TMX1 / Thioredoxin-related transmembrane protein 1 · IHC design guide

Design Immunohistochemistry for TMX1

Plan chromogenic TMX1 IHC in paraffin sections with the IHC-validated antibody at 1:50–1:200 (datasheet: A07462). Assess cytoplasmic and membranous staining in positive cell populations, including lung macrophages (HPA tissue IHC).

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

Immunohistochemistry protocol sheet for TMX1 (IHC for TMX1): expected localisation Cytoplasmic and membranous in tissue IHC (HPA tissue IHC), antibody A07462, validated IHC image, and IHC protocol steps — Printable **TMX1** IHC protocol sheet — expected localisation Cytoplasmic and membranous in tissue IHC (HPA tissue IHC), antibody A07462, controls and protocol steps. Open the full TMX1 IHC guide →

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TMX1 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 and membranous in tissue IHC (HPA tissue IHC)
	Staining pattern	Glandular cells and lung macrophages; cytoplasmic/membranous (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+4 more · see all

Important caveats

Reasons your staining may differ from the expected pattern.

	Fixation	Keep paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific)
	Caveat	Antibody staining and RNA show medium consistency (HPA tissue IHC)
	Regulation	Thrombin stimulates secretion (UniProt)
	Isoform / epitope	No isoforms; map epitope to residues 27–180 or 204–280 (UniProt)

Section 1

Recommended TMX1 IHC & IF Protocols

The catalog antibody protocol is accompanied by one published TMX1 IHC protocol using paraffin sections of human skin and melanoma (PMC6669928).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Paraffin-embedded rat brain tissue; fixative not specified (datasheet A07462)
Fixation	Image fixative and duration unreported (datasheet A07462); 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% normal serum of the secondary host, 30 min, room temperature (standard)
Primary antibody	Rabbit anti-TMX1, 1:50-1:200 (datasheet A07462)
Primary incubation	Overnight at 4 °C (standard)
Detection	HRP-polymer secondary, DAB chromogen 5–10 min (standard)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	TMX1-positive staining in glandular cells of appendix (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic and membranous expression in several tissues. No signal in the no-primary control.

💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval rule); the published protocol specifies target retrieval for 20 min (PMC6669928).

Section 2

What Is the Expected TMX1 Staining Pattern?

TMX1 is predominantly an endoplasmic reticulum membrane protein with one transmembrane segment at residues 181–203 (UniProt Q9H3N1 topology and localization). In paraffin IHC, expect cytoplasmic and membranous staining in selected cells, including appendix glandular cells and lung macrophages, both reported at medium intensity (HPA tissue IHC). The HPA tissue antibody is Approved, with medium consistency between staining and RNA data (HPA tissue IHC).

What am I looking at on my slide?
Cytoplasmic and membranous staining in appendix or duodenal glandular cells.	This fits the reported medium staining in those cells (HPA tissue IHC) and TMX1’s predominant ER membrane localization (UniProt Q9H3N1 localization). Chromogenic IHC need not resolve the ER as a distinct network (general IHC practice).
Strong staining confined to nuclei, with little cytoplasmic signal.	This does not match the reported tissue IHC profile (HPA tissue IHC). Check specificity and section background before scoring it as TMX1. Nucleolar localization is separately supported in ICC-IF, so an IF nucleolar signal alone is not grounds to reject that assay (HPA subcellular ICC-IF).
Prominent staining in kidney glomerular cells or liver cholangiocytes.	Neither cell population had detectable staining in the cited HPA images (HPA tissue IHC). Investigate nonspecific binding or endogenous detection activity (general IHC practice). These cell-specific observations do not establish that an entire kidney or liver sample lacks TMX1 (HPA tissue IHC; UniProt Q9H3N1 tissue specificity).
Widespread, hazy chromogen that obscures cell boundaries.	A haze cannot be assigned confidently to TMX1’s reported cytoplasmic and membranous pattern (HPA tissue IHC). Review negative controls, blocking, washing and chromogen development for assay background (general IHC practice).
No staining in appendix glandular cells or lung macrophages.	Those cell populations showed medium staining in HPA tissue IHC (HPA tissue IHC). A negative result warrants checking tissue preservation, retrieval and detection with appropriate controls (general IHC practice); HPA provides no TMX1-specific fixation-sensitivity finding.

💡Expected TMX1 appearanceCall a positive when selected cells show interpretable cytoplasmic and membranous chromogen, with medium staining plausible in appendix glandular cells or lung macrophages (HPA tissue IHC); isolated nuclear staining or diffuse haze calls for specificity and background checks (HPA tissue IHC; general IHC practice).

How each factor affects the staining
Compartment and topology	TMX1 is predominantly on the ER membrane and spans the membrane at residues 181–203 (UniProt Q9H3N1 topology and localization). Score a cellular cytoplasmic or membranous pattern in IHC rather than requiring resolution of an ER network (HPA tissue IHC; general IHC practice).
Choice of positive and comparator cells	Appendix glandular cells and lung macrophages are medium-staining examples; adipocytes and esophageal squamous cells were not detected (HPA tissue IHC). Use the specified cell populations as comparators, since the HPA calls are cell-specific (HPA tissue IHC).
Strength of validation	The tissue IHC profile is Approved and has medium consistency with RNA expression data (HPA tissue IHC). HPA003085 is Approved for IHC, while the supplied antibody record does not designate it Enhanced (HPA antibodies). Treat an unexpected pattern as a result to verify, not as established TMX1 distribution.
Expression versus sampled cells	UniProt describes TMX1 as ubiquitous and highly expressed in kidney and liver (UniProt Q9H3N1 tissue specificity). HPA reports no detection specifically in glomerular cells and cholangiocytes (HPA tissue IHC). Those statements concern different levels of sampling; neither predicts staining in every cell.
Secretion context	UniProt reports extracellular secretion following thrombin stimulation (UniProt Q9H3N1 localization). That context does not establish extracellular staining in routine paraffin tissue sections; interpret extracellular chromogen against controls and the reported tissue IHC pattern (HPA tissue IHC; general IHC practice).
IF/ICC: what pattern should I expect?	HPA supports mainly ER localization and additional nucleolar localization in ICC-IF (HPA subcellular ICC-IF). This is a separate imaging observation; the IHC decision here uses the cytoplasmic and membranous tissue profile (HPA tissue IHC).

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
Expected positive cells show no chromogen.	The run may have failed, or the selected cells may be poorly preserved (general IHC practice). HPA reports medium staining in appendix glandular cells and lung macrophages (HPA tissue IHC); it does not establish TMX1-specific fixation sensitivity.	Confirm the intended cells are present, check a same-run positive control, and review retrieval and detection against the chosen IHC procedure (general IHC practice). Do not infer TMX1 loss from a failed control.
Only nuclei stain strongly in tissue IHC.	Nuclear-only tissue staining diverges from HPA’s cytoplasmic and membranous profile (HPA tissue IHC). Nucleoli are an additional location in ICC-IF, a different assay (HPA subcellular ICC-IF).	Compare with a negative reagent control and an HPA-positive tissue cell population; review primary-antibody specificity and counterstain interpretation (HPA tissue IHC; general IHC practice).
Unexpected cell populations stain prominently.	Cross-reactivity or endogenous detection activity may contribute (general IHC practice). Glomerular cells and cholangiocytes were not detected in HPA’s sampled tissue images (HPA tissue IHC).	Identify the stained cell type before comparing it with HPA; examine reagent controls and, for enzyme detection, the relevant endogenous-activity block (general IHC practice). Avoid a whole-organ negative call from one cell type (HPA tissue IHC).
Diffuse haze obscures the expected cells.	Nonspecific reagent binding or excess chromogen development can obscure compartment assignment (general IHC practice). The HPA tissue profile is cytoplasmic and membranous (HPA tissue IHC).	Inspect negative controls, then review blocking, washes and development time within the established IHC workflow (general IHC practice). Score TMX1 only where cellular staining can be distinguished from background (HPA tissue IHC).
A kidney or liver section seems inconsistent with published expression.	UniProt reports high expression in those organs, while HPA reports no detection in kidney glomerular cells and liver cholangiocytes specifically (UniProt Q9H3N1 tissue specificity; HPA tissue IHC).	Annotate the actual compartment and cell population before comparing results. Use a documented HPA-positive cell population and same-run controls to assess assay performance (HPA tissue IHC; general IHC practice).
Extracellular staining appears alongside weak cellular signal.	Thrombin-associated secretion is reported by UniProt, but the supplied tissue IHC profile describes cytoplasmic and membranous expression (UniProt Q9H3N1 localization; HPA tissue IHC). The extracellular signal has no established expectation for these sections.	Check whether extracellular color persists in negative controls, then report cellular and extracellular observations separately (general IHC practice). Do not assign extracellular color to TMX1 solely from the secretion annotation (UniProt Q9H3N1 localization).

Source: UniProt Q9H3N1 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing TMX1 staining

Immunohistochemistry of paraffin-embedded rat brain using TMX1 antibody at dilution of 1:100 .Perform microwave antigen retrieval with 10 mM PBS buffer pH 7.2 before commencing with IHC staining protocol.

A07462 · IHCImmunohistochemistry of paraffin-embedded rat brain using TMX1 antibody at dilution of 1:100 .Perform microwave antigen retrieval with 10 mM

Immunohistochemistry of paraffin-embedded rat spleen using TMX1 antibody at dilution of 1:100 .Perform microwave antigen retrieval with 10 mM PBS buffer pH 7.2 before commencing with IHC staining protocol.

A07462 · IHCImmunohistochemistry of paraffin-embedded rat spleen using TMX1 antibody at dilution of 1:100 .Perform microwave antigen retrieval with 10 m

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000139921-TMX1/tissue/cerebral+cortex#imid_1050644

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000139921-TMX1/tissue/colon#imid_1050578

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000139921-TMX1/tissue/liver#imid_1050563

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000139921-TMX1/tissue/kidney#imid_1050564

HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA003085; green = TMX1, blue = nucleus, red = microtubules.

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

Sample controls for TMX1 IHC & IF

🧪Run appendix first and assess glandular cells for staining (HPA: Medium in appendix glandular cells); use adipose tissue adipocytes as the negative comparison (HPA: Not detected in adipocytes). On the appendix slide, morphologically intact cells without chromogen can provide an internal background reference, but the supplied HPA row does not identify a specific negative cell type (HPA: appendix glandular cells, Medium).

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

Negative control tissue: Adipose tissue (HPA Not detected)

ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show TMX1 in A-431, U-251MG, U2OS, with annotated localisation: Endoplasmic reticulum (supported) (HPA subcellular).

Technical controls: Include a no-primary, secondary-only control and an isotype control matched to the primary antibody’s host species, clonality, and concentration; use target knockout material or a validated immunizing-peptide block as a biological specificity control (standard IHC practice). For chromogenic appendix IHC, block endogenous peroxidase and check the no-primary slide for residual background; block endogenous biotin if using avidin–biotin detection (standard IHC practice).

⚠️Feasibility: A target-specific fixation window or fixation effect is unreported in the supplied evidence, and the selected A07462 paraffin-section caption does not state its fixative (A07462 tissue-IHC caption). That caption reports microwave retrieval in 10 mM PBS, pH 7.2, before staining at 1:100; it offers a retrieval starting point but does not establish retrieval dependence (A07462 tissue-IHC caption). HPA supports ER localization by ICC-IF, but the supplied evidence does not establish that IF or frozen sections are easier than paraffin IHC (HPA: endoplasmic reticulum, supported); in appendix chromogenic sections, assess endogenous peroxidase background before scoring glandular staining (standard IHC practice).

HPA tissue IHC evidence for TMX1

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium 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	Medium	Protein (IHC)	HPA →
Bone marrow	Hematopoietic cells	Medium	Protein (IHC)	HPA →
Breast	Glandular cells	Medium	Protein (IHC)	HPA →
Caudate	Glial cells	Medium	Protein (IHC)	HPA →
Duodenum	Glandular cells	Medium	Protein (IHC)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
Adipose tissue	Adipocytes	Not detected	Protein (IHC)	HPA →
Esophagus	Squamous epithelial cells	Not detected	Protein (IHC)	HPA →
Gallbladder	Glandular cells	Not detected	Protein (IHC)	HPA →
Heart muscle	Cardiomyocytes	Not detected	Protein (IHC)	HPA →
Kidney	Cells in glomeruli	Not detected	Protein (IHC)	HPA →

Section 3

Advanced TMX1 IHC Tips

Troubleshoot TMX1 chromogenic IHC in paraffin sections using the tissue staining evidence, protein topology, and controls appropriate to each specimen.

Which retrieval conditions should I try when TMX1 staining is weak in paraffin sections?

Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 min (page retrieval rule: cytoplasmic or membrane antigen). If staining remains weak, compare a section retrieved by microwave in 10 mM PBS at pH 7.2, the condition reported for the catalog antibody at 1:100 in paraffin-embedded rat brain (caption: A07462). Keep section thickness, antibody dilution, and chromogen development constant across the comparison (standard IHC practice). Assess whether stronger staining retains a plausible cytoplasmic or membranous pattern, since greater signal alone does not establish specificity (HPA: tissue IHC profile; standard IHC practice).

Can the available evidence identify a fixation condition that preserves TMX1 staining?

TMX1-specific sensitivity to fixation is unknown because the selected rat brain caption reports paraffin embedding but does not state a fixative (caption: A07462). For a controlled starting comparison, document fixation time and process matched samples consistently; 10% neutral buffered formalin is a routine paraffin IHC fixative, not a TMX1-validated condition (standard IHC practice). Apply the same citrate pH 6.0 retrieval and antibody dilution to each sample when assessing fixation effects (page retrieval rule; standard IHC practice). Judge preservation by cell morphology and the expected cytoplasmic or membranous pattern, with a negative control alongside each condition (HPA: tissue IHC profile; standard IHC practice).

What staining pattern is plausible for TMX1, and when should localisation raise concern?

Expect predominantly intracellular staining compatible with endoplasmic reticulum localisation; the tissue survey describes cytoplasmic and membranous expression in several tissues (UniProt Q9H3N1 subcellular location; HPA: tissue IHC profile). TMX1 spans the membrane at residues 181–203, so a membranous component can be plausible, although chromogenic IHC cannot resolve that membrane precisely (UniProt Q9H3N1 topology; standard IHC practice). Mitochondria-associated ER localisation is also reported, and secretion follows thrombin stimulation; neither observation predicts diffuse extracellular staining in an unstimulated section (UniProt Q9H3N1 subcellular location). Review strong nuclear-only or uniform extracellular signal against tissue morphology and controls before assigning it to TMX1 (standard IHC practice).

How should I troubleshoot an unexpected pattern when the antibody epitope is unspecified?

First confirm the antibody's mapped epitope, if available, rather than assuming that every TMX1 region is equally accessible after processing (standard IHC practice). The record lists 0 isoforms, a signal peptide at residues 1–26, a thioredoxin domain at 27–132, and a transmembrane segment at 181–203 (UniProt Q9H3N1 sequence features). An epitope within the cleaved signal peptide would require particular scrutiny, while an epitope on either side of the membrane may respond differently to retrieval; the catalog antibody's epitope is not specified here (UniProt Q9H3N1 processing and topology; standard IHC practice). Compare plausible localisation and control sections before attributing a changed chromogenic pattern to an isoform (HPA: tissue IHC profile; UniProt Q9H3N1 isoforms).

How can I use IF to check a disputed TMX1 IHC pattern?

Use IF as a separate localisation check: HPA reports supported ER localisation, with additional nucleolar localisation, in its cell imaging data (HPA: subcellular). Multiplex TMX1 with a validated marker for the expected cell population, such as the macrophage population in lung, and inspect colocalisation at the single-cell level (HPA: Medium staining in lung macrophages; standard IF practice). Select a fluorophore in a channel with low measured tissue autofluorescence, and include single-stain and no-primary controls to assess bleed-through and background (standard IF practice). Choose permeabilisation according to whether the mapped epitope lies before or after the 181–203 transmembrane segment, testing access empirically when its orientation is uncertain (UniProt Q9H3N1 topology; standard IF practice).

What should I check when TMX1 chromogenic staining looks diffuse or widespread?

Run a no-primary control and inspect endogenous pigment, tissue folds, and edge staining before changing antibody concentration (standard IHC practice). In a peroxidase-based assay, block endogenous peroxidase and compare chromogen development times on matched sections; these are general DAB workflow checks, not TMX1-specific findings (standard IHC practice). Titrate the catalog antibody around the reported 1:100 rat brain use while holding retrieval and detection constant (caption: A07462; standard IHC practice). Because HPA reports only medium consistency between antibody staining and RNA expression, interpret a broad signal alongside cell identity and the expected cytoplasmic or membranous distribution (HPA: Approved reliability description and tissue IHC profile).

How should I score TMX1 staining across samples with different cell mixtures? ⚠ ANSWER MARKED FOR VERIFICATION

Define the cell population and compartment before scoring, since HPA reports cytoplasmic and membranous staining across several tissues (HPA: tissue IHC profile). For each matched region, record the percentage of positive cells and an H-score using intensity grades 0–3, or report positive-cell density per mm² when cell abundance is the main outcome (standard IHC quantification practice). Normalize the positive count to the number of eligible cells in that region, or use area for density, while keeping section selection, imaging, and thresholds fixed (standard IHC practice). Report cell types separately where possible: lung macrophages stain at a medium level, whereas kidney glomerular cells were not detected in the HPA survey (HPA: lung macrophages; HPA: kidney glomerular cells).

How do I distinguish convincing TMX1 signal from a staining artefact?

A convincing result follows intact cells and a plausible cytoplasmic or membranous distribution, with a weak no-primary control and reproducibility in matched sections (HPA: tissue IHC profile; standard IHC practice). Check the stained cell type against the reference: HPA reports medium staining in lung macrophages and no detection in adipocytes, so a uniformly positive field needs closer review (HPA: lung macrophages; HPA: adipocytes). Exclude section edges, folds, and necrotic areas, then assess whether residual DAB reflects endogenous peroxidase or excessive chromogen development (standard IHC practice). Predominantly nuclear-only staining warrants scrutiny, although IF data report additional nucleolar localisation; the two assays should be interpreted with their respective controls (HPA: subcellular; standard IHC practice).

Boster reagents

Best TMX1 / Thioredoxin-related transmembrane protein 1 IHC Antibodies

The catalog antibody has real IHC images from paraffin-embedded rat brain and spleen (catalog IHC captions); Human, Mouse and Rat reactivity is listed (catalog reactivity). No IF data are supplied (catalog IF images).

Real IHC data Immunohistochemistry of paraffin-embedded rat brain using TMX1 antibody at dilution of 1:100 .Perform microwave antigen retrieval with 10 mM PBS buffer pH 7.2 before commencing with IHC staining protocol.

Anti-TMX1 Antibody

Cat # A07462

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A07462 is the sole card and is listed for IHC (catalog applications). Its IHC images show paraffin-embedded rat brain and spleen at 1:100 with microwave retrieval in 10 mM PBS, pH 7.2 (A07462 IHC captions).

Which to pick: Choose A07462 for tissue IHC in paraffin sections: its rat brain and spleen captions document that processing, while the fixative is unreported (A07462 IHC captions). It is listed as a rabbit antibody reactive with Human, Mouse and Rat, though the supplied IHC images show Rat only; clonality is unreported (catalog host, reactivity and clone; A07462 IHC captions). No SKU is listed for IF/ICC because A07462 has no IF application or IF image in the payload (catalog applications and IF images).

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

References

UniProt Consortium. UniProt entry Q9H3N1 (TMX1_HUMAN, Thioredoxin-related transmembrane protein 1).
Human Protein Atlas. TMX1 tissue IHC expression (reliability: Approved).
Human Protein Atlas. TMX1 subcellular location (ICC-IF): Mainly localized to the endoplasmic reticulum. In addition localized to the nucleoli..
Human Protein Atlas. TMX1 antibody validation summary (1 antibodies).
Thioredoxin related transmembrane protein 1acts as a prognostic indictor and promotes proliferation and TMZ resistance of lower-grade glioma. Scientific reports 2025 — PMC11822014.
Redox signals at the ER-mitochondria interface control melanoma progression. The EMBO journal 2019 — PMC6669928.
Exploiting potential molecular compounds for treating testicular seminoma by targeting immune related genes. Cell communication and signaling : CCS 2024 — PMC11580341.
PubMed PMID:11152479 — UniProt-cited evidence.
PubMed PMID:12975309 — UniProt-cited evidence.
PubMed PMID:14702039 — UniProt-cited evidence.