Recombinant Human Androgen receptor (AR), partial

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Code CSB-EP001975HU
MSDS
Size $224
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  • (Tris-Glycine gel) Discontinuous SDS-PAGE (reduced) with 5% enrichment gel and 15% separation gel.

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Product Details

Purity
Greater than 90% as determined by SDS-PAGE.
Target Names
AR
Uniprot No.
Research Area
Signal Transduction
Alternative Names
AIS; ANDR_HUMAN; Androgen nuclear receptor variant 2; Androgen receptor (dihydrotestosterone receptor; testicular feminization; spinal and bulbar muscular atrophy; Kennedy disease); Androgen receptor; androgen receptor splice variant 4b; AR; AR8; DHTR; Dihydro testosterone receptor; Dihydrotestosterone receptor (DHTR); Dihydrotestosterone receptor; HUMARA; HYSP1; KD; Kennedy disease (KD); NR3C4; Nuclear receptor subfamily 3 group C member 4 (NR3C4); Nuclear receptor subfamily 3 group C member 4; SBMA; SMAX1; Spinal and bulbar muscular atrophy (SBMA); Spinal and bulbar muscular atrophy; Testicular Feminization (TFM); TFM
Species
Homo sapiens (Human)
Source
E.coli
Expression Region
551-919aa
Target Protein Sequence
DYYFPPQKTCLICGDEASGCHYGALTCGSCKVFFKRAAEGKQKYLCASRNDCTIDKFRRKNCPSCRLRKCYEAGMTLGARKLKKLGNLKLQEEGEASSTTSPTEETTQKLTVSHIEGYECQPIFLNVLEAIEPGVVCAGHDNNQPDSFAALLSSLNELGERQLVHVVKWAKALPGFRNLHVDDQMAVIQYSWMGLMVFAMGWRSFTNVNSRMLYFAPDLVFNEYRMHKSRMYSQCVRMRHLSQEFGWLQITPQEFLCMKALLLFSIIPVDGLKNQKFFDELRMNYIKELDRIIACKRKNPTSCSRRFYQLTKLLDSVQPIARELHQFTFDLLIKSHMVSVDFPEMMAEIISVQVPKILSGKVKPIYFHT
Note: The complete sequence including tag sequence, target protein sequence and linker sequence could be provided upon request.
Mol. Weight
62.4kDa
Protein Length
Partial
Tag Info
N-terminal 10xHis-SUMO-tagged and C-terminal Myc-tagged
Form
Liquid or Lyophilized powder
Note: We will preferentially ship the format that we have in stock, however, if you have any special requirement for the format, please remark your requirement when placing the order, we will prepare according to your demand.
Buffer
If the delivery form is liquid, the default storage buffer is Tris/PBS-based buffer, 5%-50% glycerol.
Note: If you have any special requirement for the glycerol content, please remark when you place the order.
If the delivery form is lyophilized powder, the buffer before lyophilization is Tris/PBS-based buffer, 6% Trehalose, pH 8.0.
Reconstitution
We recommend that this vial be briefly centrifuged prior to opening to bring the contents to the bottom. Please reconstitute protein in deionized sterile water to a concentration of 0.1-1.0 mg/mL.We recommend to add 5-50% of glycerol (final concentration) and aliquot for long-term storage at -20°C/-80°C. Our default final concentration of glycerol is 50%. Customers could use it as reference.
Troubleshooting and FAQs
Storage Condition
Store at -20°C/-80°C upon receipt, aliquoting is necessary for mutiple use. Avoid repeated freeze-thaw cycles.
Shelf Life
The shelf life is related to many factors, storage state, buffer ingredients, storage temperature and the stability of the protein itself.
Generally, the shelf life of liquid form is 6 months at -20°C/-80°C. The shelf life of lyophilized form is 12 months at -20°C/-80°C.
Lead Time
3-7 business days
Notes
Repeated freezing and thawing is not recommended. Store working aliquots at 4°C for up to one week.
Datasheet & COA
Please contact us to get it.
Description

A target DNA sequence encoding to the 551-919aa of human Androgen receptor (AR) was fused with an N-terminal 10xHis-SUMO-tag and a C-terminal Myc-tag and then expressed in the E.coli. The resulting protein is a partial-length recombinant AR protein. It got purified by the SDS-PAGE analysis over 90% in purity. The SDS-PAGE of this AR protein gave a 55-66 kDa band. The in-stock protein allows it to reach your lab bench faster. This recombinant AR protein may be applied to generate specific antibodies and in the AR-related signal transduction research.

AR belonging to the steroid hormone nuclear receptor family is a ligand-dependent nuclear transcription factor. Androgen/AR interaction is involved in a broad of biological functions, such as the expression of the male phenotype and the development & maintenance of the reproductive, musculoskeletal, cardiovascular, immune, neural, and hemopoietic systems. De-regulation of AR is associated with some types of cancers in the prostate, lung, and liver, etc.

Customer Reviews and Q&A

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 Q&A
Q:

It says online that the tag is N-terminal 10xHis-SUMO-tagged and C-terminal Myc-tagged:
1. Why is the SUMO-tagged? SUMO-tag is bad for transcription factors.
2. What SUMO-tag was used? SUMO1, SUMO2, SUMO3 or SUMO4?
3. Can this be made without SUMO? Maybe make with His and Myc Tag?

A:
Very nice to receive your inquiry.
1. The SUMO-tag helps protein folding and helps expression of soluble protein.
2. The sequence of our SUMO-tag is as follows:

MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLRRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG


The result of the uniprot blasting is https://www.uniprot.org/uniprot/Q12306, which should be SUMO3.
3. We could try to express this protein using other tag without SUMO, for example, N-terminal 10xHis-tagged and C-terminal Myc-tagged.
Q:

1. Can we just “chop off” the SUMO with a SUMO-specific protease? Has your group done that?
2. If so, which SUMO protease and is it commercially-available?
3. Ulp1 is the protease to cleave after GG in the yeast SUMO3?

A:
Thanks for your response.
1. The other SUMO-tagged proteins we expressed have been successfully digested with our self-produced SUMO enzyme before.
However, we haven't tried to removed tag from this fusion protein using SUMO enzyme yet. If you have interest, you could have a try.
2. Our self-produced SUMO enzyme is as follows. Code: CSB-EP311619SVG Name: Recombinant Saccharomyces cerevisiae Ubiquitin-like-specific protease 1(ULP1) ,partial
We generally use this protein to remove SUMO-tag. LifeSensors has three SUMO enzymes (SUMO protease1, SUMO Protease 2, and SUMOstar Protease from LifeSensors). Our self-produced SUMO enzyme Ulp1, corresponding to SUMO protease1 from the company LifeSensors.
3. Wikipedia shows "Hydrolysis of the alpha-linked peptide bond in the sequence Gly-Gly-!Ala-Thr-Tyr at the C-terminal end of the small ubiquitin-like modifier (SUMO) propeptide, Smt3".
Here is the link: https://en.wikipedia.org/wiki/Ulp1_peptidase Therefore, GG is the cleavage site of the SUMO enzyme. We used the expression vector pet 28a-sumo to introduce thrombin cleavage sites, where R is the cleavage site (LVPRGS). The customer could also try thrombin cleavage and resection, but we cannot be 100% sure that it can be removed.
The sequence of the fusion protein with pET28a-sumo-10xhis/myc is as below.

MAHHHHHHMSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLRRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGGGSHHHHHHHHHHLVPRGSRT+ Target Protein + AAAEQKLISEEDL-

Q:

What is the stability of this protein? Have there been complaints? I've heard that AR-NTD has instability issues.

A:

This protein has been sold many times, with a total of more than 17.5mg, and only one complaint occurred. The reason is that the protein was delayed in transportation, resulting in the degradation of the protein received by the customer. We supplied the customer with the protein again and the customer said that the experiment can be carried out smoothly. Before long, he ordered the protein again.
At present, the stability of the protein is relatively good, and we will conduct quality control before shipment. In addition, we suggest that customers can order small sizes for testing first.

Target Background

Function
Steroid hormone receptors are ligand-activated transcription factors that regulate eukaryotic gene expression and affect cellular proliferation and differentiation in target tissues. Transcription factor activity is modulated by bound coactivator and corepressor proteins like ZBTB7A that recruits NCOR1 and NCOR2 to the androgen response elements/ARE on target genes, negatively regulating androgen receptor signaling and androgen-induced cell proliferation. Transcription activation is also down-regulated by NR0B2. Activated, but not phosphorylated, by HIPK3 and ZIPK/DAPK3.; Lacks the C-terminal ligand-binding domain and may therefore constitutively activate the transcription of a specific set of genes independently of steroid hormones.; Lacks the C-terminal ligand-binding domain and may therefore constitutively activate the transcription of a specific set of genes independently of steroid hormones.
Gene References into Functions
  1. AR expression heterogeneity is linked to distinct castration/enzalutamide responses in castration-resistant prostate cancer. PMID: 30190514
  2. Androgen receptor positive triple negative breast cancer: Clinicopathologic, prognostic, and predictive features PMID: 29883487
  3. In prostate cancer cells, AR-V7 expression is correlated with drug resistance, as AR-V7 upregulation leads to enhanced proliferation potency of cancer cells, indicating unfavorable prognosis of patients. PMID: 30284554
  4. These findings imply that the deep intronic mutation creating an alternative splice acceptor site resulted in the production of a relatively small amount of wildtype androgen receptor mRNA, leading to partial androgen insensitivity syndrome. PMID: 29396419
  5. AR Germline Mutations and Polymorphisms were associated with Prostate Cancer. PMID: 30139231
  6. GTEE also downregulated the expression of AR and prostate-specific antigen (PSA) in both androgen-responsive and castration-resistant PCa cells. By blocking the SREBP-1/AR axis, GTEE suppressed cell growth and progressive behaviors, as well as activating the caspase-dependent apoptotic pathway in PCa cells PMID: 30301150
  7. Suppressed the expression of androgen receptor. PMID: 29981500
  8. An AR motif of the transactivation domain has been identified that contributes to transcriptional activity by recruiting the C-terminal domain of subunit 1 of the general transcription regulator TFIIF. PMID: 29225078
  9. In LNCaP prostate cancer cells, TSG101 overexpression recruits the androgen receptor (AR) to TSG101-containing cytoplasmic vesicles resulting in reduced AR protein level and AR transactivation activity downregulation. Immunofluorescence microscopy demonstrated that TSG101-decorated cytoplasmic vesicles are associated with late endosomes/lysosomes. PMID: 29859188
  10. Study indicates that both mRNA and protein level of AR increase during prostate cancer (PCa) progression. These levels are even higher in metastatic PCa. Further data suggest that elevation of AR may promote PCa metastasis by induction of EMT and reduction of KAT5. PMID: 30142696
  11. This study aimed to determine the presence and localization of oestrogen receptors (ERs), progesterone receptors (PRs), and androgen receptors (ARs) in both healthy and varicose vein wall cells and their relationship with gender. PMID: 30250632
  12. These findings suggest that CDK11 is involved in the regulation of AR pathway and AR can be a potential novel prognostic marker and therapeutic target for osteosarcoma treatment. PMID: 28262798
  13. We use CPRC prostate cancer model and demonstrate that endothelial cells secrete large amount of CCL5 and induces autophagy by suppressing AR expression in prostate cancer cell lines. Consequently, elevated autophagy accelerates focal adhesions proteins disassembly and promoted prostate cancer invasion. Inhibition of both CCL5/CCR5 signaling and autophagy significantly reduces metastasis in vivo PMID: 30200999
  14. Overexpression of nuclear AR-V7 protein identifies a subset of tumors with remarkably aggressive growth characteristics among clinically and histologically high-risk patients at the time of radical prostatectomy. PMID: 29198908
  15. Study defines AR ligand-binding domain homodimerization as an essential step in the proper functioning of this important transcription factor. Dimerization surface harbours over 40 previously unexplained androgen insensitivity syndromes and prostate cancer-associated point mutations. PMID: 28165461
  16. Loss of AR expression was found in the nucleus of penile cancer cells when compared to normal tissues. Cytoplasmic AR immunostaining was observed in a significant number of these cases and was related with poor prognosis and shorter overall survival. PMID: 30099587
  17. The AR polymorphism is associated with POR risk, patients with repeats greater than 22 show a higher risk. Our data suggest that AR genotype could play a role in natural ovarian aging. PMID: 29886316
  18. In all, these data suggest that Aurora A plays a pivotal role in regulation of Androgen receptor variant 7 expression and represents a new therapeutic target in castrate-resistant prostate cancer. PMID: 28205582
  19. The meta-analysis showed that short CAG and GGN repeats in androgen receptor gene were associated with increased risk of prostate cancer, especially in Caucasians. PMID: 28091563
  20. Knockdown of beta-Klotho produced the opposite effects. In conclusion, beta-Klotho inhibits EMT and plays a tumorsuppressive role in prostate cancer (PCa) , linking FGF/FGFR/beta-Klotho signaling to the regulation of PCa progression. PMID: 29749458
  21. The interaction of AR and SP1 contributes to regulate EPHA3 expression. PMID: 29917167
  22. DHX15 regulates androgen receptor (AR) activity by modulating E3 ligase Siah2-mediated AR ubiquitination independent of its ATPase activity promoting prostate cancer progression. PMID: 28991234
  23. The interaction of Nanog with the AR signaling axis might induce or contribute to Ovarian cancer stem cells regulation. In addition, androgen might promote stemness characteristics in ovarian cancer cells by activating the Nanog promoter PMID: 29716628
  24. a significant subset of endometrial cancers express androgen receptor especially a serous cancers. PMID: 29747687
  25. Letter: eradication of androgen receptor amplification, PSA decline, and clinical improvement with high dose testosterone therapy. PMID: 28040353
  26. The results in this meta-analysis indicated that AR CAG and GGN repeat polymorphisms may be an important pathogenesis of cryptorchidism. PMID: 29044734
  27. the inverse relation observed between bone cell activity and tumor cell AR activity in prostate cancer bone metastasis may be of importance for patient response to AR. PMID: 29670000
  28. Length variations of (CAG)n and (GGC)n polymorphism in the transactivation domain of AR, significantly influence hormonal profile, semen parameters, and sexual functions of asthenospermic subjects by down regulating the expression of AR mediating signaling. PMID: 29083935
  29. Data suggest that somatic mosaicism in AR can cause partial androgen insensitivity syndrome. [CASE REPORT] PMID: 29267169
  30. These results identify HoxB13 as a pivotal upstream regulator of AR-V7-driven transcriptomes that are often cell context-dependent in CRPC, suggesting that HoxB13 may serve as a therapeutic target for AR-V7-driven prostate tumors. PMID: 29844167
  31. TRX1 is an actionable castration-resistant prostate cancer therapeutic target through its protection against AR-induced redox stress. PMID: 29089489
  32. these findings reveal AR-genomic structural rearrangements as important drivers of persistent AR signalling in castration-resistant prostate cancer. PMID: 27897170
  33. AR+ was associated with lower breast cancer mortality in the overall study population ( estrogen receptor-negative). PMID: 28643022
  34. nuclear COBLL1 interacts with AR to enhance complex formation with CDK1 and facilitates AR phosphorylation for genomic binding in castration-resistant prostate cancer model cells. PMID: 29686105
  35. A variety of AR mutants are induced under selective pressures of AR pathway inhibition in castration resistant prostate cancer which remain sensitive to the inhibitor darolutamide. PMID: 28851578
  36. c.3864T>C AR novel mutation is responsible for complete androgen insensitivity syndrome [case report] PMID: 29206494
  37. The Spinal and bulbar muscular atrophy is caused by the expansion of a CAG/glutamine tract in the amino-terminus of the androgen receptor PMID: 29478604
  38. Polysomic AR genes show low methylation levels and high AR protein expression on immunohistochemistry PMID: 29802469
  39. Oral administration of RAD140 substantially inhibited the growth of AR/ER(+) breast cancer patient-derived xenografts (PDX). Activation of AR and suppression of ER pathway, including the ESR1 gene, were seen with RAD140 treatment. PMID: 28974548
  40. The aims of this study was to evaluate if extreme CAG and GGN repeat polymorphisms of the androgen receptors influence body fat mass, its regional distribution, resting metabolic rate, maximal fat oxidation capacity and serum leptin, free testosterone and osteocalcin in healthy adult men PMID: 29130706
  41. The CRISPR/Cas9 system was able to edit the expression of AR and restrain the growth of androgen-dependent prostate cancer cells in vitro, suggesting the potential of the CRISPR/Cas9 system in future cancer therapy. PMID: 29257308
  42. A new mechanism for complete androgen insensitivity syndrome (CAIS). A deep intronic pseudoexon-activating mutation in the intron between exons 6 and 7 of AR, detected in two siblings with CAIS, leads to aberrant splicing of the AR mRNA and insufficient AR protein production. PMID: 27609317
  43. In the current work, we have confirmed that the lead androgen receptor DBD inhibitor indeed directly interacts with the androgen receptor DBD and tested that substance across multiple clinically relevant castration-resistant prostate cancer cell lines PMID: 28775145
  44. Androgen receptor CAG repeat polymorphism is not associated with insulin resistance and type 2 diabetes in Sri Lankan males. PMID: 29202793
  45. AR gene CAG repeat polymorphisms are associated with the increased risk of mild endometriosis PMID: 28915409
  46. ARE full sites generate a reliable transcriptional outcome in AR positive cells, despite their low genome-wide abundance. In contrast, the transcriptional influence of ARE half sites can be modulated by cooperating factors. PMID: 27623747
  47. Targeting the Malat1/AR-v7 axis via Malat1-siRNA or ASC-J9 can be developed as a new therapy to better suppress enzalutamide-resistant prostate cancer progression. PMID: 28528814
  48. High circulating AR-V7 levels predicted resistance to abiraterone and enzalutamide in castration-resistant prostate cancer. PMID: 28818355
  49. Results identified the N-terminal region of AR-V7 (splice variants) that interacts with the diffuse B-cell lymphoma homology (DH) domain of Vav3 which increases its expression in castration-resistant prostate cancer (CRPC). PMID: 28811363
  50. The single nucleotide polymorphism G1733A of the androgen receptor gene is significantly associated with recurrent spontaneous abortions in Mexican patients. PMID: 28707146

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Involvement in disease
Androgen insensitivity syndrome (AIS); Spinal and bulbar muscular atrophy X-linked 1 (SMAX1); Androgen insensitivity, partial (PAIS)
Subcellular Location
Nucleus. Cytoplasm.
Protein Families
Nuclear hormone receptor family, NR3 subfamily
Tissue Specificity
[Isoform 2]: Mainly expressed in heart and skeletal muscle.; [Isoform 3]: Expressed in basal and stromal cells of the prostate (at protein level).
Database Links

HGNC: 644

OMIM: 300068

KEGG: hsa:367

STRING: 9606.ENSP00000363822

UniGene: Hs.76704

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