Anti-His Tag Mouse Monoclonal Antibody (5C3)

His tag (commonly 6×His or 8×His) is widely used for recombinant protein purification and detection. Anti-His antibodies are useful for confirming expression and tracking the tagged protein across purification and assays.

Why buyers use anti-His

• WB verification: Confirms expression and expected size of His-tagged proteins.
• Purification tracking: Helps identify which fractions contain the His-fusion during purification.
• General detection: Useful when a native antibody is unavailable for the target protein.

When to consider another approach

• Tag position/accessibility: N- vs C-terminal placement and folding can affect binding.
• Native vs denatured conditions: verify the antibody is validated for your assay conditions.

Clone 5C3 is the unique monoclonal antibody clone designation for this product. If you previously used a His Tag antibody labeled “5C3,” this clone number is the key identifier that helps you select the same antibody lineage for better experiment-to-experiment consistency.

What “Clone 5C3” tells you (and what it doesn’t)

• It tells you: this antibody comes from a single monoclonal clone (5C3), meaning the antibody population is uniform and targets one dominant epitope/epitope region.
• It does not guarantee: identical performance across all suppliers, because formulation, purification, conjugation, and validation standards can differ even when the clone number matches.

When choosing Clone 5C3 is especially helpful

• Replacing an antibody: If your previous antibody was “His Tag (Clone 5C3),” matching the clone is the best first step to keep band patterns and background similar.
• Protocol/paper matching: Many published protocols specify the clone. Using the same clone reduces variability when reproducing literature results.
• Standardization across projects: Clone-based standardization is useful when multiple people or sites run the same workflows.

Buyer checks before committing

• Application validation: Confirm Clone 5C3 is validated for your intended assay(s) listed on the product page (e.g., WB vs IF/IHC vs IP).
• Specificity expectations: Review validation images (if available) and confirm the expected signal pattern for your sample type.
• Plan controls: If you are switching from a different clone, run side-by-side comparisons and include positive/negative controls to confirm specificity and background.

For WB, the apparent band size depends on your fusion protein (tag + protein of interest) and sample conditions.

Tip: If you need faster WB or IP workflows, look for HRP- or bead-conjugated formats of the same clone when available.

Monoclonal antibodies are produced from a single B-cell clone and recognize one dominant epitope/epitope region on the target. They are often chosen for their consistent performance and lot-to-lot reproducibility.

Why buyers choose monoclonal antibodies

• Reproducibility: More consistent results across experiments and over time.
• Specificity: Typically cleaner signals when the epitope is well exposed.
• Standardization: Easier to match literature protocols when a clone is specified.

When to consider another option

• if the target epitope is masked by fixation, folding, or complex formation
• if you need broader recognition across isoforms or modified forms of the protein

In these cases, a polyclonal antibody can sometimes provide stronger overall signal because it recognizes multiple epitopes.

Application(s) indicate which experimental methods this antibody has been tested/validated for. Choosing an antibody validated for your specific method is one of the best ways to reduce troubleshooting.

What the listed application(s) mean

• Western blot (WB): Detects proteins in denatured samples separated by gel electrophoresis; ideal for confirming size and expression changes.
• Immunofluorescence (IF): Fluorescent staining in cells/tissues; fixation/permeabilization choices can impact epitope accessibility.
• Immunoprecipitation (IP): Enriches target proteins (and sometimes complexes) from lysates; antibody format and bead chemistry affect yield and background.

Buyer guidance

• Match your workflow: If your main use is WB, IF, IHC-P, or IP, prioritize antibodies validated for that exact application.
• Sample prep matters: Fixation, permeabilization, antigen retrieval, and lysis buffer can change epitope accessibility and background.
• Use controls: Include positive/negative controls and, when possible, compare with a known antibody or sample.

Host refers to the species in which the primary antibody was raised (here: Mouse). Host selection mainly affects which secondary antibody you should use and can influence background in certain sample types.

Why buyers care about the host

• Secondary antibody matching: Choose an anti-mouse secondary (e.g., anti-mouse IgG) with the right conjugate for your detection method.
• Background control: If your sample contains endogenous immunoglobulins from the same species as the host, background can increase (common in some tissue IHC workflows).
• Multiplex design: Using primaries from different host species helps avoid cross-reactivity in multi-target staining.

When to take extra care

• Mouse-on-mouse IHC/IF (mouse primary on mouse tissue) can increase background; consider mouse-on-mouse kits, highly cross-adsorbed secondaries, or alternative host species.
• For complex staining panels, use cross-adsorbed secondaries and include no-primary controls to check nonspecific binding.

Reactivity describes which species the antibody is expected to recognize based on validation and/or target sequence conservation. Reported reactivity for this product includes: Mammals, Bacteria.

Why buyers should check reactivity

• Species compatibility: Make sure your sample species is included in the reported reactivity list before ordering.
• Sequence conservation: Even when species are closely related, small sequence differences can affect binding (especially for monoclonals).
• Sample context: Tissue type, expression level, and sample prep can influence whether reactivity translates into a clean signal.

Buyer guidance

• If your species is not listed, consider requesting additional validation or testing a small pilot first.
• Use a positive control sample from a listed species whenever possible to confirm performance.
• For tag antibodies (e.g., GFP/HA/FLAG), reactivity often reflects where the tag is used rather than a native species protein—fusion design and expression level still matter.

Unconjugated primary antibodies are supplied without a detection label. This is the most flexible format and can be paired with different secondary antibodies (HRP, fluorophores, biotin, etc.) depending on your assay.

Why buyers choose unconjugated antibodies

• Maximum flexibility: Use the same primary with different detection systems across WB, IF, or IHC.
• Signal tuning: Choose a secondary antibody that matches your sensitivity and multiplex needs.
• Cost efficiency: One primary can support multiple workflows by switching secondaries.

When to consider a conjugated format

• If you want fewer steps and faster WB, consider an HRP-conjugated primary.
• If your main workflow is IP/Co-IP, bead-conjugated formats (agarose or magnetic beads) can simplify pull-down and reduce background.