Anti-MMP-27 Rabbit Polyclonal Antibody

Frequently Asked Questions

What molecular weight band should I expect for MMP-27 on Western blot?

MMP-27 has a predicted molecular weight of approximately 57 kDa based on its 513-amino-acid sequence. On Western blot, you should expect a band near this size, though post-translational modifications including glycosylation may shift the apparent molecular weight slightly higher. Because MMP-27 is retained in the endoplasmic reticulum rather than secreted, ensure you are using whole-cell lysates rather than conditioned media. If you observe multiple bands, consider that partial proteolytic processing or splice variants may be present depending on your cell type and lysis conditions.

What starting dilution should I use for MMP-27C antibody in Western blot?

We recommend starting at 1:1000 dilution for Western blot applications with this rabbit polyclonal antibody. This dilution has been validated in human samples and provides a good balance between signal and background in most lysate types. Depending on MMP-27 expression levels in your sample, you may need to optimize between 1:500 and 1:2000. Use lysates from cells or tissues known to express MMP-27 as positive controls, and block membranes with five percent non-fat milk or BSA in TBST for optimal results.

Is MMP-27 localized intracellularly or secreted into culture media?

Unlike most matrix metalloproteinases, MMP-27 is retained in the endoplasmic reticulum and is not secreted into the extracellular space or culture supernatant. This ER localization distinguishes MMP-27 from classical secreted MMPs and suggests it has specialized intracellular proteolytic functions. When designing experiments, harvest whole-cell lysates rather than relying on conditioned media. For immunofluorescence or immunohistochemistry applications, expect perinuclear staining consistent with ER localization. This retention pattern is important when interpreting MMP-27 biology in matrix remodeling contexts.

Does the MMP-27C antibody cross-react with mouse or rat samples?

This antibody has been validated specifically for human MMP-27 detection. Cross-reactivity with mouse or rat MMP-27 has not been experimentally confirmed. Human MMP-27 shares moderate sequence homology with rodent orthologs, so cross-reactivity is possible but should be considered predicted rather than validated. If you are working with mouse or rat samples, we recommend running a positive control using human lysate alongside your experimental samples to assess whether specific signal is detectable. Alternatively, request species-specific validation data or consider antibodies explicitly validated for rodent MMP-27.

What positive control cell line or tissue should I use for MMP-27 expression?

MMP-27 expression is relatively restricted compared to other MMPs, so selecting an appropriate positive control is important. Published literature indicates MMP-27 mRNA expression in tissues including liver, kidney, and certain carcinoma cell lines, though protein-level validation varies. If your target cells or tissues have unknown MMP-27 status, consider running a small panel of human cancer cell lines or normal tissue lysates alongside your samples. Negative controls using MMP-27 knockout cells or siRNA knockdown lysates can help confirm antibody specificity and distinguish true signal from non-specific background.

Can I use this antibody for immunohistochemistry on paraffin-embedded sections?

Triple Point Biologics antibodies are typically validated for Western blot and immunohistochemistry applications, and this rabbit polyclonal format is suitable for IHC. For paraffin-embedded sections, antigen retrieval is usually required. Start with citrate buffer pH 6.0 heat-induced epitope retrieval and a 1:100 to 1:200 antibody dilution. Because MMP-27 is ER-localized, expect perinuclear cytoplasmic staining rather than extracellular or nuclear signal. Optimize retrieval conditions and dilutions with positive control tissue known to express MMP-27, and include isotype or pre-immune serum controls to assess non-specific binding.

How should I store the MMP-27C antibody to maintain stability?

Store the antibody at minus twenty degrees Celsius for short-term use or minus eighty degrees Celsius for long-term storage. Avoid repeated freeze-thaw cycles, which can reduce antibody activity and increase aggregation. For frequent use, aliquot the antibody upon receipt into single-use volumes to minimize freeze-thaw exposure. The antibody is supplied in a stabilized buffer; do not dilute the stock solution for storage. Once diluted in blocking buffer for Western blot or IHC, use the working solution the same day or store at four degrees Celsius with sodium azide preservative for up to one week.

What sample preparation is critical for detecting intracellular MMP-27 by Western blot?

Because MMP-27 is ER-resident, thorough cell lysis is essential to solubilize membrane-associated protein. Use RIPA buffer or similar detergent-containing lysis buffer with protease inhibitor cocktail to prevent proteolytic degradation during extraction. Avoid using conditioned media, as MMP-27 is not secreted. Sonication or freeze-thaw cycles can improve yield from tough tissues. Load twenty to fifty micrograms total protein per lane as a starting point, and include reducing agent in your sample buffer since MMP-27 may form disulfide-linked complexes. Include a positive control lysate to confirm lysis efficiency and antibody performance.