MAP3K5 Mouse mAb: Usage Guidelines & Technical Overview

MAP3K5 Mouse mAb is a monoclonal antibody targeting MAP3K5 (also known as ASK1), formulated in PBS and designed to support reproducible detection of this stress-activated kinase in a range of research applications.

Product SKU

Ab351000

Antibody Format

Mouse Monoclonal Antibody (mAb)

Formulation

PBS (Phosphate-Buffered Saline)

Target Protein

MAP3K5 (Mitogen-Activated Protein Kinase Kinase Kinase 5, also known as ASK1)

Host Species

Mouse

Reference Source

absea#KC-1340

Product Overview and Target Biology

MAP3K5 Mouse mAb (Ab351000) is a mouse-derived monoclonal antibody directed against MAP3K5, also commonly referred to as Apoptosis Signal-regulating Kinase 1 (ASK1). MAP3K5 is a serine/threonine kinase and a member of the mitogen-activated protein kinase kinase kinase (MAP3K) family. It plays a central role in stress-induced signaling cascades, activating downstream MKK3/MKK6–p38 and MKK4/MKK7–JNK pathways in response to oxidative stress, endoplasmic reticulum stress, and inflammatory cytokines.

Due to its pivotal involvement in apoptosis, inflammation, and cellular stress responses, MAP3K5/ASK1 is of broad scientific interest in fields including oncology, cardiovascular research, neurodegenerative disease, and immunology. This monoclonal antibody provides a highly specific tool for researchers seeking to detect and study endogenous MAP3K5 protein expression and activation status.

Formulation, Storage, and Handling

MAP3K5 Mouse mAb (Ab351000) is supplied in Phosphate-Buffered Saline (PBS) without additional carrier proteins or preservatives, unless otherwise specified on the Certificate of Analysis. PBS-only formulation is advantageous when researchers require direct conjugation, custom formulation, or assays sensitive to carrier proteins such as BSA or sodium azide.

Recommended Storage Conditions:

• Store at –20 °C for long-term storage. Avoid repeated freeze–thaw cycles, as these can reduce antibody activity and specificity.
• For short-term use (up to 1–2 weeks), aliquots may be stored at 4 °C.
• Prepare single-use aliquots upon first thaw to minimize freeze–thaw degradation.

Handling Tips:

• Centrifuge the vial briefly before opening to collect contents at the bottom.
• Use low-protein-binding tubes and pipette tips to minimize antibody loss.
• Avoid prolonged exposure to room temperature or direct light.

Note: Specific lot concentration and exact volume should be confirmed on the product datasheet or Certificate of Analysis. Please contact Aladdin Scientific for lot-specific information.

Recommended Application Protocols

MAP3K5 Mouse mAb is suitable for use in standard immunological and biochemical research applications. Although specific validated applications should be confirmed on the product datasheet, monoclonal antibodies of this type are commonly employed in the following assays:

• Western Blotting (WB): Denature and separate cell lysates by SDS-PAGE; transfer to PVDF or nitrocellulose membrane; block with 5% non-fat milk or BSA in TBST; incubate with primary antibody at recommended dilution (to be confirmed per datasheet) overnight at 4 °C; apply HRP-conjugated anti-mouse secondary antibody; detect by chemiluminescence.
• Immunoprecipitation (IP): Pre-clear cell lysate; add MAP3K5 Mouse mAb and incubate with rotation at 4 °C; capture with Protein A/G beads; wash thoroughly; elute and analyze by Western blot.
• Immunofluorescence (IF): Fix and permeabilize cells; block with normal serum; incubate with primary antibody; apply fluorophore-conjugated anti-mouse secondary antibody; counterstain nuclei; image by fluorescence microscopy.

General Recommendations: Optimize antibody dilution, blocking conditions, and incubation times for each specific application and cell/tissue type. Include appropriate positive and negative controls in every experiment.

Note: Specific validated dilutions and confirmed application list must be verified against the official product datasheet (Ab351000). The above applications represent general guidance for antibodies of this class.

Quality Control and Troubleshooting

Consistent results with MAP3K5 Mouse mAb depend on rigorous quality control practices and systematic troubleshooting when unexpected results arise.

Quality Control Practices:

• Always include a positive control lysate or cell line known to express MAP3K5 (e.g., human cancer cell lines such as HeLa or HEK293 that endogenously express ASK1).
• Use a negative control (primary antibody omission or isotype control mouse IgG) to assess background and non-specific binding.
• Confirm antibody integrity by checking lot-specific Certificate of Analysis documentation provided by Aladdin Scientific.

Common Troubleshooting Scenarios:

• No signal or weak signal: Increase antibody concentration or extend primary antibody incubation time; verify that secondary antibody is anti-mouse; confirm cell lysis efficiency and protein loading.
• High background: Increase blocking time or concentration; optimize wash steps; reduce primary antibody concentration.
• Multiple non-specific bands: Optimize SDS-PAGE conditions; verify sample preparation; titrate antibody concentration downward.

Frequently asked questions

What is the formulation of MAP3K5 Mouse mAb (Ab351000)?

MAP3K5 Mouse mAb (Ab351000) is formulated in PBS (Phosphate-Buffered Saline) only. This carrier-protein-free formulation is suitable for applications requiring direct conjugation or assays sensitive to additives such as BSA or sodium azide.

How should MAP3K5 Mouse mAb be stored to maintain its activity?

For long-term storage, keep MAP3K5 Mouse mAb at –20 °C. Avoid repeated freeze–thaw cycles by preparing single-use aliquots upon first thaw. For short-term use within one to two weeks, aliquots may be kept at 4 °C. Avoid prolonged exposure to room temperature or direct light.

What is the biological significance of MAP3K5 (ASK1) as a research target?

MAP3K5, also known as ASK1, is a serine/threonine kinase critical for stress-activated signaling pathways including the p38 MAPK and JNK cascades. It mediates cellular responses to oxidative stress, ER stress, and inflammatory cytokines, playing key roles in apoptosis, inflammation, and disease processes such as cancer, neurodegeneration, and cardiovascular conditions.

Which secondary antibody should be used with MAP3K5 Mouse mAb?

Since MAP3K5 Mouse mAb is produced in mouse, an anti-mouse secondary antibody conjugated to the appropriate reporter (e.g., HRP for Western blot or a fluorophore for immunofluorescence) should be used. Ensure the secondary antibody is validated for your specific application and detection system.

References

1. Ichijo H, et al. Induction of apoptosis by ASK1, a mammalian MAPKKK that activates SAPK/JNK and p38 signaling pathways. Science. 1997;275(5296):90-94.
2. Matsuzawa A, Ichijo H. Redox control of cell fate by MAP kinase: physiological roles of ASK1-MAP kinase pathway in stress signaling. Biochim Biophys Acta. 2008;1780(11):1325-1336.
3. Aladdin Scientific. Ab351000 MAP3K5 Mouse mAb Product Information Sheet. Aladdin Scientific; [current version].

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