CD133 (D2V8Q) XP ® Rabbit mAb 64326S from Cell Signaling Technology

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Cell reports
July 06 2021

Fig 1: NELL2highCD133highEWS-FLI1high and NELL2lowCD133lowEWS-FLI1low populations in Ewing sarcoma display phenotypes consistent with high and low NELL2 signaling, respectively(A) NELL2 expression is heterogeneous in Ewing sarcoma tumor. NELL2 expression in surgically resected Ewing sarcoma tumor was examined by immunohistochemistry (brown signals). Scale bar: 60 µm.(B) NELL2 expression is heterogeneous in Ewing sarcoma cells and correlates with EWS-FLI1 expression. NELL2 and EWS-FLI1 expression in A-673 cells was assessed by anti-NELL2 (red) and anti-FLI1 C terminus (green) immunofluorescent staining. The nuclei were stained with DAPI. Scale bar: 10 µm.(C) NELL2 expression correlates with CD133 expression in Ewing sarcoma cells. NELL2 and CD133 expression in A-673 cells was assessed by anti-NELL2 (green) and anti-CD133 (AC133, red) immunofluorescent staining. The nuclei were stained with DAPI. Scale bar: 10 µm.(D) The CD133low population displays lower NELL2, EWS-FLI1, BRG1, BAF250A, and BAF155 levels than the CD133high population. A-673 cells were incubated with anti-CD133 (AC133) antibody and were sorted into the CD133high and CD133low populations. The expression of indicated proteins was assessed by immunoblotting.(E) The CD133low population displays reduced EWS-FLI1 target gene expression. The RNA expression of indicated genes was assessed by qRT-PCR and is presented after normalization to the levels in the CD133high population (blue). The expression of EWS-FLI1 target genes is reduced in the CD133low population (red) (n = 3).(F) The CD133low population displays slower growth than the CD133high population.(G) The CD133low population displays reduced NELL2 and EWS-FLI1 expression. A-673, CHLA-9, EW8, TC71, TC32, and SK-N-MC Ewing sarcoma cells were sorted into the CD133high and the CD133low populations, and the RNA expression of CD133, NELL2, and EWS-FLI1 was assessed by qRT-PCR (n = 3).(H) The CD133low population displays increased filopodia. Actin cytoskeleton was visualized by phalloidin staining (red). Cell nuclei were stained by DAPI (blue). Scale bars: 10 µm. The quantification of filopodia is shown in Figure S4D.(I) NELL2 silencing inhibits the proliferation of both CD133high and CD133low populations.(J) The CD133low population displays reduced sphere formation; *p < 0.05 (3 independent experiments).(K) The CD133low population displays reduced xenograft tumorigenicity (n = 5, p < 0.05).(L) The CD133low population displays reduced migration; *p < 0.05 (3 independent experiments).(M) Prolonged cisplatin or doxorubicin treatment enriches the CD133low population.
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Cancer communications (London, England)
October 01 2022

Fig 2: HK2 interacts with CD133 and inhibits polyubiquitylation-dependent CD133 degradation. (A-B) HK2 was depleted (A) or overexpressed (B) in H1048 or H69 cells. Immunoblotting analyses were performed with an anti-CD133 antibody. (C-D) H1048 cells with or without HK2 depletion (C) or overexpression (D) were treated with CHX (100 µg/mL) for the indicated periods of time. Immunoblotting analyses were performed with the indicated antibodies (left). Quantification of CD133 expression levels relative to tubulin expression levels is shown (right). (E-F) H1048 cells with or without HK2 depletion (E) or overexpression (F) were treated with MG132 (50 µmol/L) for 8 hours. Immunoprecipitation with an anti-CD133 antibody and immunoblotting analyses were performed with the indicated antibodies. (G) Purified His-HK2 was incubated with purified GST or GST-CD133 together with glutathione-sepharose 4B beads. GST pulldown assay and immunoblotting analyses were performed with the indicated antibodies. (H) Mitochondrial and non-mitochondrial fractions of H1048 monolayers or spheroids were prepared and subjected to immunoprecipitation with an anti-HK2 antibody. Immunoblotting analyses were performed with the indicated antibodies. (I) RNAi-resistant WT HK2, inactive HK2 D209/657A, or HK2 ?N16 was expressed in H1048 and H69 cells with depletion of endogenous HK2. Immunoblotting analyses were performed with the indicated antibodies. (J) H1048 cells with or without WT HK2 or inactive HK2 D209/657A overexpression were treated with MG132 (50 µmol/L) for 8 hours. Immunoprecipitation and immunoblotting analyses were performed with the indicated antibodies. (K) H1048 cells with or without expression of inactive HK2 D209/657A were treated with CHX (100 µg/mL) for the indicated periods of time. Immunoblotting analyses were performed with the indicated antibodies (upper). Quantification of CD133 expression levels relative to tubulin expression levels is shown (lower). Abbreviations: HK2, hexokinase 2; CHX, cycloheximide; IP, immunoprecipitation; WCL, whole cell lysate; WT, wide type
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Annals of Translational Medicine
June 01 2021

Fig 3: SNTB1 silencing reduces expression of stemness markers, inhibits spherogenicity, and decreases SP. (A) Heatmap plot of differentially expressed cancer stem cell-related genes in shSNTB1 versus shVec. cells. Upregulated genes: depicted in red, log2 fold change >2; downregulated genes: depicted in blue, log2 fold change <–2. (B) Detection of the cancer stem cell surface markers, CD133, LGR5, and EpCAM using Western blot. Quantification of cancer stem cell surface protein levels are normalized to the α-tubulin level. (C) Representative micrographs of tumor spheres formed by the indicated SW620 and CW-2 cells with shVec. or shSNTB1 transduction (200×). Histograms show the percentage of sphere formation. (D) Flow cytometry-based Hoechst 33342 efflux assay evaluates the proportion of the SP in shSNTB1 versus shVec. cells. Histograms show the percentage of SP cells. *, P<0.05. SNTB1, beta-1 syntrophin; SP, side population; LGR5, leucine-rich repeat-containing G-protein-coupled receptor 5; EpCAM, epithelial cell adhesion molecule.
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Frontiers in oncology
January 01 2022

Fig 4: The miR-4800-3p promotes the expression of tumor stem cell markers and EMT in low-metastatic HCC cells through regulating the Hippo pathway. (A) The expression of miR-4800-3p was measured by qPCR assay in low-metastatic HepB3 and LM3 cells treated as previously described. STK25 expression was detected by qPCR (B), Western blot analysis (C), and immunofluorescence assay (D), respectively. qPCR and Western blot analysis of tumor stem cell markers (CD44, CD133, and OCT4) (E, F) and EMT biomarkers (E-cadherin, ZO-1, N-cadherin) (G, H) in low-metastatic HepB3 and LM3 cells. (I) Western blot was used to detect the expression of p-YAP, YAP, TAZ, and PCNA in HepB3 and LM3 cells. *p < 0.05, **p < 0.01 vs. control. ## p < 0.01 vs. NC.
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International Journal of Stem Cells
August 30 2021

Fig 5: Combination use of BJJP and BMSCs further suppress the cell growth of CSCs in addition to enhancing cell apoptosis. (A, B) the expressions of CSCs biomarkers CD24, CD133 and EpCAM were measured with RT-qPCR and western blot; (C) cell proliferation ability was assessed by cell clone formation assay; (D) cell viability was determined by CCK-8 assay; (E) cell apoptosis rate was assessed by Flow cytometry. N=3, *p<0.05, **p<0.01; EpCAM, epithelial cell adhesion molecule; CSCs, cancer stem cells; BMSCs, bone mesenchymal stem cells; BJJP, Bie Jia Jian pill.

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CD133 (D2V8Q) XP ® Rabbit mAb

Cell Signaling Technology's CD133 (D2V8Q) XP ® Rabbit mAb is a Rabbit monoclonal antibody. The CD133 (D2V8Q) XP ® Rabbit mAb Antibody was generated using PROM1 as the antigen. It reacts with Homo Sapiens, and Human. This antibody has been shown to work in applications such as: Immunocytochemistry, Immunofluorescence, Immunohistochemistry, Immunohistochemistry - fixed, and Western Blot.

Description

CD133 (D2V8Q) XP® Rabbit mAb recognizes endogenous levels of total CD133 protein

Product Specs
ItemCD133 (D2V8Q) XP ® Rabbit mAb
CompanyCell Signaling Technology
Price $402.00 Inquire
Catalog Number64326S
Quantity100 µl
ApplicationsW, IHC-Bond, IHC-P, IF-IC
ReactivityH
TargetCD133
HostRabbit
NCBI Full Gene Nameprominin 1
NCBI Gene AliasesAC133, CD133, CORD12, MCDR2, MSTP061, PROML1, RP41, STGD4
ClonalityMonoclonal
Research AreaDevelopmental Biology
Molecular Weight133

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Questions and Answers

Has this antibody been used in studies related to cancer stem cells?

Yes, the CD133 (D2V8Q) XP® Rabbit mAb has been utilized in various studies focusing on cancer stem cells. For instance, it was employed in research demonstrating the role of FTO in maintaining cancer stem cell properties in pancreatic cancer, where depletion of FTO led to reduced expression of cancer stem cell markers, including CD133 ().

Can this antibody be used for immunofluorescence applications?

Yes, the CD133 antibody has been shown to work effectively in immunofluorescence applications. It was used in studies to assess the expression of CD133 in three-dimensional cell cultures, highlighting its utility in visualizing stem cell markers in various cancer models ().

What is the significance of CD133 in cancer research?

CD133 is recognized as a marker for cancer stem cells and is implicated in tumorigenesis and metastasis. Studies have shown that targeting CD133 can affect the self-renewal and differentiation of cancer stem cells, making it a critical focus in understanding cancer biology and developing therapeutic strategies (; ).

Is there evidence supporting the use of this antibody in Western blotting?

Yes, the CD133 antibody has been validated for use in Western blotting. It has been cited in multiple studies where it was used to detect CD133 protein levels in various cancer cell lines, contributing to the understanding of stemness and tumor progression (; ).

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(1) Stemness and Survival: CD117+/CD133+ Subpopulations Sustain PI3K Signaling and Drive Imatinib Resistance in Head and Neck Mucosal MelanomaCellsApril 19, 2026Sofie-Yasmin Hassan, Simeon Santourlidis, Thomas W. Flanagan, Sarah-Lilly Hassan, He Zhou, Morna F. Schmidt, Claudio Cacchi, Matthias Ferdinand Lammert, Mossad Megahed, Amir Sadegh Yazdi, et al.histochemistry, including anti-CD117/c-Kit [(D13A2) XP® Rabbit mAb #3074], CD133 [(D2V8Q) XP® Rabbit mAb #64326], anti-CD133 (#64326), c-Kit (#37805), p85 (#4257), and Actin (#8457), and Phospho-PDK1 [Ser241] (#3061)

(2) Dual-targeting CD133/PD-L1 CAR-T plus αPD-1 overcomes immunosuppressive microenvironment and enhanced by radiation pre-conditioningMolecular TherapyMarch 4, 2026Zhuoran Yao, Kai Kang, Pei-Heng Li, Limei Yin, Ruizhan Tong, Linglu Yi, Yonghong Song, Ren Luo, Yijun Wu, Shanghai Liu, et al.performed. The sections were incubated overnight at 4°C with an anti-CD133 monoclonal antibody (1:50) (64326s, Cell Signaling Technology) followed by an incubation with a secondary antibody (goat anti-rabbit/horseradish

(3) A human patient-derived organoid biobank to model tumor heterogeneity and therapeutic vulnerability for oral squamous cell carcinomaCell Reports MedicineFebruary 17, 2026Yuhan Wang, Pengfei Diao, Pin Lv, Na Qi, Ziyu Wang, Enshi Yan, Jin Li, Yanling Wang, Dongmiao Wang, Yaping Wu, et al.AB_2061561 Anti-CD44,Cell Signaling Technology,Cat#3570; RRID: AB_2076465 Anti-CD133,Cell Signaling Technology,Cat#64326; RRID: AB_2721172 Anti-BMI1,Cell Signaling Technology,Cat#6964; RRID: AB_10828713 Anti-β-catenin,Proteintech,Cat#51067-2-AP;

(4) Programmable iPSC-derived CAR-NK vesicles remodel the immune microenvironment and eradicate tumorsCell Reports MedicineFebruary 17, 2026Hao Zhang, Shenglong Li, Chongzhong Liu, Xiangdong Gongye, Han Li, Yiyi Ji, Cheng-Wei Ju, Wenzhen Jia, Xing Niu, Yujing Guan, et al.serum for 30 min at RT. Sections were incubated overnight at 4°C with anti-CD133 antibody (CST, Cat#64326; 1:200 dilution) diluted in PBS containing 1% BSA. Slides were washed and incubated with HRP-linked

(5) Potent synthetic lethality between PLK1 and EYA family inhibitors in tumors of the central and peripheral nervous systemsGenes & DevelopmentFebruary 2, 2026Christopher B. Nelson, Jadon K. Wells, Ekaagra Kesarwani, Alexander P. Sobinoff, Madushan Fernando, Jixuan Gao, Karen L. MacKenzie, Rebecca C. Poulos, Urwah Nawaz, Xiang Wang, et al.V9131), p-S10 H3 (1:1000; Cell Signaling Technology 9706), CD133 (1:1000; Cell Signaling Technology 64326), and Sox9 (1:1000; Cell Signaling Technology 82630). Densitometry was performed in ImageJ on unaltered

(6) DCLK1 drives malignant progression and chemoresistance of bladder cancer by deubiquitinating HDAC6Molecular CancerJanuary 30, 2026Ashuai Du, Yuzheng Zhou, Xinpei Deng, Dongbo Yuan, Kai Li, Yuanyuan Luo, Songsong Tan, Xuchao Dai, Bo Yu, Kehua Jiang, et al.(1:1000; Proteintech, 66031-1-Ig), ALDH1 (1:1000; Cell Signaling, 54135), CD133 (1:1000; Cell Signaling, 64326), CD44 (1:1000; Cell Signaling, 37259),SOX2 (1:1000; Cell Signaling, 23064),Nanog (1:2000; Cell Signaling,

(7) (Non)canonical Wnt signaling, cytoarchitecture and stemness: new insights from primary nonmetastatic, primary metastatic, regional and distant metastatic models of adrenocortical carcinomaExperimental & Molecular MedicineNovember 28, 2025Igor Shapiro, Huguette Debaix, Chiara Kräuchi, Andrea Abate, Stefan R. Bornstein, Svenja Nölting, Samuel Gunz, Alfredo Berruti, Sandra Sigala, Felix Beuschlein, et al.USA)",1/800 Nanog,"4903, Cell Signaling (Danvers, MA, USA)",1/200 CD68,"ab955, Abcam (Cambridge, UK)",1/100 CD133,"64326, Cell Signaling (Danvers, MA, USA)",1/100 ALDH1A1,"ab52492, Abcam (Cambridge, UK)",1/200 CD24,"67627-1,

(8) Amlodipine exerts inhibitory effects against glioma stem cells through degrading EGFR and down-regulating its downstream pro-survival pathwaysCell Death DiscovOctober 27, 2025Zengyang Li, Xiaopei Zhang, Ping Wen, Fan Ni, Nanheng Yin, Zhicheng Zhang, Tao Zhong, Feiyu Xia, Jiaxin Pan, Liang Liu, et al.Akt (CST, #9272), p-Akt (Ser473) (CST, #9271), mTOR (CST, #2983), p-mTOR (CST, #2971), CD133 (CST, #64326), SOX2 (CST, #3579), EGFR (CST, #4267), p-EGFR (CST, #3777), Ubiquitin (CST, #20326), Na/K-ATPase (CST,

(9) Endothelial cell-derived SDF-1α elicits stemness traits of glioblastoma via dual-regulation of GLI1TheranosticsSeptember 8, 2025Ye Yuan, Xudong Liu, Liwen Kuang, Shixue Yang, Lihong Wang, Jiao Wang, Sen Wei, Zexuan Yan, Qinghua Ma, Juan Lei, et al.antibodies overnight at 4 °C after dilution in 5% BSA. The primary antibodies used were CD133 (1:50, CST, #64326, USA) and αSMA (1:100, Abcam, ab7817, USA). After three washes, the cells were stained with secondary

(10) Targeting CAFs-Mediated Stromal Signaling in a Patient-Derived Organotypic Colorectal Tumor ModelMolecular cancer therapeuticsAugust 1, 2025Lamichhane A, Guragain P, Heiss J, Rafsanjani Nejad P, Rana Magar A, Ciavattone N, Agrawal S, Luker GD, Tavana H.AB_2143884), MET (Cat. No 8198, RRID: AB_10858224), GAPDH (Cat. No 5174, RRID: AB_10622025), CD133 (Cat. No 64326, RRID: AB_2721172), phospho-Rb (Ser807/811, Cat. No 8516, RRID: AB_11178658), Rb (Cat. No 9313, RRID:

(11) USP18 deubiquitinates and stabilizes SOX9 to promote the stemness and malignant progression of glioblastomaCell Death DiscovMay 15, 2025Zhiyuan Liu, Kuo Yu, Kaile Chen, Yi Zhang, Kexiang Dai, Liang Zhao, Peng Zhaoantibodies used for WB analysis were purchased from Cell Signaling Technology (#4813, #5568, #3580, #64326, #3579, #73349, #82630, #2276, #3724, #2624, #46395, #2743) or Abcam (ab205606). Following incubation

(12) Targeting metabolic vulnerability by combining NAMPT inhibitors and disulfiram for treatment of recurrent ovarian cancerCell Death DisApril 25, 2025Kei Kudo, Yoshimi Endo Greer, Daniel R. Crooks, Ye Yang, Jeffrey R. Brender, Teruhiko Yoshida, Brittney S. Harrington, Rahul Kamdar, Soumya Korrapati, Yusuke Shibuya, et al.USA) and used at the indicated dilutions for western blot analysis: CD44 (#3570S, 1:1000), CD133 (#64326S, 1:1000), PBEF/NAMPT (#86634S, 1:1000), lamin B1 (#12586S, 1:1 000), hexokinase II (#2867S, 1:1000),

(13) 7‑Difluoromethoxyl‑5,4'‑di‑n‑octylygenistein targets the STAT3 pathway by upregulating microRNA‑152‑3p expression to inhibit self‑renewal and tumor growth in non‑small cell lung carcinomaOncology ReportsApril 16, 2025Qing Yuan, Xiang Li, Xuemei Chen, Jianhui Xiao, Jiansong Zhang1:2,000 dilution; cat. no. 9145; Cell Signaling Technology, Inc.), CD133 (1:1,000 dilution; cat. no. 64326; Cell Signaling Technology, Inc.), CD44 (1:1,000 dilution; cat. no. 37259; Cell Signaling Technology,

(14) Histone lactylation enhances GCLC expression and thus promotes chemoresistance of colorectal cancer stem cells through inhibiting ferroptosisCell Death DisMarch 20, 2025Jiao Deng, Yangkun Li, Lanlan Yin, Shuang Liu, Yanqi Li, Wancheng Liao, Lei Mu, Xuelai Luo, Jichao Qindescribed. Antibodies and chemicals The following commercially available primary antibodies were used: Anti-CD133 (64326; Cell Signaling Technology); Anti-Nanog (8800; Cell Signaling Technology); Anti-OCT4A (83932; Cell Signaling Technology

(15) Comparative analysis of pediatric SHH medulloblastoma DAOY spheres and adherent monolayers: implications for medulloblastoma researchCancer Cell IntJanuary 22, 2025Jana Klementová, Šárka Jarošová, Irina Danilová, Markéta Farníková, Josef Novotný, Marie Davídková, Martina Zíková(TBST). Filters were incubated in primary antibody (rabbit polyclonal CD133 antibody, D2V8Q, 1:800, 64326 Cell Signaling; rabbit polyclonal actin antibody, 1:1000, A2066, Sigma-Aldrich; mouse monoclonal MAP2

(16) MAPK14/p38α shapes the molecular landscape of endometrial cancer and promotes tumorigenic characteristicsCell ReportsJanuary 1, 2025Sayali Joseph, Xingyuan Zhang, Gaith N. Droby, Di Wu, Victoria Bae-Jump, Scott Lyons, Angie Mordant, Allie Mills, Laura Herring, Blake Rushing, et al.(Ser139),Millipore Sigma,Cat#05–636; RRID:AB_309864 Rabbit monoclonal anti-CD133,Cell signaling technology,Cat#64326; RRID:AB_2721172 Rabbit monoclonal anti-SOX2,Cell signaling technology,Cat#3579; RRID:AB_2195767 Mouse

(17) Natural product mediated mesenchymal–epithelial remodeling by covalently binding ENO1 to degrade m6A modified β-catenin mRNAActa Pharmaceutica Sinica BJanuary 1, 2025Tianyang Chen, Guangju Liu, Sisi Chen, Fengyuan Zhang, Shuoqian Ma, Yongping Bai, Quan Zhang, Yahui Dinganti-E-cadherin (Cell Signaling Technology, 14472, dilution 1:1000), anti-CD133 (Cell Signaling Technology, 64326, dilution 1:1000), anti-SOX2 (Cell Signaling Technology, 3579, dilution 1:1000), anti-β-catenin (Cell Signaling Technology

(18) FDFT1 maintains glioblastoma stem cells through activation of the Akt pathwayStem Cell Research & TherapyDecember 20, 2024Hui Mo, Jiajia Shao, Zhun Li, Peiting Zeng, Xinke Yin, Yongsheng Huang, Peng Wang, Jianwei Liaoimmunodecorated with antibodies specific for FDFT1 (abcam, #ab236666), CD133 (Cell Signaling Technology, CST, #64326S), GFAP (CST, #80788S), SREBP1 (Abclonal, #A15586), SREBP2 (Abclonal, #A13049), PI3K (Abclonal, # A22730),

(19) Prion protein regulates invasiveness in glioblastoma stem cellsBMC CancerDecember 18, 2024Mariana B. Prado, Bárbara P. Coelho, Rebeca P. Iglesia, Rodrigo N. Alves, Jacqueline M. Boccacino, Camila F. L. Fernandes, Maria Isabel Melo-Escobar, Shamini Ayyadhury, Mario C. Cruz, Tiago G. Santos, et al.(ab703, Abcam), anti-CD44 (#3570, Cell Signaling Technology), anti-CD133 (Cell Signalig Technology, #64326), anti-SSEA1 (Cell Signaling Technology, #4744S) 1:50 in 0.5% BSA in PBS for 30 min on ice, following

(20) Isolation and identification of patient-derived liver cancer stem cells and development of personalized treatment strategiesJournal of translational medicineNovember 18, 2024Guo T, Zhang S, Zeng W, Liang Y, Xie J, Liu S, Qiu Y, Fu Y, Ou Y, Ma K, et al.°C overnight targeting AFP (MAB1368, R&D system), CD44 (3570 S, Cell Signaling Technology), CD133 (64326 S, Cell Signaling Technology), CD31 (AF23628-SP, R&D system), ALB (ab106582, abcam), NCAM (3576 S, Cell Signaling Technology

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Cell reports

Cancer communications (London, England)

Annals of Translational Medicine

Frontiers in oncology

International Journal of Stem Cells

CD133 (D2V8Q) XP ® Rabbit mAb from Cell Signaling Technology

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Cell Signaling Technology

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CD133 (D2V8Q) XP ® Rabbit mAb from Cell Signaling Technology

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