PA5-54578

antibody from Invitrogen Antibodies

Targeting: ZNF276 CENP-Z, CENPZ, MGC45417, ZADT, ZFP276, ZNF477

Antibody data

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Validation data

Product number: PA5-54578 - Provider product page
Provider: Invitrogen Antibodies
Product name: ZNF276 Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Recombinant full-length protein
Description: Immunogen sequence: PWDKETAPRL PQHRGWNPGD APQTSQGRGT GTPVGAETKT LPSTDVAQPP SDSDAVGPRS GFPPQPSLPL CRAPGQLGEK QLPSSTSDDR
Concentration: 0.1 mg/mL

ZNF276 protein structure - PA5-54578 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Expression and prognosis of ZNF276 in breast cancer tissues and cell lines. A mRNA expression of ZNF276 in different malignancies and normal tissues from TCGA database. BRCA breast carcinoma. CESC Cervical and endocervical carcinoma. COAD Colon adenocarcinoma. LIHC Liver hepatocellular carcinoma. LUAD Lung adenocarcinoma. STAD Stomach adenocarcinoma. UCEC Uterine Corpus Endometrial Carcinoma. B , C RT-qPCR ( B ) and western blot ( C ) analysis of ZNF276 expression in 10 breast cancer tissues and matched surrounding normal tissues. D Representative pictures of ZNF276 expression in 118 breast cancer tissues and 48 adjacent tissues from tissue chips. E , F Statistical analysis of ZNF276 protein expression in breast cancer tissues and adjacent tissues ( E ) and different molecular subtypes ( F ) from tissue chips by H-score. G ZNF276 mRNA expressions in different breast cancer cell lines were obtained from the CCLE portal. H Protein expression of ZNF276 was detected in normal breast cells and six breast cancer cells by western blot. I Correlation between ZNF276 expression and prognosis of breast cancer patients from TCGA dataset. J Kaplan-Meier curve showing the overall survival of 132 breast cancer patients separated by high and low ZNF276 expression. * p < 0.05; ** p < 0.01; *** p < 0.001.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 2 ZNF276 promotes the proliferation of breast cancer cells. A Western blot identifying the ZNF276 overexpression in MDA-MB-231 and SK-BR-3, ZNF276 knockdown in MCF-7 and ZNF276 knockout in UACC-812. B-C. CCK-8 assay measuring the effect of ZNF276 overexpression on cell proliferation in MDA-MB-231 ( B ) and SK-BR-3 ( C ) cells. D , E The effect of ZNF276 silencing ( D ) and knockout ( E ) on cell proliferation was evaluated by CCK-8 assay. F Cell proliferation ability of ZNF276 overexpressing cells was measured by clone formation assay. G Clone formation assay measuring the effect of ZNF276 silencing and knockout on cell proliferation. H ZNF276 overexpression affecting breast cancer tumor formation in the mammary fat pad of nude mice. I Measurement of tumor volume of the resected tumors from mice injected with ZNF276 overexpressing cells and control cells. J Statistical analysis of tumor weight of the resected tumors. K Immunohistochemical staining of Ki67 and ZNF276 in the resected tumors. Bar = 40 um. * p < 0.05; ** p < 0.01; *** p < 0.001.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 5 ZNF276 activates the Wnt/beta-catenin signaling by regulation of CYP1B1. A , C Western blotting ( A ) and RT-qPCR ( C ) analysis revealed that silencing of CYP1B1 inhibited the expression of c-Myc and Cyclin D1 enhanced by ZNF276 overexpression in MDA-MB-231 cells. B , D Effects of ZNF276 knockdown and CYP1B1 overexpression on c-Myc and Cyclin D1 expression were analyzed in UACC-812 cells by western blotting ( B ) and RT-qPCR ( D ). E , F ZNF276 lacking C 2 H 2 domain inhibited the TOP Flash activity enhanced by the intact ZNF276 with wnt3a activation in MDA-MB-231 ( E ) and UACC-812 ( F ) cells. G Endogenous beta-catenin expression and distribution was observed by immunofluorescence in MDA-MB-231 cells transfected with intact ZNF276 or ZNF276 lacking C 2 H 2 domain plasmid. Nuc nuclear. Bar = 20 um. * p < 0.05; ** p < 0.01; *** p < 0.001.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 7 ZNF276 activates Wnt/beta-catenin signaling through recruitment of MAGEB2 to regulate CYP1B1 transcription. A GO analysis of the potential ZNF276-interacted proteins. B KEGG analysis of the potential ZNF276-interacted proteins. C , D Both forward ( C ) and reverse ( D ) co-IP analysis affirmed the interaction between ZNF276 and MAGEB2. E ChIP-PCR analysis of ZNF276 binding ability at the CYP1B1 promoter affected by MAGEB2 overexpression. F , G The luciferase activity of CYP1B1 promoter decreased by ZNF276 silencing was further enhanced by the expression of MAGEB2 in 293T and MCF-7. H , I RT-qPCR ( H ) and western blot ( I ) analysis confirmed the effect of MAGEB2 overexpression in CYP1B1 mRNA and protein expression in MCF-7 cells. J , K The reduced transcriptional activity of CYP1B1 promoter by ZNF276 was partially reversed by MAGEB2. L , M Overexpression of MAGEB2 reversed the decreased cell proliferation, migration and invasion abilities affected by ZNF276 silencing in CCK-8 assay ( L ), transwell migration ( M , up) and transwell invasion ( M , down).