dhrs7ca Antibody
Product Specs
Constituents: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Target Background
Q&A
What experimental validation methods are critical for confirming DHRS7CA antibody specificity in academic research?
To ensure DHRS7CA antibody specificity in research, investigators should employ a combination of orthogonal validation approaches .
Advanced Note: For zebrafish studies, cross-validate with negative controls (e.g., pre-immune serum) to rule out non-specific binding .
How do I resolve conflicting data between WB and IHC results when using DHRS7CA antibodies?
Discrepancies between WB and IHC often stem from antigen conformational differences or experimental artifacts .
Methodological Insight: Perform sequential validation: Start with WB to confirm antibody functionality, then transition to IHC with optimized protocols .
How can I assess DHRS7CA antibody cross-reactivity in non-target species (e.g., human vs. zebrafish)?
Cross-reactivity evaluation requires comparative immunoblotting and sequence alignment .
Critical Consideration: The Nordic Biosite DHRS7CA antibody is zebrafish-specific (UniProt: A8E5C5) , while human DHRS7 antibodies may target divergent isoforms .
What advanced techniques can enhance DHRS7CA antibody utility in functional studies?
For functional insights, integrate antibody-based assays with gene-editing tools .
Case Study: In cervical cancer research, anti-Trop-2 antibodies (e.g., hRS7) demonstrated enhanced ADCC when combined with IL-2, suggesting DHRS7CA antibodies could similarly be tested in immune modulation studies .
How should I interpret unexpected bands in DHRS7CA Western Blots?
Unexpected bands indicate post-translational modifications (PTMs) or cross-reactivity .
| Band Type | Investigation Strategy |
|---|---|
| Higher Molecular Weight | Test for phosphorylation/ubiquitination using phospho-specific antibodies. |
| Lower Molecular Weight | Assess proteolytic cleavage via calpain or caspase inhibitors. |
| Non-specific Smearing | Optimize SDS-PAGE conditions (e.g., reducing agents, acrylamide concentration). |
Best Practice: Perform peptide blocking experiments to confirm epitope specificity . Incubate antibody with excess immunogen peptide to abolish signal.
What are the limitations of DHRS7CA antibodies in developmental biology studies?
Zebrafish-specific DHRS7CA antibodies face challenges in cross-species translation and dynamic expression tracking .
Data Table: DHRS7CA Antibody Characteristics by Source
| Supplier | Reactivity | Applications | Immunogen | Key Reference |
|---|---|---|---|---|
| Nordic Biosite | Zebrafish | WB, ELISA | Recombinant zebrafish DHRS7CA | |
| Elabscience | Human | WB, IHC | Human DHRS7 fusion protein |
Can DHRS7CA antibodies be used for immunoprecipitation (IP) or mass spectrometry (MS)-based workflows?
IP/MS applications require high-affinity antibodies and optimized protocols .
| Protocol Step | Recommendation |
|---|---|
| IP Buffer | Use mild detergents (e.g., NP-40) to preserve protein complexes. |
| Antibody Crosslinking | Avoid crosslinking unless necessary; DHRS7CA may aggregate under harsh conditions. |
| MS Preparation | Use on-bead digestion with trypsin after IP to minimize protein loss. |
Caveat: Polyclonal antibodies may pull down non-specific proteins due to epitope redundancy. Validate hits with reciprocal co-IP or CRISPR KO .
How should I design a DHRS7CA antibody validation strategy for a multi-omics study?
Integrate antibody performance metrics with omics data correlation .
| Validation Tier | Assay | Output |
|---|---|---|
| Tier 1 (Basic) | WB + IHC | Confirm presence/localization in target tissues. |
| Tier 2 (Advanced) | ChIP-Seq + RNA-seq | Link DHRS7CA binding to gene expression changes. |
| Tier 3 (Functional) | CRISPR KO + antibody-based assays | Validate antibody specificity in loss-of-function models. |
Workflow Example:
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WB/IHC: Confirm DHRS7CA expression in zebrafish embryos.
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ChIP-Seq: Map DHRS7CA binding sites genome-wide.
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RNA-seq: Identify downstream targets for functional annotation .
What are the implications of DHRS7CA antibody cross-reactivity with human DHRS7?
Potential cross-reactivity complicates species-specific studies but enables comparative biology .
| Scenario | Action | Risk |
|---|---|---|
| Human DHRS7 Antibody | Test zebrafish lysates to confirm no cross-reactivity. | False positives in zebrafish models. |
| Zebrafish DHRS7CA Antibody | Validate human reactivity via WB to avoid missing conserved epitopes. | Limited utility in human studies. |
Case Insight: The Elabscience human DHRS7 antibody (E-AB-52605) shows no zebrafish reactivity , while Nordic Biosite’s DHRS7CA antibody is zebrafish-specific .
How can I optimize DHRS7CA antibody performance in challenging samples (e.g., paraffin-embedded tissues)?
Optimization requires antigen retrieval (AR) tuning and detection system selection .
| Parameter | Recommendation |
|---|---|
| AR Method | Use citrate buffer (pH 6.0) or EDTA (pH 8.0) for epitope exposure. |
| Primary Incubation | Extend to 4°C overnight to enhance signal in low-abundance samples. |
| Secondary Antibody | Use HRP-conjugated anti-rabbit IgG with tyramide amplification for IHC. |
