Alexa647,Cell Signaling,6709S CD38-BUV661,BD Biosciences,"612,969" CD95-BV650,BioLegend,"305,642" CCR7-BB700,BD Biosciences,"566,437" CD21-BV711,BD

(14) Functional assessment of the cell-autonomous role of NADase CD38 in regulating CD8(+) T cell exhaustioniScienceMay 20, 2022Ma K, Sun L, Shen M, Zhang X, Xiao Z, Wang J, Liu X, Jiang K, Xiao-Feng Qin F, Guo F, et al.AB_469504 PE-IL-2,Biolegend,Cat# 503808; RRID: AB_315302 Alexa Fluor647-anti-TCF-1,Cell Signaling Technology,Cat# 6709S; RRID: AB_2797631 PE-TOX,Invitrogen,Cat# 12-6502-82; RRID: AB_10855034 PerCP-Cy5.5-anti-Ki67,Biolegend,Cat#

(15) LSD1 inhibition sustains T cell invigoration with a durable response to PD-1 blockadeNature communicationsNovember 24, 2021Liu Y, Debo B, Li M, Shi Z, Sheng W, Shi Y.BioLegend, Cat#119703); Tim-3, APC (RMT3-23, BioLegend, Cat#119705); TCF1/TCF7, AF647 (C63D9, CST, Cat#6709S); CD127, PE-Cy7 (A7R34, BioLegend, Cat#135013); CD127, FITC (A7R34, BioLegend, Cat#135007); TOX, PE

(16) The CD155/TIGIT axis promotes and maintains immune evasion in neoantigen-expressing pancreatic cancerCancer cellOctober 11, 2021Freed-Pastor WA, Lambert LJ, Ely ZA, Pattada NB, Bhutkar A, Eng G, Mercer KL, Garcia AP, Lin L, Rideout WM 3rd, et al.PD-1,BD Biosciences,EH12.1 Cat# 563076 FC: anti-Human/Mouse TCF-1,Cell Signaling Technologies,C63D9 Cat# 6709S FC: anti-Human TIGIT,Invitrogen,MBSA43 Cat# 25–9500-41 FC: anti-Human TIM3,BD Biosciences,7D3 Cat#

(17) Conventional type I dendritic cells maintain a reservoir of proliferative tumor-antigen specific TCF-1+ CD8+ T cells in the tumor draining lymph nodesImmunityOctober 1, 2021Jason M. Schenkel, Rebecca H. Herbst, David Canner, Amy Li, Michelle Hillman, Sean-Luc Shanahan, Grace Gibbons, Olivia C. Smith, Jonathan Y. Kim, Peter M.K. Westcott, et al.(SAPE)",Invitrogen,Cat #: S21388 "Rabbit monoclonal anti TCF1/TCF7, Alexa Flour 647",Cell Signaling Technology,Cat #: 6709S; Clone: C63D9; RRID: AB_2797631; dilution: 1 to 250 "Rabbit monoclonal anti TCF1/TCF7, Alexa Flour

(18) Antigen dominance hierarchies shape TCF1(+) progenitor CD8 T cell phenotypes in tumorsCellSeptember 16, 2021Burger ML, Cruz AM, Crossland GE, Gaglia G, Ritch CC, Blatt SE, Bhutkar A, Canner D, Kienka T, Tavana SZ, et al.13G3; Cat# 742272; RRID: AB_2871448 FC: TCF1/TCF7,Cell Signaling Technology,"Clone C63D9; Cat# 6444, 6709, 9066; RRID: AB_2797627, AB_2797631, AB_2797696" FC: TIGIT,BD Biosciences,Clone 1G9; Cat# 565474; RRID:

(19) TCF-1 controls T_reg cell functions that regulate inflammation, CD8⁺ T cell cytotoxicity and severity of colon cancerNature immunologySeptember 1, 2021Osman A, Yan B, Li Y, Pavelko KD, Quandt J, Saadalla A, Singh MP, Kazemian M, Gounari F, Khazaie K.Q31-378; cat: 562607; BD Biosciences); and anti-TCF-1-Alexa Fluor 647 (dilution: 1/300; clone: C63D9; cat: 6709S; Cell Signaling) for 2 h or overnight at 4 °C. Cells were then washed twice with wash/perm buffer. For

(20) ST8Sia6 Promotes Tumor Growth in Mice by Inhibiting Immune ResponsesCancer Immunology ResearchAugust 1, 2021David J. Friedman, Sydney B. Crotts, Michael J. Shapiro, Matthew Rajcula, Shaylene McCue, Xin Liu, Khashayarsha Khazaie, Haidong Dong, Virginia Smith Shapiroanti-CD38-BV421 (Cat. #562768, BD Horizon) was purchased from BD Horizon, and anti-TCF1-AF647 (Cat. #6709S, Cell Signaling Technologies) was purchased from Cell Signaling Technologies. Fixable viability dye

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7/15/2020TCF1/TCF7 Rabbit mAb from CST Works Well on T Cels Puspa ThapaUsed it on mouse lung and draining lymph node T cells, after flu infection and it worked well. Was able to examine expression in naive (higher) and effector (lower) T cells.
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1/14/2019Good TCF1/TCF7 Antibody Rebecca GuyerOur lab studies the function of T cells in an influenza model in mice. One of our interests is to see how TCF1 expression may change due to different treatments we give the mice. This antibody is ... Read Review

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Cancer Immunology, Immunotherapy

Nature immunology

Nature immunology

Nature communications

Nature communications

TCF1/TCF7 (C63D9) Rabbit mAb (Alexa Fluor ® 647 Conjugate) from Cell Signaling Technology

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Cell Signaling Technology

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Webinar: Improving Cancer Biomarker Detection with Multiplex Technology

Screening of Leukemic Neoantigen Specific T cells with the iQue Screener PLUS

TCF1/TCF7 (C63D9) Rabbit mAb (Alexa Fluor ® 647 Conjugate) from Cell Signaling Technology

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