CISD1 / CDGSH iron-sulfur domain-containing protein 1 · IHC design guide

Design Immunohistochemistry for CISD1

Plan chromogenic paraffin IHC for CISD1 with the IHC-validated antibody (datasheet A04360-2). Compare granular cytoplasmic staining with high staining in colon glandular cells, while accounting for medium consistency between staining and RNA expression (HPA tissue IHC).

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

Immunohistochemistry protocol sheet for CISD1 (IHC for CISD1): expected localisation Granular cytoplasm (HPA tissue IHC); mitochondrial outer membrane (UniProt), antibody A04360-2, validated IHC image, and IHC protocol steps — Printable **CISD1** IHC protocol sheet — expected localisation Granular cytoplasm (HPA tissue IHC); mitochondrial outer membrane (UniProt), antibody A04360-2, controls and protocol steps. Open the full CISD1 IHC guide →

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CISD1 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	Granular cytoplasm (HPA tissue IHC); mitochondrial outer membrane (UniProt)
	Staining pattern	Granular cytoplasm in most tissues; high in colon glands (HPA tissue IHC)
	Antigen retrieval	EDTA pH 8.0 HIER, heat-mediated (datasheet A04360-2)
	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); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A04360-2)
	Caveat	Staining has medium consistency with RNA expression (HPA tissue IHC)
	Regulation	Reduced in cystic fibrosis cells (UniProt)
	Isoform / epitope	No annotated isoforms; residues 32–108 face cytoplasm (UniProt)

Section 1

Recommended CISD1 IHC & IF Protocols

The catalog antibody protocol is paired with published CISD1 IHC methods for breast, lung, and pancreatic cancer sections (PMC9729735; PMC11948984; PMC8760727).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Paraffin-embedded human gastric cancer tissue; fixative not specified (datasheet A04360-2)
Fixation	Image fixative and duration unreported (datasheet A04360-2); 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 A04360-2); 20 min, 95–100 °C (standard)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% goat serum (datasheet A04360-2)
Primary antibody	Rabbit anti-CISD1, 2-5μg/ml (datasheet A04360-2)
Primary incubation	Overnight at 4 °C (datasheet A04360-2)
Detection	Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A04360-2)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	CISD1-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: Granular cytoplasmic expression in most tissues. No signal in the no-primary control.

💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet A04360-2). For replication, follow each article’s stated retrieval conditions (PMC11948984; PMC8760727).

Section 2

What Is the Expected CISD1 Staining Pattern?

CISD1 is an outer mitochondrial membrane protein with a cytoplasmic domain (UniProt Q9NZ45 topology). In paraffin-section IHC, expect granular cytoplasmic staining in many tissues, including colon glandular cells and heart cardiomyocytes (HPA tissue IHC). HPA rates its tissue pattern Supported, with medium agreement between staining and RNA data; external verification is pending (HPA tissue IHC).

What am I looking at on my slide?
Granular cytoplasmic staining in colon or appendix glandular cells, with little signal in adjacent unstained areas.	This matches the reported pattern and High staining in those cell types (HPA tissue IHC). Score the named cells and their intensity rather than treating an entire tissue section as uniformly positive. Chromogenic IHC alone cannot prove that each granule is a mitochondrion.
Predominantly nuclear or sharply plasma-membrane staining replaces the granular cytoplasmic pattern.	That distribution conflicts with the reported tissue pattern (HPA tissue IHC) and mitochondrial outer-membrane location (UniProt Q9NZ45). Consider nonspecific staining or a detection artefact; review controls before assigning a biological interpretation.
Strong staining in lung alveolar cells or adipocytes.	Those specific cell populations were Not detected in the HPA tissue survey (HPA tissue IHC). Check for antibody cross-reactivity and endogenous detection activity. An HPA negative observation is a comparison point, not proof that every specimen of that cell type must be negative.
Uniform brown haze covers cells, stroma and empty areas without a discernible granular cell pattern.	The distribution does not resemble the reported granular cytoplasmic staining (HPA tissue IHC). General IHC practice: compare a no-primary control and inspect washing, blocking and chromogen development before scoring the haze as CISD1.
No staining appears in colon glandular cells or heart cardiomyocytes while the run otherwise looks interpretable.	Both are reported High-staining cell types (HPA tissue IHC), so an absent signal warrants a technical check. Because HPA rates the tissue evidence Supported and pending external verification (HPA tissue IHC), avoid declaring a biological loss from one section alone.

💡Expected CISD1 appearanceA positive IHC result is granular cytoplasmic staining in expected cells, potentially High in colon glandular cells or cardiomyocytes (HPA tissue IHC); nuclear staining or uniform brown haze is discordant with that pattern (HPA tissue IHC; UniProt Q9NZ45 location).

How each factor affects the staining
Compartment and topology	CISD1 has a membrane segment at 14–31 and a cytoplasmic region at 32–108 (UniProt Q9NZ45 topology). Use the reported granular cytoplasmic IHC pattern for slide interpretation (HPA tissue IHC); light microscopy does not resolve membrane orientation.
Choice of comparison cells	Colon glandular cells and heart cardiomyocytes are High, while lung alveolar cells and adipocytes are Not detected in the surveyed sections (HPA tissue IHC). Compare the specified cell populations, not whole-tissue labels.
Strength of tissue evidence	The HPA tissue pattern is Supported, with medium staining–RNA agreement and external verification pending (HPA tissue IHC). Its antibody HPA074383 has Supported IHC validation (HPA antibodies); this does not establish specificity in every specimen.
IF/ICC Q: What localisation should an independent image show?	Mitochondrial localisation is Supported in HPA ICC-IF images from PC-3 and U2OS cells (HPA subcellular). Treat that as a compartment crosscheck; the IF/ICC guide covers its own workflow.

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
Expected glandular cells are blank.	Colon and appendix glandular cells are reported High (HPA tissue IHC), so a blank result may reflect a failed IHC run or a specimen difference.	General IHC practice: examine a same-run positive control, section integrity, retrieval and primary-antibody application; repeat with the IHC-validated antibody's documented conditions before interpreting absence.
Signal appears mainly in nuclei.	Nuclear dominance is discordant with mitochondrial location (UniProt Q9NZ45) and the granular cytoplasmic IHC profile (HPA tissue IHC).	General IHC practice: check the no-primary control, counterstain and detection reagents; review antibody concentration and confirm that any retained signal follows the expected cell pattern.
Lung alveolar cells stain strongly.	These cells were Not detected in the HPA survey (HPA tissue IHC); cross-reactivity or endogenous detection activity is possible.	General IHC practice: compare a no-primary control, check endogenous-enzyme blocking for chromogenic detection and assess a same-run expected-positive cell population before scoring the result.
Brown haze is present across the section.	A uniform deposit lacks the reported granular cytoplasmic distribution (HPA tissue IHC) and may arise from background in the staining workflow.	General IHC practice: inspect the no-primary control, washing and reagent coverage; adjust blocking or detection development according to the validated workflow, then reassess cellular detail.
Some fields stain while comparable cells in other fields do not.	The uneven distribution may reflect section or reagent-coverage variation; HPA's cell-specific intensity calls do not establish a fixation effect (HPA tissue IHC).	General IHC practice: inspect section integrity and reagent coverage, compare matched cell types across fields and repeat the section if technical variation prevents reliable scoring.
The no-primary control develops brown signal.	General IHC practice: signal without primary antibody points to the detection system or endogenous activity, so it cannot be assigned to CISD1.	General IHC practice: resolve the control signal using appropriate endogenous-activity blocking and detection controls, then repeat the IHC run before interpreting CISD1 staining.

Source: UniProt Q9NZ45 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing CISD1 staining

IHC analysis of MitoNEET/CISD1 using anti-MitoNEET/CISD1 antibody (A04360-2). MitoNEET/CISD1 was detected in paraffin-embedded section of human gastric cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-MitoNEET/CISD1 Antibody (A04360-2) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

A04360-2 · IHCIHC analysis of MitoNEET/CISD1 using anti-MitoNEET/CISD1 antibody (A04360-2). MitoNEET/CISD1 was detected in paraffin-embedded section of hu

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000122873-CISD1/tissue/cerebral+cortex#imid_20849319

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000122873-CISD1/tissue/colon#imid_20849245

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000122873-CISD1/tissue/liver#imid_20849246

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000122873-CISD1/tissue/kidney#imid_20849247

HPA · ICC-IF PC-3Human Protein Atlas immunofluorescence, PC-3, antibody HPA074383; green = CISD1, blue = nucleus, red = microtubules.

← swipe / scroll for more images →

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

Sample controls for CISD1 IHC & IF

🧪Run colon first: glandular cells should stain (HPA: High in colon glandular cells); use cervix glandular cells as a negative tissue comparator (HPA: Not detected in cervix glandular cells). Within the colon section, cells without specific staining should remain at background, but the supplied HPA rows do not identify a defined internal negative cell population (HPA: colon glandular cells High).

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 CISD1 in PC-3, U2OS, with annotated localisation: Mitochondria (supported) (HPA subcellular).

Technical controls: Include a no-primary, secondary-only control; a concentration-matched rabbit IgG isotype control for the catalog antibody (selected-SKU IHC caption: rabbit primary); and a CISD1 knockout specimen or peptide-blocked primary, if available. Quench endogenous peroxidase and check for endogenous biotin staining in the colon section because the reported detection uses a biotinylated secondary, streptavidin–biotin complex, and DAB (selected-SKU IHC caption).

⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU paraffin-section caption does not state the fixative (selected-SKU IHC caption: fixative not stated). The reported IHC procedure uses heat retrieval in EDTA at pH 8.0, but the payload does not establish whether retrieval is required (selected-SKU IHC caption). A frozen-section or IF advantage is unreported; for this IHC tissue, inspect glandular background from the biotin-based detection system, while mitochondrial ICC-IF localisation is separately supported in PC-3 and U2OS cells (selected-SKU IHC caption; HPA: mitochondria supported, PC-3 and U2OS ICC-IF images).

HPA tissue IHC evidence for CISD1

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium 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	High	Protein (IHC)	HPA →
Cerebellum	Cells in molecular layer	High	Protein (IHC)	HPA →
Cerebral cortex	Neuropil	High	Protein (IHC)	HPA →
Colon	Glandular cells	High	Protein (IHC)	HPA →
Duodenum	Glandular cells	High	Protein (IHC)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
Adipose tissue	Adipocytes	Not detected	Protein (IHC)	HPA →
Bone marrow	Hematopoietic cells	Not detected	Protein (IHC)	HPA →
Caudate	Glial cells	Not detected	Protein (IHC)	HPA →
Cervix	Glandular cells	Not detected	Protein (IHC)	HPA →
Lung	Alveolar cells	Not detected	Protein (IHC)	HPA →

Section 3

Advanced CISD1 IHC Tips

Troubleshoot chromogenic CISD1 staining in paraffin sections using the catalog antibody’s tissue image, protein topology, and reported tissue staining patterns.

What retrieval should I start with for CISD1 in paraffin sections?

Start with heat-mediated EDTA retrieval at pH 8.0 for paraffin sections (datasheet A04360-2). This is the retrieval used for the catalog antibody’s human gastric cancer tissue image, followed by 2 µg/mL primary antibody overnight at 4°C (datasheet A04360-2). If staining is weak, first check section adhesion, heating consistency, and whether the positive control stains in the same run (standard IHC practice). Compare any adjusted retrieval condition on serial sections while holding antibody concentration and detection constant, because retrieval changes can also raise background or damage tissue morphology (standard IHC practice).

Could fixation explain weak or uneven CISD1 staining?

Target-specific sensitivity of CISD1 to fixation is unknown from the supplied evidence; the catalog tissue caption identifies a paraffin section but does not state its fixative (datasheet A04360-2). Record the fixative and processing history for each specimen, then compare affected sections with a known staining control processed alongside them (standard IHC practice). Overfixation, underfixation, and uneven processing can alter antigen accessibility or morphology in IHC generally, but their effects on CISD1 have not been established here (standard IHC practice). If staining differs between batches, hold retrieval, 2 µg/mL primary concentration, and detection constant while reviewing the processing records (datasheet A04360-2; standard IHC practice).

Where should convincing CISD1 chromogenic staining appear?

Expect a granular cytoplasmic pattern rather than a predominantly nuclear or plasma-membrane signal (HPA: granular cytoplasmic expression in most tissues; UniProt Q9NZ45: mitochondrial outer membrane). CISD1 has a transmembrane segment at residues 14–31 and a cytoplasmic region at 32–108, so light microscopy should be interpreted at the organelle-pattern level rather than as resolved membrane topology (UniProt Q9NZ45 topology; standard IHC practice). The HPA reports high staining in colon glandular cells and heart cardiomyocytes, providing tissue and cell-context references (HPA tissue IHC). Examine matched negative-primary controls and compare staining within intact cells, especially where dense cytoplasm or pigment could mimic granules (standard IHC practice).

How do topology and epitope uncertainty affect antibody troubleshooting?

The supplied UniProt record lists 0 isoforms and places the cytoplasmic portion of CISD1 at residues 32–108 (UniProt Q9NZ45 isoforms and topology). It also lists acetylation at residues 2, 55, 68, and 104, but these annotations alone do not establish an effect on tissue staining (UniProt Q9NZ45 modified residues). An antibody epitope is not specified in the supplied catalog image, so do not assume which side of the membrane it recognizes (datasheet A04360-2). If results disagree across reagents, obtain each antibody’s epitope information and compare serial-section staining with the same retrieval and detection conditions (standard IHC practice).

How can IF help investigate a questionable CISD1 IHC pattern?

Use IF as a complementary localisation check: multiplex CISD1 with a mitochondrial marker and, in colon sections, a marker identifying glandular cells (HPA subcellular: mitochondria; HPA tissue IHC: high in colon glandular cells; standard IF practice). Select spectrally separated fluorophores, favoring a far-red channel when tissue autofluorescence obscures shorter wavelengths, and inspect single-label controls (standard IF practice). Because residues 32–108 face the cytoplasm, permeabilisation may be needed if the antibody recognizes an epitope there; its epitope is unspecified, so optimize access empirically (UniProt Q9NZ45 topology; datasheet A04360-2; standard IF practice). Interpret agreement with IHC by cellular compartment and cell type, while accounting for the different detection methods (standard IHC/IF practice).

How should I investigate diffuse DAB or tissue-wide background?

The catalog tissue image used 10% goat-serum block, biotinylated goat anti-rabbit secondary antibody, a streptavidin–biotin complex, and DAB (datasheet A04360-2). Run no-primary and secondary-only controls to separate nonspecific secondary or detection signal from primary-associated staining (standard IHC practice). With a biotin-based detection system, assess endogenous biotin; also include a peroxidase block and inspect endogenous pigment before attributing brown deposits to CISD1 (datasheet A04360-2; standard IHC practice). If background remains high, compare controls before changing primary concentration, and judge any improvement against the expected granular cytoplasmic pattern in intact cells (HPA tissue IHC; standard IHC practice).

How should I score CISD1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION

Define the cell population and tissue region before scoring, then record both the percentage of positive cells and cytoplasmic staining intensity (standard IHC practice). An H-score can combine intensity categories with their cell percentages on a 0–300 scale, provided the same thresholds and scoring rules are applied to every section (standard IHC practice). For spatial analysis, report positive cells per mm² of viable tissue and exclude necrotic or folded areas using prespecified rules (standard IHC practice). Normalize comparisons to the same eligible cell population, sampled area, and staining batch, and retain a common control section to monitor run-to-run variation (standard IHC practice).

What distinguishes true CISD1 staining from artefact?

A plausible positive signal is granular and cytoplasmic in intact cells, consistent with the reported tissue profile and mitochondrial location (HPA tissue IHC; HPA subcellular). Check that the stained cell type fits the specimen: colon glandular cells are reported as high, whereas lung alveolar cells are reported as not detected (HPA tissue IHC). Predominantly nuclear staining, staining confined to section edges or necrotic areas, and signal reproduced in no-primary controls warrant investigation before assigning CISD1 positivity (UniProt Q9NZ45 localisation; standard IHC practice). Exclude pigment and endogenous peroxidase or biotin contributions, particularly with the catalog image’s biotin-based DAB detection workflow (datasheet A04360-2; standard IHC practice).

Boster reagents

Best CISD1 / CDGSH iron-sulfur domain-containing protein 1 IHC Antibodies

A04360-2 has IHC data from paraffin-embedded human gastric cancer tissue (catalog IHC image caption) and lists Human, Mouse, Rat reactivity (catalog reactivity). No IF data are supplied (catalog IF image alts).

Real IHC data

Anti-MitoNEET/CISD1 Antibody ®

Cat # A04360-2

View product →

A04360-2 is the only SKU that will render (catalog payload). Its IHC image shows staining in a paraffin-embedded human gastric cancer section; the fixative is unreported (catalog IHC image caption).

Which to pick: Choose A04360-2 for paraffin-section IHC using a rabbit antibody (catalog IHC image caption; catalog host). No listed SKU has IF/ICC validation (catalog applications; catalog IF image alts). A04360-2 lists Human, Mouse, Rat reactivity, but its illustrated IHC result is from human tissue only (catalog reactivity; catalog IHC image caption).

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

References

UniProt Consortium. UniProt entry Q9NZ45 (CISD1_HUMAN, CDGSH iron-sulfur domain-containing protein 1).
Human Protein Atlas. CISD1 tissue IHC expression (reliability: Supported).
Human Protein Atlas. CISD1 subcellular location (ICC-IF): Localized to the mitochondria..
Human Protein Atlas. CISD1 antibody validation summary (1 antibodies).
Identification of CISD1 as a Prognostic Biomarker for Breast Cancer. International journal of general medicine 2022 — PMC9729735.
Predictive and therapeutic value of the ferroptosis gene CISD1 in non?small cell lung cancer. Oncology letters 2025 — PMC11948984.
Systematic Analysis and Validation of the Prognosis, Immunological Role and Biology Function of the Ferroptosis-Related lncRNA GSEC/miRNA-101-3p/CISD1 Axis in Lung Adenocarcinoma. Frontiers in molecular biosciences 2021 — PMC8936422.
Development and validation of a novel 3-gene prognostic model for pancreatic adenocarcinoma based on ferroptosis-related genes. Cancer cell international 2022 — PMC8760727.
PubMed PMID:18047834 — UniProt-cited evidence.
PubMed PMID:14702039 — UniProt-cited evidence.
PubMed PMID:15489334 — UniProt-cited evidence.