Invitrogen

HOXB3 Polyclonal Antibody

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HOXB3 Antibody in Immunocytochemistry (ICC/IF)

HOXB3 Antibody in Immunocytochemistry (ICC/IF)

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HOXB3 Antibody (PA5-103890) in ICC/IF

Immunocytochemistry analysis of HOXB3 in A549 cells. The samples were fixed with PFA and permeabilized in 0.1% Triton X-100, then blocked in 10% serum for 45 minutes at 25°C. Samples were incubated with HOXB3 polyclonal antibody (Product # PA5-103890) and mouse anti-beta tubulin antibody for 1 hour at 37°C. An AlexaFluor594 conjugated goat anti-rabbit IgG(H+L) (Red) and an AlexaFluor488 conjugated goat anti-mouse IgG(H+L) (Green) were used as the secondary antibodies. The nuclear counter stain is DAPI (blue).

HOXB3 Antibody in Immunocytochemistry (ICC/IF)

HOXB3 Antibody in Immunohistochemistry (Paraffin) (IHC (P))

HOXB3 Antibody in Immunohistochemistry (Paraffin) (IHC (P))

HOXB3 Antibody in Western Blot (WB)

HOXB3 Antibody in Western Blot, Immunocytochemistry (WB, ICC/IF)

HOXB3 Antibody in Western Blot, Immunoprecipitation (WB, IP)

HOXB3 Antibody in Western Blot (WB)

HOXB3 Antibody in Western Blot (WB)

HOXB3 Antibody in ChIP Assay (ChIP)

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HOXB3 Antibody in Immunocytochemistry (ICC/IF) — Immunocytochemistry analysis of HOXB3 in A549 cells. The samples were fixed with PFA and permeabilized in 0.1% Triton X-100, then blocked in 10% serum for 45 minutes at 25°C. Samples were incubated with HOXB3 polyclonal antibody (Product # PA5-103890) and mouse anti-beta tubulin antibody for 1 hour at 37°C. An AlexaFluor594 conjugated goat anti-rabbit IgG(H+L) (Red) and an AlexaFluor488 conjugated goat anti-mouse IgG(H+L) (Green) were used as the secondary antibodies. The nuclear counter stain is DAPI (blue).

HOXB3 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of HOXB3 in mouse brain tissue. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. Samples were incubated with HOXB3 polyclonal antibody (Product # PA5-103890) using a dilution of 1:100 (4°C overnight) followed by HRP conjugated anti-Rabbit secondary antibody.

HOXB3 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of HOXB3 in rat brain tissue. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. Samples were incubated with HOXB3 polyclonal antibody (Product # PA5-103890) using a dilution of 1:100 (4°C overnight) followed by HRP conjugated anti-Rabbit secondary antibody.

HOXB3 Antibody in Western Blot (WB) — Western blot analysis of HOXB3 in hybridoma cell lysate (Lane 1: treated with blocking peptide). Samples were incubated with HOXB3 polyclonal antibody (Product # PA5-103890).

HOXB3 Antibody in Western Blot, Immunocytochemistry (WB, ICC/IF) — Fig. 4 HOXB3 can be activated by extra WNT3A and the dysfunction of destruction complex, see also Fig. S2 . A HEK293 cells were transfected with CDCA3-Luc and siRNAs for control (siCtrl) or HOXB3 (siHOXB3). Transfected cells were cultured in medium with or without WNT3A and IWR-1 (enhancing AXIN1 stabilization). The results of western blots (for HOXB3, beta-catenin and beta-actin), luciferase assay (for CDCA3-Luc recording HOXB3-dependent transcriptional activity), and qRT-PCR assay (for HOXB3) were presented. Data were normalized to control (lane 4). B HEK293 cells were transfected with CDCA3-Luc and siRNAs (siCtrl, siHOXB3, or sibeta-catenin). Transfected cells were cultured in medium with or without WNT3A. The results of western blots (beta-catenin and beta-actin) and luciferase assay (CDCA3-Luc) of the indicated samples were presented. In luciferase assay, data were normalized to control (lane 1). C Top: Western blot of HOXB3 in HEK293 cells as indicated, transfected with siRNAs targeting control (siCtrl), AXIN1 (siAXIN1), APC (siAPC), or GSK3 (siGSK3), followed by another siRNAs targeting control or HOXB3. Bottom: luciferase assays of TOP-FLASH (beta-catenin/TCF-dependent transcriptional activity) or CDCA3-Luc reporters in the same cells used for western-blot. D Western blots of HOXB3, AXIN1, APC and beta-actin in LNCaP-AI cells transfected with siRNAs targeting control (siCtrl), AXIN1 (siAXIN1) or APC (siAPC). E qRT-PCR assay of CDCA3 in LNCaP-AI cells after stable tran

HOXB3 Antibody in Western Blot, Immunoprecipitation (WB, IP) — Fig. 5 HOXB3 can be degraded by beta-TrCP in destruction complex depend on the kinase activity of CKIalpha, see also Fig. S3 . A Western blot of indicated proteins CO-IP with HOXB3 in LNCaP-AI cells cultured in medium with or without WNT3A. B Graph showing HOXB3 structure with distinguishable functional domains distinguished []: HTLV-II domain (N-terminal), Homeobox domain, and DUF4074 domain (C-terminal). C Autoradiography of 35S-AXIN1 pulled by GST-fused full-length or truncated HOXB3 as indicated. Input: western blot of GST fused truncated HOXB3 the same as samples used in pull-down experiments. D Western blot of beta-TrCP and HOXB3 in LNCaP-AI cells treated with siRNAs targeting control (siCtrl) or beta-TrCP (sibeta-TrCP). E Western blot of AXIN1 and beta-TrCP CO-IP with HOXB3 in LNCaP-AI cells after deletion of HOXB3, IKKbeta, CKIalpha, or GSK3. Inputs: Western blot of HOXB3, AXIN1 and beta-TrCP in the same lysates. F Western blot of AXIN1, beta-TrCP and HOXB3 CO-IP with CKIalpha in WNT3A-treated LNCaP-AI cells that harvested at different time points as indicated. Inputs: Western blot of CKIalpha, HOXB3, AXIN1 and beta-TrCP in the same lysates as above. G Graph showing phospho-degrons in HOXB3 and other proteins that may be recognized by CKIalpha, IKKbeta, or GSK3. Diagrammatic sketch in frame showing three induced mutants of HOXB3 (M1, M2, and M3 for short) and mutation sites. H Top: western blot of HOXB3, CKIalpha and beta-actin in LNCaP-AI cells transfected with siCtr

HOXB3 Antibody in Western Blot (WB) — Fig. 3 HOXB3 protein, but not mRNA, correlates with WNT3A Expression in PCA. A Heatmap showing differentially expressed genes in aPCA with negative (HOXB3-, n = 10) and high score (HOXB3+, n = 10) of HOXB3 determined by RNA-Seq (see also Table S2 ). B Volume plot showing the significantly different expression of WNT3A between HOXB3+ and HOXB3- tumors determined by RNA-Seq (see also Table S2 ). C Scatter plots with bar showing individual expression of WNT3A, HOXB3, and CDCA3 (target gene of HOXB3) in both HOXB3- and HOXB3 + groups determined by RNA-Seq (see also Table S2 ). D qRT-PCR measured WNT3A, HOXB3, and CDCA3 mRNA expression in both HOXB3- and HOXB3 + groups ( n = 10 for each group). E Western blot of WNT3A, HOXB3, and CDCA3 protein expression in both HOXB3- and HOXB3 + groups ( n = 10 for each group). Error bars indicate mean +- SD. F Quantified results of the WNT3A and HOXB3 protein expression by Image J. G The significant correlation between WNT3A and HOXB3 protein expression in Fig. 3E should be determined by Pearson Correlation Coefficient.

HOXB3 Antibody in Western Blot (WB) — Figure 5 HOXB3 is found to be a downstream target of miR-375. (a) Binding sites between miR-375 and HOXB3 predicted through an online website. (b) The target relationship between miR-375 and HOXB3 was confirmed by dual-luciferase reporter gene assay. (c) HOXB3 mRNA expression in SNU-449 and Huh7 cells using RT-qPCR. (d, e) HOXB3 protein expression in SNU-449 and Huh7 cells using Western blot assay. (f) Enrichment analysis of miR-375 targeted genes using KEGG enrichment analysis. * p < 0.05 vs. the NC mimic group; # p < 0.05 vs. the EV-NC mimic group. All data were expressed as mean +- standard deviation of at least three independent experiments. Data analysis among multiple groups was performed by two-way ANOVA, followed by Tukey's multiple comparison test.

HOXB3 Antibody in ChIP Assay (ChIP) — Fig. 6 HOXB3 transcriptionally drives multiple WNT-regulated genes and is required for abiraterone resistance in APC-deficient CRPC cells, see also Fig. S4 . A GSEA of the APC-KO associated genes in data set of GSE144325 comparing differentially expressed genes in HOXB3 high HOXB3 negative tumors. Normalized enrichment score (NES) = 1.87, p = 0.0045. B Venn diagram showing genes significantly up-regulated in HOXB3 high and APC-KO samples. Overlapped genes showing significant enrichments in cell proliferation determined by Functional Annotation Tool [], including RAS, PI3K-AKT and MAPK signals (See also Tables S3 , S4 ). C LNCaP-AI cells were stably infected by shScramble (short for LA-shScr), shAPC (short for LA-shAPC), or by shAPC and shHOXB3 (short for LA-APC/HOXB3), followed by qRT-PCR assay for the overlapped genes in these cells. Heat maps showing the results of PCR that shAPC upregulated almost all the overlapped genes, while most of them overturned by additional HOXB3 knockdown. D qRT-PCRs assay based on CHIP experiments showing HOXB3 binds to the promoters of representative WNT-regulated genes. ChIP experiments using anti-HOXB3 antibody or normal IgG were performed in LA-shAPC cells. E The pGL3-CDCA3 was co-transfected with pcDNA3.1-HOXB3 or pcDNA3.1-vector into LA-shScr, LA-shAPC, and LA-APC/HOXB3 cells. Luciferase assays for promoter activities of representative WNT-regulated genes in indicated cells 24 h post transfection. F , G Western-blot and Bar graph showing u

产品信息

PA5-103890

应用

建议稀释比

已发表文章

免疫印迹 (WB)

1:1,000-1:3,000

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免疫组化（石蜡） (IHC (P))

1:50-1:200

-

免疫细胞化学 (ICC/IF)

1:100-1:500

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免疫沉淀 (IP)

-

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染色质免疫沉淀实验 (ChIP)

-

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产品规格

种属反应

Human, Mouse, Rat

已发表种属

Not Applicable

宿主/亚型

Rabbit / IgG

分类

Polyclonal

类型

Antibody

抗原

A synthesized peptide derived from human HOXB3(Accession P14651), corresponding to amino acid residues A359-D409.

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偶联物

Unconjugated Unconjugated Unconjugated

形式

Liquid

浓度

1 mg/mL

纯化类型

Affinity chromatography

保存液

PBS, pH 7.4, with 50% glycerol

内含物

0.02% sodium azide

保存条件

-20°C

运输条件

Wet ice

AB_2853222

产品详细信息

Antibody detects endogenous levels of total HOXB3.

靶标信息

Sequence-specific transcription factor which is part of a developmental regulatory system that provides cells with specific positional identities on the anterior-posterior axis.

仅用于科研。不用于诊断过程。未经明确授权不得转售。

Product References

HOXB3 drives WNT-activation associated progression in castration-resistant prostate cancer.

Cell death & disease

Zhu S,Yang Z,Zhang Z,Zhang H,Li S,Wu T,Chen X,Guo J,Wang A,Tian H,Yu J,Zhang C,Su L,Shang Z,Quan C,Niu Y

Published figure using HOXB3 polyclonal antibody (Product # PA5-103890) in ChIP Assay

Mon Mar 27 00:00:00 UTC 2023

HOXB3 drives WNT-activation associated progression in castration-resistant prostate cancer.

Cell death & disease

Zhu S,Yang Z,Zhang Z,Zhang H,Li S,Wu T,Chen X,Guo J,Wang A,Tian H,Yu J,Zhang C,Su L,Shang Z,Quan C,Niu Y

Published figure using HOXB3 polyclonal antibody (Product # PA5-103890) in ChIP Assay

Mon Mar 27 00:00:00 UTC 2023

HOXB3 drives WNT-activation associated progression in castration-resistant prostate cancer.

Cell death & disease

Zhu S,Yang Z,Zhang Z,Zhang H,Li S,Wu T,Chen X,Guo J,Wang A,Tian H,Yu J,Zhang C,Su L,Shang Z,Quan C,Niu Y

Published figure using HOXB3 polyclonal antibody (Product # PA5-103890) in ChIP Assay

Mon Mar 27 00:00:00 UTC 2023

HOXB3 drives WNT-activation associated progression in castration-resistant prostate cancer.

Cell death & disease

Zhu S,Yang Z,Zhang Z,Zhang H,Li S,Wu T,Chen X,Guo J,Wang A,Tian H,Yu J,Zhang C,Su L,Shang Z,Quan C,Niu Y

Published figure using HOXB3 polyclonal antibody (Product # PA5-103890) in ChIP Assay

Mon Mar 27 00:00:00 UTC 2023

Extracellular Vesicle-Encapsulated MicroRNA-375 from Bone Marrow-Derived Mesenchymal Stem Cells Inhibits Hepatocellular Carcinoma Progression through Regulating HOXB3-Mediated Wnt/β-Catenin Pathway.

Analytical cellular pathology (Amsterdam)

Yu Z,Liu J,Fan Q,Yu J,Ren X,Wang X

Published figure using HOXB3 polyclonal antibody (Product # PA5-103890) in Western Blot

Mon Apr 04 00:00:00 UTC 2022

生物信息学

蛋白别名: homeo box 2G; homeo box B3; Homeobox protein Hox-2.7; Homeobox protein Hox-2G; Homeobox protein Hox-B3; Homeobox protein MH-23

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基因别名: Hox-2.7; HOX2; HOX2G; Hoxb-3; HOXB3

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UniProt ID: (Human) P14651, (Mouse) P09026

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Entrez Gene ID: (Human) 3213, (Mouse) 15410, (Rat) 303488

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RNA polymerase II core promoter sequence-specific DNA binding transcription factor activity, sequence-specific DNA binding DNA binding sequence-specific DNA binding

negative regulation of transcription from RNA polymerase II promoter hematopoietic progenitor cell differentiation transcription, DNA-templated regulation of transcription, DNA-templated multicellular organism development anterior/posterior pattern specification rhombomere development glossopharyngeal nerve morphogenesis thyroid gland development embryonic skeletal system morphogenesis regulation of neurogenesis cartilage development definitive hemopoiesis face development angiogenesis