IGSF3 Mouse mAb: Selection, Application & Protocol Guide

IGSF3 Mouse mAb is a mouse-derived monoclonal antibody raised against human IGSF3 protein, designed to deliver high specificity and reproducibility across key immunoassay applications including western blot, immunohistochemistry, and immunocytochemistry.

Product SKU

Ab351255

Clone Origin

Mouse monoclonal

Formulation

PBS only (no BSA, no glycerol, no azide unless specified)

Target Protein

IGSF3 (Immunoglobulin Superfamily Member 3)

Recommended Storage

Store at -20 °C; avoid repeated freeze-thaw cycles

Species Reactivity

Human (verify cross-reactivity with supplier before use)

Target Background: IGSF3 Biology

IGSF3 (Immunoglobulin Superfamily Member 3) is a single-pass type I transmembrane protein belonging to the immunoglobulin superfamily. It contains multiple immunoglobulin-like C2-type domains and is involved in cell–cell adhesion, signal transduction at the cell surface, and potentially in immune-cell interactions. IGSF3 is expressed in a variety of tissues and has been implicated in cancer biology, where altered expression may influence tumor cell behavior and patient prognosis.

Because IGSF3 resides at the plasma membrane and participates in extracellular protein–protein interactions, antibody-based detection methods such as immunohistochemistry (IHC) and immunofluorescence (IF) are particularly well suited to studying its expression patterns in tissues and cultured cells.

Antibody Specifications & Formulation

The IGSF3 Mouse mAb (Ab351255) is a mouse monoclonal antibody supplied in PBS-only formulation. The PBS-only buffer system offers flexibility for researchers who need to prepare custom conjugates, add their own stabilizers, or perform direct labeling without interference from additives such as BSA or sodium azide.

• Format: Purified mouse monoclonal IgG in PBS
• Preservative: No azide added (confirm with lot-specific datasheet)
• Carrier protein: No BSA added (confirm with lot-specific datasheet)
• Concentration: Refer to the vial label and certificate of analysis for the lot-specific concentration

Always consult the lot-specific Certificate of Analysis (CoA) provided by Aladdin Scientific for confirmed concentration, purity, and endotoxin data before use.

Recommended Protocol: Western Blot (WB)

The following protocol provides a starting framework for western blot detection of IGSF3. Optimization of antibody dilution and incubation conditions may be required depending on the sample type and detection system used.

1. Sample preparation: Lyse cells or homogenize tissue in RIPA buffer supplemented with protease inhibitor cocktail. Determine protein concentration by BCA or Bradford assay.
2. Gel electrophoresis: Load 20–40 µg of total protein per lane on a 10–12% SDS-PAGE gel under reducing conditions.
3. Transfer: Transfer proteins to PVDF or nitrocellulose membrane at 100 V for 60–90 min (wet transfer) or according to your system's optimized settings.
4. Blocking: Block membrane with 5% non-fat dry milk or 5% BSA in TBST for 1 h at room temperature.
5. Primary antibody incubation: Dilute IGSF3 Mouse mAb (Ab351255) in blocking buffer (suggested starting dilution 1:500–1:2000; optimize empirically). Incubate overnight at 4 °C with gentle rocking.
6. Washing: Wash 3× with TBST, 10 min each.
7. Secondary antibody: Incubate with an HRP-conjugated anti-mouse IgG secondary antibody (1:5000–1:10000) for 1 h at room temperature.
8. Detection: Develop with ECL substrate and image using X-ray film or a digital imaging system.

Note: Specific dilution ranges and expected band sizes have not been confirmed in the source material and should be validated against the lot-specific datasheet.

Storage, Handling & Quality Control

Proper storage and handling of monoclonal antibodies are critical to maintaining binding activity and batch-to-batch consistency.

• Long-term storage: Store at -20 °C in aliquots to avoid repeated freeze-thaw cycles. Single-use aliquots are strongly recommended.
• Short-term storage: If the antibody will be used within 1–2 weeks, it may be stored at 4 °C. Do not store at 4 °C for extended periods.
• Avoid: Repeated freeze-thaw cycles (>3 cycles may reduce activity), exposure to direct light, and high temperatures.
• PBS-only formulation note: Because this antibody is supplied in PBS without stabilizers such as glycerol or BSA, particular care should be taken during freeze-thaw to maintain protein integrity. Consider adding 50% glycerol (v/v) for long-term storage at -20 °C if recommended by the lot-specific datasheet.
• Quality control: Each lot is tested by Aladdin Scientific prior to release. Verify specifics (purity, concentration, functional test results) on the CoA included with each shipment.

Troubleshooting Common Issues

The following guidance addresses frequently encountered issues when working with IGSF3 Mouse mAb.

Problem	Likely Cause	Suggested Action
No signal detected	Antibody dilution too high; insufficient antigen; inadequate transfer	Reduce dilution; use positive control lysate; verify transfer efficiency with Ponceau S staining
High background	Insufficient blocking; secondary antibody cross-reactivity; primary antibody concentration too high	Increase blocking time or concentration; increase primary antibody dilution; optimize wash steps
Multiple non-specific bands	Protein degradation; cross-reactivity	Add fresh protease inhibitors; confirm sample integrity by Coomassie staining; validate with knockout or knockdown control
Weak signal in IHC	Inadequate antigen retrieval; low antibody concentration	Optimize antigen retrieval (heat-induced vs. enzymatic); increase antibody concentration or incubation time

Frequently asked questions

What is the recommended starting dilution for IGSF3 Mouse mAb in western blot?

A starting dilution of 1:500 to 1:2000 is commonly used for mouse monoclonal antibodies in western blot; however, the optimal dilution for Ab351255 should be determined empirically using a positive control sample. Always refer to the lot-specific datasheet provided by Aladdin Scientific for validated conditions.

Why is IGSF3 Mouse mAb supplied in PBS only, and does this affect storage?

PBS-only formulation gives researchers maximum flexibility for downstream conjugation, custom stabilizer addition, or direct labeling without interference from carrier proteins or preservatives. Because there is no glycerol or azide, the antibody is more susceptible to freeze-thaw damage; aliquoting before freezing and limiting freeze-thaw cycles to fewer than three is strongly advised.

Is IGSF3 Mouse mAb suitable for immunohistochemistry on FFPE sections?

Mouse monoclonal antibodies are generally compatible with FFPE immunohistochemistry when appropriate antigen retrieval is performed. Optimization of retrieval method (heat-induced epitope retrieval with citrate buffer pH 6.0 or EDTA buffer pH 9.0) and antibody dilution is recommended. Specific validation on FFPE tissue has not been confirmed in the available source material; please contact Aladdin Scientific or check the product datasheet for application-specific validation data.

What positive control should I use when validating IGSF3 Mouse mAb?

Cell lines or tissues known to express IGSF3 at moderate-to-high levels should be used as positive controls. A negative control using an isotype-matched mouse IgG at the same concentration is essential to assess non-specific binding. Knockout or siRNA knockdown cell lines, if available, provide the most stringent specificity confirmation.

Can IGSF3 Mouse mAb be used for immunoprecipitation (IP)?

Suitability for immunoprecipitation depends on whether the antibody recognizes the native conformation of IGSF3 under non-denaturing lysis conditions. This specific application has not been confirmed in the available source material for Ab351255. Contact Aladdin Scientific technical support for guidance or request application-specific validation data.

References

1. Aladdin Scientific product page: IGSF3 Mouse mAb, Ab351255, PBS formulation (internal reference KC-31036).
2. Bhatt DL et al. Immunoglobulin superfamily members in cell adhesion and cancer — general textbook-level reference; specific IGSF3 citations not confirmed in source material.

Products are supplied for research and development use only. Not for use in humans, animals, diagnosis, or therapy.