SEC61G / Protein transport protein Sec61 subunit gamma · IHC design guide

Design Immunohistochemistry for SEC61G

Plan SEC61G paraffin-section IHC with catalog antibody A12389 and chromogenic detection (datasheet A12389). Expect cytoplasmic staining, but interpret its distribution cautiously because HPA rates the tissue IHC evidence as uncertain (HPA tissue IHC).

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

Immunohistochemistry protocol sheet for SEC61G (IHC for SEC61G): expected localisation Cytoplasmic tissue staining (HPA tissue IHC); ER membrane expected (UniProt), antibody A12389, validated IHC image, and IHC protocol steps — Printable **SEC61G** IHC protocol sheet — expected localisation Cytoplasmic tissue staining (HPA tissue IHC); ER membrane expected (UniProt), antibody A12389, controls and protocol steps. Open the full SEC61G IHC guide →

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SEC61G 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); ER membrane expected (UniProt)
	Staining pattern	Cytoplasmic staining in cells across most tissues (HPA tissue IHC)
	Antigen retrieval	EDTA pH 8.0 HIER, heat-mediated (datasheet A12389)
	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 fixation consistent across paraffin sections (standard IHC practice; not target-specific)
	Caveat	Tissue staining has low concordance with RNA data (HPA tissue IHC)
	Regulation	No expression regulation annotated (UniProt)
	Isoform / epitope	No annotated isoforms; epitope position across the membrane matters (UniProt)

Section 1

Recommended SEC61G IHC & IF Protocols

The catalog antibody’s IHC-P protocol (datasheet: A12389) is accompanied by published SEC61G IHC methods for lung adenocarcinoma and oral squamous cell carcinoma (PMC11693863; PMC10603851; PMC13579860).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Paraffin-embedded human colorectal adenocarcinoma tissue; fixative not specified (datasheet A12389)
Fixation	Image fixative and duration unreported (datasheet A12389); verify before use.
Sectioning	4–5 µm sections on charged slides (standard)
Deparaffinisation	Xylene, graded ethanol series to water (standard)
Antigen retrieval	Heat retrieval: EDTA pH 8.0 (datasheet A12389); 20 min, 95–100 °C (standard)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% goat serum (datasheet A12389)
Primary antibody	Rabbit anti-SEC61G, 2-5 μg/ml (datasheet A12389)
Primary incubation	Overnight at 4 °C (datasheet A12389)
Detection	HRP-conjugated secondary, DAB chromogen (datasheet A12389)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	SEC61G-positive staining in glandular cells of appendix (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control.

💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet: A12389); the oral cancer protocol used EDTA pH 9.0 for 30 min (PMC10603851).

Section 2

What Is the Expected SEC61G Staining Pattern?

SEC61G is a single-pass endoplasmic reticulum (ER) membrane protein with residues 33–61 spanning the membrane (UniProt P60059 topology). In paraffin-section IHC, expect cytoplasmic staining in cells such as colon glandular cells, cerebellar Purkinje cells and lung macrophages, each reported at medium intensity (HPA tissue IHC). Treat this as a provisional pattern: HPA rates the tissue IHC reliability Uncertain because staining and RNA expression show low consistency (HPA tissue IHC).

What am I looking at on my slide?
Defined cytoplasmic staining in colon glandular cells, Purkinje cells or lung macrophages.	This fits the reported medium staining in those cells (HPA tissue IHC) and SEC61G's ER membrane location (UniProt P60059). Chromogenic IHC cannot resolve the ER membrane itself; judge the cell and compartment together. Agreement supports an interpretable result, but the HPA reliability rating remains Uncertain (HPA tissue IHC).
Strong staining confined to nuclei, or outlining plasma membranes without a cytoplasmic pattern.	That compartment is discordant with the reported cytoplasmic tissue pattern (HPA tissue IHC) and ER membrane location (UniProt P60059). Consider nonspecific antibody binding, pigment or a detection artefact before scoring it as SEC61G. Reassess against cellular morphology and a matched detection control (general IHC practice).
Staining appears mainly in an unexpected cell type while an expected cell population is clear.	Compare the exact tissue and cell population: colon glandular cells are medium, whereas duodenal glandular cells were not detected in the supplied HPA observations (HPA tissue IHC). Unexpected staining may reflect cross-reactivity or endogenous detection activity (general IHC practice); it does not by itself prove a new SEC61G distribution.
Weak, diffuse color covers tissue, empty spaces and cells alike.	A field-wide deposit without cell boundaries is difficult to reconcile with cell-associated cytoplasmic staining (HPA tissue IHC). Background from incomplete blocking, detection reagents or chromogen development can obscure a real signal (general IHC practice). Interpret intensity only after the background is low enough to distinguish individual cells.
No stain is visible in a run containing colon glandular cells or cerebellar Purkinje cells.	Both cell populations had medium staining in HPA tissue IHC (HPA tissue IHC), so first check tissue preservation, detection controls, antibody use and antigen retrieval as general IHC practice. HPA's Uncertain reliability means a negative result cannot be attributed to one failed step, nor does it establish absent SEC61G.

💡Expected SEC61G appearanceCall a section positive when identifiable cells show cytoplasmic chromogenic staining, with medium intensity plausible in the listed HPA-positive populations (HPA tissue IHC; UniProt P60059 ER membrane); isolated nuclear color or uniform field-wide deposit calls for artefact review (general IHC practice).

How each factor affects the staining
Location and topology	SEC61G resides in the ER membrane and has one transmembrane segment at 33–61, with residues 1–32 cytoplasmic and 62–68 annotated extracellular (UniProt P60059 topology). These annotations guide compartment interpretation. They do not identify the antibody epitope or establish a SEC61G-specific antigen retrieval or permeabilisation condition.
Tissue and cell context	HPA describes cytoplasmic expression in most tissues and low RNA tissue specificity, yet its sampled IHC results differ by cell population (HPA tissue IHC). For example, colon glandular cells were medium, while adipocytes in adipose tissue were not detected (HPA tissue IHC).
Strength of the IHC evidence	The supplied HPA antibody, HPA053196, has an Uncertain IHC rating; HPA reports low consistency between staining and RNA data and says external verification is pending (HPA antibody validation; HPA tissue IHC). Score compartment and cell identity alongside intensity, and keep conclusions provisional when a result relies on this staining evidence alone.
IF/ICC Q&A: what localisation can be expected?	HPA's ICC-IF summary says Membrane, but lists no main location and no cell lines with ICC-IF images (HPA subcellular). UniProt places SEC61G at the ER membrane (UniProt P60059). Those records support an ER membrane expectation; they do not document a cell-specific IF appearance or supply a validated IF protocol.

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
An expected HPA-positive cell population has no detectable chromogen.	A failed staining or detection step is possible (general IHC practice); HPA's Uncertain IHC rating also limits confidence in the expected intensity (HPA tissue IHC).	Check section integrity and detection controls, then compare routine antigen retrieval conditions and antibody use within the established IHC workflow (general IHC practice). Record the tested conditions without treating any one as SEC61G-validated.
Nuclei dominate the result.	Nuclear dominance conflicts with ER membrane localisation (UniProt P60059) and the reported cytoplasmic tissue profile (HPA tissue IHC); nonspecific binding or detection artefact is possible (general IHC practice).	Review morphology and a matched detection control; score cytoplasmic staining separately from nuclear color (general IHC practice).
Color is widespread, hazy or present over tissue-free areas.	General background from blocking, washing or chromogen development can reduce cell-level contrast (general IHC practice).	Check the detection control, wash quality and development time, then reassess whether discrete cytoplasmic staining remains (general IHC practice; HPA tissue IHC pattern).
Unexpected cells stain strongly, especially near blood or inflammatory material.	Cross-reactivity or endogenous detection activity is possible (general IHC practice); HPA observations are cell-specific and have Uncertain reliability (HPA tissue IHC).	Inspect tissue morphology and use appropriate detection controls, including an endogenous-enzyme control when relevant to the chromogen system (general IHC practice).
A tissue listed as HPA-negative shows cytoplasmic color.	HPA reports no detection in selected cells, such as adipocytes in adipose tissue, rather than proving every cell in that section lacks SEC61G (HPA tissue IHC).	Identify which cell type stains, compare it with the exact HPA cell-level observation, and assess detection controls before calling the result discordant (HPA tissue IHC; general IHC practice).
Two sections show different apparent intensity.	Different cell populations have different reported HPA levels, and a chromogenic readout also depends on consistent processing and development (HPA tissue IHC; general IHC practice).	Compare the same cell population under matched staining and imaging conditions; report intensity with the cell identity and note the Uncertain HPA reliability (general IHC practice; HPA tissue IHC).

Source: UniProt P60059 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing SEC61G staining

IHC analysis of SEC61G using anti-SEC61G antibody (A12389). SEC61G was detected in a paraffin-embedded section of human colorectal adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-SEC61G Antibody (A12389) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

A12389 · IHCIHC analysis of SEC61G using anti-SEC61G antibody (A12389). SEC61G was detected in a paraffin-embedded section of human colorectal adenocarc

IHC analysis of SEC61G using anti-SEC61G antibody (A12389). SEC61G was detected in a paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-SEC61G Antibody (A12389) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

A12389 · IHCIHC analysis of SEC61G using anti-SEC61G antibody (A12389). SEC61G was detected in a paraffin-embedded section of human liver cancer tissue.

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000132432-SEC61G/tissue/cerebral+cortex#imid_14751612

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000132432-SEC61G/tissue/colon#imid_14771752

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000132432-SEC61G/tissue/liver#imid_14771753

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000132432-SEC61G/tissue/kidney#imid_14771673

← swipe / scroll for more images →

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

Sample controls for SEC61G IHC & IF

🧪Run colon first: its glandular cells should stain for SEC61G (HPA: Colon glandular cells, Medium); use adrenal gland glandular cells as a tissue negative (HPA: Adrenal gland glandular cells, Not detected). On the colon slide, nonglandular cells that show counterstain without specific DAB signal can serve as provisional internal background, since the supplied HPA row does not identify a negative colon cell population (HPA: Colon 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 carries no ICC-IF cell line for SEC61G; derive a cell-line control from the positive tissue's cell type (Glandular cells) and confirm it by RNA or western blot first.

Technical controls: Include a no-primary secondary-only control, a matched rabbit IgG isotype control, and an identically processed SEC61G knockout sample if available (A12389 tissue-IHC caption: rabbit primary antibody; standard IHC practice). Quench endogenous peroxidase and inspect inflammatory cells for background before interpreting colon HRP/DAB staining (A12389 tissue-IHC caption: HRP/DAB; standard IHC practice).

⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A12389 paraffin-section caption does not state a fixative (A12389 tissue-IHC caption: fixative not stated). The caption uses heat retrieval in EDTA at pH 8.0, but whether staining depends on retrieval is unreported (A12389 tissue-IHC caption: EDTA pH 8.0). Frozen sections or IF cannot be judged easier from the supplied evidence (HPA subcellular: no ICC-IF image-bearing cell lines); in colon IHC, endogenous peroxidase in inflammatory cells can produce misleading DAB signal (standard IHC practice).

HPA tissue IHC evidence for SEC61G

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Low consistency between antibody staining and RNA expression data. Pending external verification.). 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 →
Cerebellum	Purkinje cells	Medium	Protein (IHC)	HPA →
Colon	Glandular cells	Medium	Protein (IHC)	HPA →
Epididymis	Glandular cells	Medium	Protein (IHC)	HPA →
Esophagus	Squamous epithelial cells	Medium	Protein (IHC)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
Adipose tissue	Adipocytes	Not detected	Protein (IHC)	HPA →
Adrenal gland	Glandular cells	Not detected	Protein (IHC)	HPA →
Breast	Adipocytes	Not detected	Protein (IHC)	HPA →
Bronchus	Respiratory epithelial cells	Not detected	Protein (IHC)	HPA →
Caudate	Glial cells	Not detected	Protein (IHC)	HPA →

Section 3

Advanced SEC61G IHC Tips

These questions address SEC61G staining in chromogenic paraffin-section IHC, with one IF design question and attention to the limits of the available evidence.

Which retrieval conditions should I start with for SEC61G paraffin-section IHC?

Start with heat-mediated EDTA retrieval at pH 8.0 for paraffin sections (datasheet A12389). The selected tissue image pairs that retrieval with 2 μg/mL primary antibody overnight at 4°C, so keep those conditions stable while comparing retrieval runs (datasheet A12389). If staining remains weak, adjust heating duration within your validated instrument range before comparing another buffer, and process a matched control section in each run (standard IHC practice). Excessive retrieval can damage morphology and increase diffuse background, obscuring an ER-compatible cytoplasmic pattern (UniProt P60059 localisation; standard IHC practice).

How should I assess fixation when SEC61G staining varies between sections?

Target-specific fixation sensitivity is unknown, and the selected paraffin-section caption does not report its fixative (datasheet A12389). For new specimens, record fixative, fixation duration, tissue thickness, and processing, then compare matched sections under the same pH 8.0 EDTA retrieval conditions (datasheet A12389; standard IHC practice). Neither ER membrane topology nor reported tissue staining establishes which fixation time preserves this antibody's epitope (UniProt P60059 topology; HPA tissue IHC). Judge the preparation by retained morphology, reproducible cytoplasmic signal, and low no-primary background across replicate sections (standard IHC practice).

Where should SEC61G stain in paraffin sections, and which pattern needs investigation?

Expect cytoplasmic staining compatible with an ER membrane protein; isolated nuclear or exclusive cell-surface staining warrants investigation (UniProt P60059 localisation; HPA tissue IHC). SEC61G spans the membrane at residues 33–61, but chromogenic light microscopy cannot establish the orientation of its 1–32 and 62–68 termini (UniProt P60059 topology; standard IHC practice). Compare intact cells with a no-primary control and the section's morphology before accepting a perinuclear or granular pattern (standard IHC practice). HPA reports cytoplasmic expression in most tissues but rates its tissue-IHC reliability uncertain, so localisation alone cannot establish specificity (HPA tissue IHC).

Could an isoform or membrane-facing epitope explain weak SEC61G IHC staining?

SEC61G is 68 aa long, spans the membrane at 33–61, and has no annotated isoforms; absent staining alone does not establish an isoform change (UniProt P60059). Its 1–32 region is annotated as cytoplasmic and includes phosphoserine 18, while the opposite 62–68 tail is short (UniProt P60059 topology and modified residues). Those annotations do not identify the catalog antibody's epitope or establish a modification-dependent staining effect (UniProt P60059; datasheet A12389). Seek immunogen or epitope information, retain the documented pH 8.0 EDTA retrieval for comparisons, and use an independent specificity check when interpreting weak signal (datasheet A12389; standard IHC practice).

How should I assess SEC61G in multiplex IF alongside this IHC guide?

Use a marker for the expected cell type when assessing SEC61G colocalisation; colon glandular cells and lung macrophages are reported with medium tissue-IHC staining (HPA tissue IHC). Select fluorophores after checking the specimen's autofluorescence in each channel, particularly where broad background could resemble cytoplasmic signal (standard IF practice). Permeabilisation depends on epitope access: the 1–32 terminus is annotated cytoplasmic, whereas 62–68 is annotated on the opposite side of the ER membrane (UniProt P60059 topology). Confirm the antibody's epitope and assess access accordingly; its IHC conditions, including 2 μg/mL primary, do not establish IF performance (datasheet A12389; standard IF practice).

How can I reduce diffuse or patchy SEC61G DAB background?

First inspect no-primary and secondary-only sections to distinguish primary-associated staining from detection background (standard IHC practice). The selected image used 10% goat serum blocking, 2 μg/mL primary overnight at 4°C, and peroxidase detection with DAB; these are documented conditions for that image (datasheet A12389). Include an endogenous peroxidase block and check washing, secondary-antibody compatibility, and chromogen development time as general chromogenic IHC controls (standard IHC practice). Exclude tissue edges and damaged or necrotic regions when judging whether residual staining follows an intact-cell cytoplasmic pattern (UniProt P60059 localisation; standard IHC practice).

What scoring approach makes SEC61G IHC comparisons defensible? ⚠ ANSWER MARKED FOR VERIFICATION

Define the cell population and cytoplasmic compartment before scoring, since SEC61G is assigned to the ER membrane and HPA describes cytoplasmic tissue staining (UniProt P60059 localisation; HPA tissue IHC). Report the percentage of positive viable cells and an intensity score of 0–3; an H-score can combine intensity with each category's cell percentage on a 0–300 scale (standard IHC practice). Normalise counts to viable cells, or stained area to viable tissue area in mm², while excluding folds, edges, and necrosis (standard IHC practice). Keep retrieval, detection, exposure to DAB, and scoring thresholds consistent across sections; HPA rates its tissue-IHC reliability uncertain (datasheet A12389; HPA tissue IHC).

How can I distinguish a true SEC61G signal from chromogenic artefact?

Prioritise staining in intact-cell cytoplasm compatible with an ER membrane protein, then check whether the stained cell type matches the section's morphology (UniProt P60059 localisation; standard IHC practice). Medium staining is reported in colon glandular cells, but HPA rates SEC61G tissue-IHC reliability uncertain because antibody staining and RNA expression have low consistency (HPA tissue IHC). Investigate isolated nuclear staining, strong tissue-edge signal, necrotic deposits, and staining retained in a no-primary control before calling a section positive (UniProt P60059 localisation; standard IHC practice). An endogenous peroxidase block and matched detection controls help identify enzyme-derived DAB signal; confirm consequential findings with independent evidence (standard IHC practice).

Boster reagents

Best SEC61G / Protein transport protein Sec61 subunit gamma IHC Antibodies

A12389 has real IHC images from human cancer and mouse colon paraffin sections (A12389 image captions); its catalog lists human, mouse and rat reactivity, with no IF data (catalog: reactivity; IF image alts).

Real IHC data

Anti-SEC61G Antibody ®

Cat # A12389

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A12389 is listed for IHC and has images from paraffin sections of human colorectal adenocarcinoma, liver cancer, ovarian cancer and mouse colon (catalog: applications; A12389 image captions). It has no reported IF/ICC application or image (catalog: applications; IF image alts).

Which to pick: Choose A12389 for tissue IHC in paraffin sections; its caption uses EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody, but does not report the fixative (A12389 image captions). There is no catalog supported IF/ICC choice or reported clone for A12389 (catalog: applications; IF image alts; clone field). For cross species work, A12389 lists human, mouse and rat reactivity, while the supplied IHC images document human and mouse tissue only (catalog: reactivity; A12389 image captions).

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

References

UniProt Consortium. UniProt entry P60059 (SC61G_HUMAN, Protein transport protein Sec61 subunit gamma).
Human Protein Atlas. SEC61G tissue IHC expression (reliability: Uncertain).
Human Protein Atlas. SEC61G subcellular location (ICC-IF): Membrane.
Human Protein Atlas. SEC61G antibody validation summary (1 antibodies).
SEC61G Facilitates Brain Metastases via Antagonizing PGAM1 Ubiquitination and Immune Microenvironment Remodeling in Non-Small Cell Lung Cancer. International journal of biological sciences 2025 — PMC11844280.
SEC61 translocon gamma subunit is correlated with glycolytic activity, epithelial mesenchymal transition and the immune suppressive phenotype of lung adenocarcinoma. Acta biochimica et biophysica Sinica 2024 — PMC11693863.
Identification of SEC61G as a Diagnostic and Prognostic Biomarker in Oral Squamous Cell Carcinoma. Biomedicines 2023 — PMC10603851.
Machine learning-based prognostic model and single-cell transcriptomic integration for identifying brain metastasis-associated malignant subpopulations and potential therapeutic targets in lung adenocarcinoma. Cancer cell international 2026 — PMC13579860.
PubMed PMID:11042152 — UniProt-cited evidence.
PubMed PMID:14702039 — UniProt-cited evidence.
PubMed PMID:15489334 — UniProt-cited evidence.