PRDX5 / Peroxiredoxin-5, mitochondrial · IHC design guide

Design Immunohistochemistry for PRDX5

Plan chromogenic IHC for PRDX5 in paraffin sections using abundant cytoplasmic staining in glandular cells as a reference (HPA tissue IHC). Consider its mitochondrial and cytoplasmic/peroxisomal isoforms when interpreting localisation (UniProt).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use

Immunohistochemistry protocol sheet for PRDX5 (IHC for PRDX5): expected localisation Cytoplasmic tissue staining (HPA tissue IHC), antibody PB9384, validated IHC image, and IHC protocol steps — Printable **PRDX5** IHC protocol sheet — expected localisation Cytoplasmic tissue staining (HPA tissue IHC), antibody PB9384, controls and protocol steps. Open the full PRDX5 IHC guide →

Navigate to: Protocols Staining pattern Controls Antibodies

PRDX5 Immunohistochemistry Experimental Design Guide

Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.

Must know before staining

	Expected localisation	Cytoplasmic tissue staining (HPA tissue IHC)
	Staining pattern	General cytoplasmic staining, abundant in glandular cells (HPA tissue IHC)
	Antigen retrieval	Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen)
	Positive control ⓘ	Appendix+4 more · see all
	Negative control ⓘ	Adipose tissue

Important caveats

Reasons your staining may differ from the expected pattern.

	Fixation	Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific)
	Caveat	Isoforms occupy different compartments (UniProt)
	Regulation	No specific expression regulator reported (UniProt)
	Isoform / epitope	4 isoforms include mitochondrial and cytoplasmic/peroxisomal forms (UniProt)

Section 1

Recommended PRDX5 IHC & IF Protocols

Compare the catalog antibody’s IHC-P protocol with two published PRDX5 chromogenic IHC protocols (PMC4985551; PMC12848523).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Tissue sections; selected-image fixative not specified (standard IHC workflow)
Fixation	Image fixative and duration unreported (datasheet PB9384); verify before use.
Sectioning	4–5 µm sections on charged slides (standard)
Deparaffinisation	Xylene, graded ethanol series to water (standard)
Antigen retrieval	Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% goat serum (datasheet PB9384)
Primary antibody	Rabbit anti-PRDX5, 0.5-1μg/ml (datasheet PB9384)
Primary incubation	Overnight at 4 °C (datasheet PB9384)
Detection	Streptavidin-biotin complex (SABC), DAB chromogen (datasheet PB9384)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	PRDX5-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression, highly abundant in most glandular cells. No signal in the no-primary control.

💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page antigen retrieval rule); neither cited PRDX5 method specifies retrieval conditions.

Section 2

What Is the Expected PRDX5 Staining Pattern?

PRDX5 should stain mainly in the cytoplasm of many tissues, especially glandular cells; HPA reports high glandular staining and rates its tissue IHC evidence Enhanced (HPA: tissue IHC). Mitochondrial, cytoplasmic and peroxisomal pools are consistent with its reported locations, and PRDX5 has no transmembrane segment (UniProt P30044: subcellular location and topology).

What am I looking at on my slide?
Cytoplasmic staining is strong in appendix or colon glandular cells (HPA: High in both cell populations).	This matches the reported tissue pattern (HPA: general cytoplasmic expression; High in glandular cells). Score the glandular cells separately from surrounding tissue so their signal is not obscured by a section-wide average (standard IHC practice).
Staining is confined to nuclei or outlines plasma membranes, with little cytoplasmic signal.	That distribution conflicts with the reported cytoplasmic IHC pattern and lacks support from PRDX5's listed locations or topology (HPA: tissue IHC; UniProt P30044: location and no transmembrane segment). Check morphology and controls before interpreting it as PRDX5 (standard IHC practice).
Adipocytes stain strongly while established positive cells in the same run stain weakly.	Adipocytes are reported as Not detected, whereas appendix and colon glandular cells are High (HPA: tissue IHC). Consider cross-reactivity or endogenous detection activity; the unexpected pattern alone does not distinguish those causes (standard IHC practice).
Brown signal spreads across tissue structures or appears in the negative control.	A diffuse deposit that does not follow cells is difficult to score as PRDX5 (standard IHC practice). Check the negative control, blocking and detection reagents, and chromogen development before assigning a cellular pattern (standard IHC practice).
No staining appears in appendix or colon glandular cells (HPA: High in both).	An absent signal in these expected positive populations makes a negative test result uninterpretable until the run is checked (HPA: tissue IHC; standard IHC practice). Review antibody dilution, retrieval conditions and detection performance without assuming a PRDX5-specific fixation effect (standard IHC practice).

💡Expected PRDX5 appearanceA convincing positive result shows strong cytoplasmic staining in appendix or colon glandular cells (HPA: High; general cytoplasmic expression), while isolated nuclear outlines or strong adipocyte staining warrant investigation rather than a positive PRDX5 call (UniProt P30044: location; HPA: adipocytes Not detected).

How each factor affects the staining
Cell population and tissue selection	Appendix and colon glandular cells, kidney collecting ducts, and lung type II alveolar cells are reported High (HPA: tissue IHC). Hepatocytes are Low and adipocytes Not detected (HPA: tissue IHC); compare matched cell populations when judging intensity (standard IHC practice).
Intracellular location	UniProt lists mitochondria, cytoplasm and peroxisome matrix (UniProt P30044: subcellular location). HPA describes general cytoplasmic tissue IHC and supported mitochondrial ICC-IF (HPA: tissue IHC; subcellular). Chromogenic IHC alone need not resolve each organelle (standard IHC practice).
Isoforms and processing	Four isoforms include mitochondrial and cytoplasmic plus peroxisomal forms, and the mitochondrial chain spans residues 53–214 (UniProt P30044: isoforms and processing). Without an epitope assignment in these payloads, staining cannot be assigned to one isoform or processing state.
Topology and tissue evidence	PRDX5 has no transmembrane segment (UniProt P30044: topology). HPA rates tissue IHC Enhanced and reports high consistency between antibody staining and RNA expression (HPA: tissue IHC reliability); that supports the reported tissue pattern without validating every unusual stain.
IF/ICC Q: What localisation should a separate IF assessment expect?	A: Mitochondrial localisation is supported in HPA ICC-IF images from MCF-7, U2OS and U-251MG (HPA: subcellular ICC-IF). This is an IF localisation reference; interpret paraffin-section chromogenic IHC using the reported tissue pattern (HPA: tissue IHC).

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
Known positive glandular cells show no signal (HPA: appendix and colon High).	The run may have insufficient detection or unsuitable antibody or retrieval conditions (standard IHC practice); these payloads do not establish PRDX5-specific retrieval requirements.	Confirm the positive control, antibody dilution, retrieval settings and detection reagents, then repeat the run under a documented IHC procedure (standard IHC practice).
Signal is predominantly nuclear or sharply membranous.	The pattern disagrees with HPA cytoplasmic tissue staining and UniProt's listed locations and topology (HPA: tissue IHC; UniProt P30044). Artefact or nonspecific staining is possible (standard IHC practice).	Compare an expected positive cell population and the negative control; reassess cell boundaries and antibody specificity before scoring the unusual compartment (standard IHC practice).
Adipocytes show strong staining.	HPA reports adipocytes as Not detected (HPA: tissue IHC). Cross-reactivity or endogenous detection activity is possible, but the stain alone cannot identify which (standard IHC practice).	Check detection-only controls and inspect whether staining follows adipocyte morphology; compare with High glandular cells processed in the same run (standard IHC practice; HPA: tissue IHC).
Broad brown background obscures cellular detail.	Excess detection signal, inadequate blocking or chromogen overdevelopment can create background in chromogenic IHC (standard IHC practice); this pattern is not a reported PRDX5 distribution (HPA: tissue IHC).	Inspect negative controls, review blocking and detection steps, and reduce chromogen development as indicated by the controls (standard IHC practice).
Hepatocytes stain less intensely than nearby positive control tissue.	Low hepatocyte staining is reported by HPA, while appendix and colon glandular cells are High (HPA: tissue IHC). Lower liver signal alone does not establish assay failure.	Judge liver cells against the reported Low category and verify performance with a High glandular control rather than requiring equal intensity (HPA: tissue IHC; standard IHC practice).
A tissue section shows mixed strong and weak cell populations.	PRDX5 is widely expressed, but HPA reports distinct levels by cell type, including High collecting ducts and Low smooth muscle cells (UniProt P30044: tissue specificity; HPA: tissue IHC).	Identify and score each cell population separately; use the matching HPA cell-level expectation before calling weak cells negative or strong cells nonspecific (HPA: tissue IHC; standard IHC practice).

Source: UniProt P30044 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing PRDX5 staining

PB9384 · IHCAnti-Peroxiredoxin 5 vendor antibody, PB9384, IHC(P) IHC(P): Human Lung Cancer Tissue

PB9384 · IHCAnti-Peroxiredoxin 5 vendor antibody, PB9384, IHC(P) IHC(P): Rat Intestine Tissue

IF analysis of Peroxiredoxin 5 using anti-Peroxiredoxin 5 antibody (PB9384). Peroxiredoxin 5 was detected in immunocytochemical section of HELA cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-Peroxiredoxin 5 Antibody (PB9384) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

PB9384 · IFIF analysis of Peroxiredoxin 5 using anti-Peroxiredoxin 5 antibody (PB9384). Peroxiredoxin 5 was detected in immunocytochemical section of H

PA1838 · IHCAnti-Peroxiredoxin 5 antibody, PA1838, IHC(P) IHC(P): Human Prostatic Cancer Tissue

PA1838 · IHCAnti-Peroxiredoxin 5 antibody, PA1838, IHC(P) IHC(P): Rat Liver Tissue

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000126432-PRDX5/tissue/cerebral+cortex#imid_15304722

HPA · EsophagusHuman Protein Atlas IHC, Esophagus. Source: https://www.proteinatlas.org/ENSG00000126432-PRDX5/tissue/esophagus#imid_15304686

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000126432-PRDX5/tissue/colon#imid_15304640

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000126432-PRDX5/tissue/liver#imid_15304641

HPA · ICC-IF MCF-7Human Protein Atlas immunofluorescence, MCF-7, antibody HPA037915; green = PRDX5, blue = nucleus, red = microtubules.

← swipe / scroll for more images →

Real IHC / IF data behind the staining-pattern expectations in this section.

Sample controls for PRDX5 IHC & IF

🧪Run colon first: glandular cells should stain strongly (HPA: High in colon glandular cells). Use adipose tissue as the negative tissue (HPA: Not detected in adipocytes); on the colon slide, cells without specific signal should show only counterstain or background, but should not be assumed to lack PRDX5.

Positive control tissue: Appendix (Glandular cells, HPA High)

Negative control tissue: Adipose tissue (HPA Not detected)

ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PRDX5 in MCF-7, U2OS, U-251MG, with annotated localisation: Mitochondria (supported) (HPA subcellular).

Technical controls: Include no-primary (secondary-only) and host- and clonality-matched isotype controls, plus PRDX5 knockout material or a validated peptide block as a biological specificity control (standard IHC practice). Check colon sections for endogenous peroxidase signal in chromogenic IHC and autofluorescence in IF before scoring (standard IHC/IF practice).

⚠️Feasibility: A PRDX5-specific fixation window and antigen-retrieval dependency are unreported in the supplied evidence; the selected PB9384 tissue-IHC caption also leaves the fixative unreported (selected-SKU caption). Frozen-section performance is unreported, while ICC-IF images exist for MCF-7, U2OS and U-251MG and show supported mitochondrial localization (HPA: subcellular). In colon IHC, inspect luminal material and background against the controls before scoring glandular staining (standard IHC practice; HPA: High in colon glandular cells).

HPA tissue IHC evidence for PRDX5

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.

Positive expression · recommended positive controls

Tissue	Cell type	Level	Evidence	Source
Appendix	Glandular cells	High	Protein (IHC)	HPA →
Bronchus	Ciliated cells (cell body)	High	Protein (IHC)	HPA →
Cerebellum	Synaptic glomeruli - core	High	Protein (IHC)	HPA →
Colon	Glandular cells	High	Protein (IHC)	HPA →
Fallopian tube	Ciliated cells (cell body)	High	Protein (IHC)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
Adipose tissue	Adipocytes	Not detected	Protein (IHC)	HPA →

Section 3

Advanced PRDX5 IHC Tips

Troubleshoot PRDX5 staining in paraffin sections by checking retrieval, cellular distribution and tissue context before comparing signal intensity.

How should I adjust retrieval when PRDX5 staining is weak or uneven?

Start with citrate buffer at pH 6.0 for heat induced retrieval at 95–98 °C for 20 min (page retrieval rule). If staining remains weak, compare a longer exposure or an alternative retrieval buffer on adjacent sections while holding antibody and detection conditions constant (standard IHC practice). Check whether signal improves in glandular cells, which commonly show abundant PRDX5 staining, without a parallel rise in background (HPA tissue IHC profile). Because PRDX5 has no transmembrane segment, a diffuse cytoplasmic pattern alone does not establish that retrieval worked; evaluate cell type and compartment together (UniProt P30044 topology; HPA subcellular).

Could fixation explain weak PRDX5 staining in my paraffin sections?

Target specific fixation sensitivity is unknown: the selected PB9384 tissue IHC caption does not state a fixative, and the supplied evidence gives no PRDX5 fixation comparison (PB9384 tissue IHC caption). Record the fixative, fixation duration and section processing for each specimen before comparing staining (standard IHC practice). When signal differs between batches, stain sections together using citrate at pH 6.0, 95–98 °C for 20 min, with identical detection conditions (page retrieval rule; standard IHC practice). Treat any fixation effect as a hypothesis until matched tissue processed under defined fixation conditions shows a reproducible difference (standard IHC practice).

Which PRDX5 staining pattern is plausible in tissue sections?

Expect predominantly cytoplasmic staining in tissue IHC, with mitochondrial localisation supported by subcellular imaging (HPA tissue IHC profile; HPA subcellular). PRDX5 is also annotated in cytoplasm and peroxisome matrix, so a chromogenic signal need not resolve exclusively into mitochondrial puncta (UniProt P30044 subcellular). Compare the cell body with nuclei, lumina and section edges, and score only anatomically credible cellular staining (standard IHC practice). In a positive control, glandular cells of appendix or colon show high staining, whereas adipocytes are listed as not detected; these contrasts help check localisation and assay background (HPA tissue IHC).

Could an epitope or isoform difference change the PRDX5 IHC pattern?

PRDX5 has 4 annotated isoforms, including mitochondrial and cytoplasmic plus peroxisomal forms, so compartment differences may reflect isoform biology (UniProt P30044 isoforms; UniProt P30044 subcellular). The mitochondrial mature chain spans residues 53–214, and the thioredoxin domain spans 56–214 (UniProt P30044 processing; UniProt P30044 domains). Check the catalog antibody’s stated immunogen or epitope before interpreting staining as specific to one form; that information is not supplied here (standard IHC practice). Document retrieval and compare controls consistently, because the annotated acetylation, succinylation and phosphorylation sites alone do not establish an effect on this antibody’s binding (UniProt P30044 modified residues; standard IHC practice).

How can IF help assess a questionable PRDX5 IHC localisation?

Use IF as a separate localisation check, pairing PRDX5 with a mitochondrial marker and a marker for the expected cell type in the same specimen (HPA subcellular; standard IF practice). Select fluorophores in channels with low measured tissue autofluorescence, and include single colour controls before judging overlap (standard IF practice). Permeabilise sufficiently for antibody access to an intramitochondrial epitope if that is the antibody’s mapped target; epitope accessibility is unknown here, while PRDX5 lacks a transmembrane segment (UniProt P30044 topology; standard IF practice). HPA reports supported mitochondrial localisation from ICC/IF, but agreement with that pattern does not independently validate the catalog antibody in IF (HPA subcellular; standard IF practice).

What should I check when PRDX5 staining appears throughout the section?

First compare staining with a no primary control and inspect whether colour follows tissue edges, folds or damaged areas rather than intact cells (standard IHC practice). Apply a peroxidase block before chromogenic detection and assess nonspecific binding with the usual protein block and washes (standard IHC practice). Widespread cytoplasmic staining can be biologically plausible because PRDX5 is widely expressed and abundant in many glandular cells, so background cannot be judged by distribution alone (UniProt P30044 tissue specificity; HPA tissue IHC profile). Compare adipocytes, listed as not detected, with high staining glandular cells under the same detection conditions before changing antibody concentration (HPA tissue IHC; standard IHC practice).

How should I score PRDX5 IHC across tissue samples? ⚠ ANSWER MARKED FOR VERIFICATION

Define the cell population and compartment before scoring, then record the percentage of positive cells and intensity using an H score with a 0–300 range (standard IHC practice). If cell density differs substantially, report positive cells per mm² alongside the fraction positive, using the same tissue region rules for every slide (standard IHC practice). Normalise comparisons to the number of eligible intact cells or analysed tissue area, and process matched slides with the same retrieval and detection settings (standard IHC practice). Keep glandular and nonglandular populations separate because HPA reports abundant glandular staining and variable levels among other cell types (HPA tissue IHC).

When is apparent PRDX5 positivity more likely to be artefact?

A credible result follows cellular cytoplasm, with mitochondrial localisation supported by ICC/IF, and varies with the sampled cell population (HPA tissue IHC profile; HPA subcellular). Question isolated nuclear or luminal colour, strong section edge staining and signal concentrated in necrotic regions; assess these against intact neighbouring cells and a no primary control (standard IHC practice). Residual endogenous peroxidase can mimic positive chromogenic staining, so confirm the peroxidase block and inspect the control before interpreting weak DAB signal (standard IHC practice). High staining in glandular cells is plausible, while staining of adipocytes deserves closer scrutiny because HPA lists adipocytes as not detected (HPA tissue IHC).

Boster reagents

Best PRDX5 / Peroxiredoxin-5, mitochondrial IHC Antibodies

PB9384 has IHC images from human lung cancer and rat and mouse intestine, plus IF in HELA cells (PB9384 image captions). PA1838 has IHC images from human prostatic cancer and rat liver (PA1838 image captions).

Real IHC data Anti-Peroxiredoxin 5 vendor antibody, PB9384, IHC(P) IHC(P): Human Lung Cancer Tissue

Anti-Peroxiredoxin 5/PRDX5 Antibody ®

Cat # PB9384

View product →

Real IHC data Anti-Peroxiredoxin 5 antibody, PA1838, IHC(P) IHC(P): Human Prostatic Cancer Tissue

Anti-Peroxiredoxin 5/PRDX5 Antibody ®

Cat # PA1838

View product →

PB9384 is listed for IHC and IF/ICC, with IHC images from human lung cancer tissue and rat and mouse intestine, and an IF image from HELA cells (PB9384 catalog applications and image captions). PA1838 is listed for IHC, with images from human prostatic cancer tissue and rat liver (PA1838 catalog applications and image captions).

Which to pick: For paraffin-section IHC, choose either SKU according to the tissue shown in its own IHC images; fixation is unreported (PB9384 and PA1838 IHC(P) captions). Choose PB9384 for IF/ICC because it lists both applications and has an IF cell image (PB9384 catalog applications and IF caption). For cross-species IHC, both list human, mouse and rat reactivity, while PB9384 additionally shows IHC images from all three species (PB9384 and PA1838 catalog reactivity; PB9384 IHC captions).

Each figure is that product's own IHC / IF validation image from its datasheet.

References

UniProt Consortium. UniProt entry P30044 (PRDX5_HUMAN, Peroxiredoxin-5, mitochondrial).
Human Protein Atlas. PRDX5 tissue IHC expression (reliability: Enhanced).
Human Protein Atlas. PRDX5 subcellular location (ICC-IF): Localized to the mitochondria..
Human Protein Atlas. PRDX5 antibody validation summary (3 antibodies).
Extrapancreatic effects of incretin hormones: evidence for weight-independent changes in morphological aspects and oxidative status in insulin-sensitive organs of the obese nondiabetic Zucker rat (ZFR). Physiological reports 2016 — PMC4985551.
Ethyl β-carboline-3-carboxylate targets PRDX5/c-Jun axis for novel therapeutic strategy against cervical cancer. Discover oncology 2025 — PMC12332166.
Lipopolysaccharide Upregulates Neuroinflammation, Oxidative Stress Responses, and Peroxiredoxins in Depression Models. Brain and behavior 2026 — PMC12848523.
Characterization of the Biochemical Recurrence Prediction Ability and Progression Correlation of Peroxiredoxins Family in Prostate Cancer Based on Integrating Single-Cell RNA-Seq and Bulk RNA-Seq Cohorts. Cancer medicine 2025 — PMC12031674.
PubMed PMID:10095767 — UniProt-cited evidence.
PubMed PMID:10514471 — UniProt-cited evidence.
PubMed PMID:10521424 — UniProt-cited evidence.