HSD17B11 Antibody

What is HSD17B11 and what biological functions does it serve?

HSD17B11, also known as hydroxysteroid (17-beta) dehydrogenase 11 or short-chain dehydrogenase/reductase member 8 (DHRS8), is an enzyme that catalyzes the conversion of 5α-androstan-3α,17β-diol into androsterone, suggesting a critical role in androgen metabolism . It is nearly ubiquitously expressed, with particularly high expression in the lung, eyes, liver, pancreas, intestine, kidney, adrenal gland, heart, testis, ovary, placenta, and sebaceous gland . Recent studies have shown that HSD17B11 is abundantly expressed in human prostate cancer tissue but not in normal prostate, indicating its potential connection with advanced prostate cancer .

What applications can HSD17B11 antibodies be used for?

HSD17B11 antibodies can be applied in multiple experimental techniques:

The antibody has been validated for reactivity with human, mouse, and rat samples, making it versatile for comparative studies across these species .

What tissue samples have shown positive results with HSD17B11 antibody detection?

Positive Western Blot detection has been confirmed in:

• Mouse pancreas tissue

• Rat pancreas tissue

Positive Immunohistochemistry detection has been confirmed in:

• Human kidney tissue

• Human brain tissue

• Human ovary tissue

These validated tissues provide excellent positive controls for researchers establishing new protocols.

What are the optimal conditions for using HSD17B11 antibody in immunohistochemistry?

For optimal immunohistochemistry results with HSD17B11 antibody:

• Use sections of recommended tissue types (kidney, brain, or ovary for human samples).

• Perform antigen retrieval using TE buffer at pH 9.0 (preferred method).

• Alternatively, citrate buffer at pH 6.0 can be used for antigen retrieval if TE buffer yields suboptimal results.

• Use antibody at a dilution range of 1:20-1:200, optimizing for your specific tissue sample.

• Include appropriate negative controls (omission of primary antibody) and positive controls (tissues known to express HSD17B11).

• Counterstain and mount according to standard laboratory protocols .

It is strongly recommended that the antibody be titrated in each testing system to obtain optimal results, as the optimal dilution may be sample-dependent .

How should HSD17B11 antibody be stored to maintain optimal activity?

To preserve antibody activity:

• Store unopened antibody at -20°C. When stored properly, it remains stable for one year after shipment.

• For -20°C storage, aliquoting is unnecessary for the standard 20μl sizes that contain 0.1% BSA.

• The antibody is supplied in PBS with 0.02% sodium azide and 50% glycerol at pH 7.3.

• Avoid repeated freeze-thaw cycles that can denature the antibody and reduce its effectiveness .

These storage conditions ensure that the antibody maintains its specific binding properties and provides consistent results across experiments.

How is HSD17B11 transcriptionally regulated in cancer cells?

The transcriptional regulation of HSD17B11 in prostate cancer cells involves several key elements:

• The region -107/+18 of the HSD17B11 gene has been identified as the minimal promoter sufficient for activity in prostate cancer cells.

• This critical promoter region contains consensus binding sites for multiple transcription factors, including C/EBP, Sp1, GATA, and NF-κB.

• Mutagenesis analyses have demonstrated that the C/EBP and Sp1 binding sites are essential for promoter activity.

• Experiments have shown that ectopic expression of Sp1 and C/EBPα specifically upregulates HSD17B11 expression in prostate cancer cell lines.

• DAPA (DNA affinity precipitation assay) and ChIP (Chromatin Immunoprecipitation) assays have confirmed the specific recruitment of Sp1 and C/EBPα to the HSD17B11 promoter .

These findings provide valuable insights for researchers investigating transcriptional mechanisms and potential therapeutic targets in hormone-dependent cancers.

What is the relationship between HSD17B11 and non-coding RNAs in cancer progression?

Recent studies have identified a complex relationship between HSD17B11-related non-coding RNAs and cancer:

This relationship between HSD17B11-related non-coding RNAs and cancer progression represents an emerging area for therapeutic targeting and biomarker development.

What are the common challenges when using HSD17B11 antibody in Western blot and how can they be addressed?

Common challenges and solutions for Western blotting with HSD17B11 antibody include:

• Background issues:

  • Ensure thorough blocking (5% non-fat milk or BSA in TBST is recommended)

  • Increase washing steps and duration

  • Use fresh blocking reagents and antibody dilutions

• Weak or no signal:

  • Verify protein expression in your sample (HSD17B11 has a calculated molecular weight of 33 kDa)

  • Use positive control samples (mouse or rat pancreas tissue are recommended)

  • Optimize antibody dilution (starting with 1:1000 and adjusting as needed)

  • Extend primary antibody incubation time (overnight at 4°C may improve results)

• Multiple bands:

  • Ensure sample preparations contain protease inhibitors

  • Confirm specificity using different antibody clones

  • Consider post-translational modifications that may affect protein migration

Following the manufacturer's specific Western blot protocol for this antibody (17301-1-AP) is strongly recommended for optimal results.

How can researchers quantitatively measure HSD17B11 levels in biological samples?

For quantitative measurement of HSD17B11 in biological samples, researchers can utilize ELISA techniques:

• Commercial sandwich enzyme immunoassay kits are available for quantitative measurement of HSD17B11 in human serum, plasma, cell culture supernatants, tissue homogenates, and other biological fluids.

• These assays employ antibodies specific for Human HSD17B11 pre-coated onto microplates. The general methodology involves:

  • Adding standards and samples to wells, where any HSD17B11 present binds to the immobilized antibody

  • Washing away unbound substances

  • Adding a detection antibody specific for HSD17B11

  • Adding enzyme conjugate after washing

  • Adding substrate solution to develop color proportional to the amount of bound HSD17B11

  • Measuring absorbance after stopping the color development

• For mRNA quantification, researchers can use RT-qPCR with the following primers:

  • Sense strand: 5'-TGCAATGACGAAGAATAACC-3'

  • Antisense strand: 5'-TTGTAAGGCAGCCAGTTC-3'

  • Reference genes such as GAPDH and ACTB should be used for normalization

These quantitative approaches enable precise measurement of HSD17B11 expression in experimental systems, facilitating comparative studies across different conditions or treatment regimens.

What is the potential role of HSD17B11 in non-cancer pathologies?

While HSD17B11 has been extensively studied in cancer contexts, particularly prostate and colorectal cancers, emerging research suggests potential roles in other pathological conditions:

• Given its expression in steroidogenic cells, including Leydig and granulosa cells, HSD17B11 may have implications for reproductive disorders and fertility issues.

• Its high expression in multiple tissues (lung, eyes, liver, pancreas, intestine, kidney, etc.) suggests potential involvement in tissue-specific metabolic or inflammatory conditions.

• As an enzyme involved in androgen metabolism, HSD17B11 may have relevance to endocrine and metabolic disorders beyond cancer .

Researchers investigating these areas should consider tissue-specific expression patterns and enzyme activity in their experimental designs.

How can gene editing tools be applied to study HSD17B11 function?

Modern gene editing approaches offer powerful tools for investigating HSD17B11 function:

• CRISPR-Cas9 knockout strategies:

  • Target sequences can be designed to disrupt the HSD17B11 gene

  • The gene sequence information (GenBank Accession Number: BC016367) provides the basis for guide RNA design

  • Knockout cell lines can reveal phenotypic consequences of HSD17B11 loss

• Promoter studies:

  • CRISPR activation (CRISPRa) or inhibition (CRISPRi) systems can be used to modulate transcription

  • The identified promoter region (-107/+18) with C/EBP and Sp1 binding sites provides specific targets

  • Mutations can be introduced in specific transcription factor binding sites to validate their functionality

• Interaction studies:

  • Gene editing can be combined with pulldown assays to identify protein-protein interactions

  • RNA-protein interactions, particularly with the related lnc-HSD17B11-1:1, can be investigated using CRISPR-based techniques

These approaches can significantly advance our understanding of both the basic biology and disease-relevant functions of HSD17B11.