RAGE Mouse mAb: FAQs, Applications & Protocols

RAGE Mouse mAb is a monoclonal antibody raised in mouse targeting the Receptor for Advanced Glycation End-products (RAGE), a multi-ligand receptor implicated in inflammation, diabetes complications, neurodegeneration, and cancer research.

Product SKU

Ab350673

Antibody Type

Mouse Monoclonal Antibody (mAb)

Formulation

PBS only (no carrier protein, no azide)

Target

RAGE (Receptor for Advanced Glycation End-products)

Host Species

Mouse

Reference / Cross-reference

absea#KC-2163

What Is RAGE and Why Is It an Important Research Target?

RAGE (Receptor for Advanced Glycation End-products) is a transmembrane receptor of the immunoglobulin superfamily encoded by the AGER gene. It binds a diverse range of ligands including advanced glycation end-products (AGEs), HMGB1, S100 proteins, and amyloid-β peptides. Upon ligand engagement, RAGE activates downstream signaling cascades—most notably NF-κB—driving pro-inflammatory gene expression.

RAGE is implicated in numerous disease contexts: diabetic vascular complications, Alzheimer's disease, atherosclerosis, sepsis, and tumor progression. Its expression is upregulated on endothelial cells, smooth muscle cells, neurons, and immune cells under pathological conditions, making it a valuable biomarker and therapeutic target. Researchers use validated anti-RAGE antibodies such as RAGE Mouse mAb (Ab350673) to detect, localize, and quantify RAGE expression in cellular and tissue models.

Recommended Applications and Suggested Working Dilutions

RAGE Mouse mAb (Ab350673) is designed for use in standard immunological detection applications. Typical applications for anti-RAGE monoclonal antibodies of this class include:

• Western Blot (WB): Detects RAGE protein in cell or tissue lysates under denaturing conditions. Optimization of primary antibody dilution is recommended for each laboratory's specific lysate and detection system.
• Immunohistochemistry (IHC): Localizes RAGE expression in formalin-fixed paraffin-embedded (FFPE) or frozen tissue sections. Antigen retrieval may be required for FFPE sections.
• Immunofluorescence (IF): Enables visualization of RAGE subcellular localization in fixed cell preparations.
• ELISA: May be used as a capture or detection antibody in sandwich ELISA formats for quantitative RAGE measurement.

Note: Specific validated dilution ranges and confirmed reactive species are not detailed in the current source data. Users should consult the product datasheet for Ab350673 or contact Aladdin Scientific technical support for up-to-date application notes and titration recommendations.

Storage, Handling, and Stability

RAGE Mouse mAb (Ab350673) is supplied in PBS only (phosphate-buffered saline, without carrier protein or preservative). This formulation is suitable for direct conjugation and avoids interference from BSA or sodium azide in downstream assays.

General storage guidelines for PBS-formulated monoclonal antibodies:

• Store at –20 °C for long-term storage. Avoid repeated freeze-thaw cycles, which can lead to aggregation and loss of activity.
• For short-term use (up to 1 month), the antibody may be stored at 4 °C.
• Aliquot into single-use volumes upon first thaw to minimize freeze-thaw cycling.
• Keep away from light and contamination. Do not store in frost-free freezers that undergo automatic defrost cycles.

Important: Always verify storage conditions on the official Ab350673 product datasheet, as lot-specific recommendations may apply.

Basic Protocol for Western Blot Using RAGE Mouse mAb

The following is a general-purpose WB protocol framework for use with RAGE Mouse mAb (Ab350673). Optimization for specific cell/tissue types and detection systems is required.

1. Sample preparation: Lyse cells or homogenize tissue in RIPA buffer supplemented with protease inhibitors. Determine protein concentration (e.g., BCA assay). Load 20–40 µg total protein per lane.
2. SDS-PAGE: Resolve proteins on an appropriate percentage polyacrylamide gel (typically 10–12% for RAGE, ~45–50 kDa). Run under reducing conditions unless otherwise specified.
3. Transfer: Transfer resolved proteins to PVDF or nitrocellulose membrane using wet or semi-dry transfer.
4. Blocking: Block membrane for 1 hour at room temperature in 5% non-fat dry milk or 3–5% BSA in TBST.
5. Primary antibody incubation: Incubate with RAGE Mouse mAb (Ab350673) at the recommended dilution (see datasheet) in blocking buffer overnight at 4 °C or 1–2 hours at room temperature.
6. Washing: Wash 3 × 10 minutes with TBST.
7. Secondary antibody: Incubate with an HRP-conjugated anti-mouse IgG secondary antibody at manufacturer-recommended dilution for 1 hour at room temperature.
8. Detection: Develop using ECL substrate and image using a chemiluminescence detection system.

Note: Specific dilution ratios for Ab350673 are not confirmed in the source data and should be obtained from the product datasheet or Aladdin Scientific technical support.

Troubleshooting Common Issues with RAGE Mouse mAb

Below are common troubleshooting scenarios when working with RAGE Mouse mAb (Ab350673):

Problem	Possible Cause	Suggested Action
No signal / faint band	Antibody concentration too low; insufficient antigen; poor transfer	Increase antibody concentration; use positive control lysate; verify transfer efficiency with Ponceau S staining
High background	Insufficient blocking; antibody concentration too high; non-specific binding	Increase blocking time/concentration; optimize antibody dilution; increase wash stringency
Multiple non-specific bands	Protein degradation; cross-reactivity	Use fresh samples with protease inhibitors; confirm expected molecular weight (~45–50 kDa for RAGE)
Loss of activity after storage	Repeated freeze-thaw; improper storage temperature	Aliquot antibody; store at –20 °C; avoid frost-free freezers

Contact Aladdin Scientific technical support for application-specific troubleshooting assistance.

Frequently asked questions

What is the formulation of RAGE Mouse mAb (Ab350673)?

RAGE Mouse mAb (Ab350673) is formulated in PBS only, without carrier proteins such as BSA or preservatives such as sodium azide. This makes it suitable for direct labeling or conjugation applications where such additives could interfere.

Which species does RAGE Mouse mAb (Ab350673) react with?

The confirmed reactive species for Ab350673 are not specified in the current source material. Please consult the official product datasheet available on the Aladdin Scientific website or contact technical support for verified species reactivity data.

How should I store RAGE Mouse mAb (Ab350673) to maintain activity?

For long-term storage, keep the antibody at –20 °C. For short-term use, storage at 4 °C for up to one month is generally acceptable. Aliquot the antibody into single-use volumes upon first thaw to avoid repeated freeze-thaw cycles, which can cause aggregation and reduce performance.

What applications is RAGE Mouse mAb (Ab350673) suitable for?

RAGE Mouse mAb is generally suitable for immunological applications including Western Blot, Immunohistochemistry, Immunofluorescence, and ELISA. However, specific validated applications for Ab350673 should be confirmed on the product datasheet, as not all applications have been verified for every antibody lot.

What is the cross-reference number for Ab350673?

Ab350673 (RAGE Mouse mAb) cross-references to absea catalog number KC-2163. This reference can be used to locate additional technical documentation or equivalent product information from the originating supplier.

What is the expected molecular weight of RAGE detected by this antibody?

The RAGE protein (encoded by the AGER gene) has a theoretical molecular weight of approximately 42–50 kDa, though the observed band on Western Blot may appear slightly higher due to glycosylation. Please verify the expected band size against published literature and the product datasheet.

References

1. Aladdin Scientific. Ab350673 – RAGE Mouse mAb product page. Aladdin Scientific.
2. Bierhaus A, Humpert PM, Morcos M, et al. Understanding RAGE, the receptor for advanced glycation end products. J Mol Med. 2005;83(11):876-886.
3. Schmidt AM, Yan SD, Yan SF, Stern DM. The biology of the receptor for advanced glycation end products and its ligands. Biochim Biophys Acta. 2000;1498(2-3):99-111.

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