Anti-MMP-14 Antibody, clone 1J5 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1760 - recombinant, expressed in HEK 293 cells
Synonym: Anti-MMP-14
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human colon (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1760, Anti-MMP-14, clone 1J5 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Clone 1J5 exhibited cytoplasmic/punctate cytoplasmic/membranous staining in glandular cells of human colon tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human colon (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1760, Anti-MMP-14, clone 1J5 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Clone 1J5 exhibited cytoplasmic/punctate cytoplasmic/membranous staining in glandular cells of human colon tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human colon (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1760, Anti-MMP-14, clone 1J5 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Clone 1J5 exhibited cytoplasmic/punctate cytoplasmic/membranous staining in glandular cells of human colon tissue sections.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HepG2 cells was performed using a 1:100 dilution of Cat. No. ZRB1760, Anti-MMP-14, clone 1J5 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the membrane, cytosol.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HepG2 cells was performed using a 1:100 dilution of Cat. No. ZRB1760, Anti-MMP-14, clone 1J5 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the membrane, cytosol.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HepG2 cells was performed using a 1:100 dilution of Cat. No. ZRB1760, Anti-MMP-14, clone 1J5 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the membrane, cytosol.
Western Blotting Enhanced Validation - Recombinant Antibody Technology Lysates from L929 cells (Lane 1) and A431 cells (Lane 2) were probed with Cat. No. ZRB1760, Anti-MMP-14, clone 1J5 ZooMAb® Rabbit Monoclonal (1:1,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates MMP-14 (~55 kDa).
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 100 mg/mL of non-specific control peptide (Run 1) or MMP14 peptide (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1760, Anti-MMP-14, clone 1J5 ZooMAb® Rabbit Monoclonal. The binding of this antibody to MMP14 peptide displayed a KD of 1.8 x 10-6, whereas binding to non-specific control peptide displayed a KD of 1.4 x 10-2.
Flow Cytometry Enhanced Validation - Recombinant Antibody Technology Staining of one million HepG2 cells was performed using 0.1 μg per million cells of Cat. No. ZRB1760, Anti-MMP-14, clone 1J5 ZooMAb® Rabbit Monoclonal (Yellow histogram) or the equivalent amount of a Rabbit IgG isotype control (Grey histogram), followed by a PE-conjugated Donkey Anti-Rabbit secondary antibody.
12 Principles of Green Chemistry: Principle 1—Waste Prevention. This product has a smaller environmental footprint through reduction of waste in the manufacturing process.
12 Principles of Green Chemistry: Principle 4—Designing Safer Chemicals
Chemical products should be designed to affect their desired function while minimizing their toxicity.
Chemical products should be designed to affect their desired function while minimizing their toxicity.
12 Principles of Green Chemistry: Principle 6—Design for Energy Efficiency. This product was re-engineered to utilize less energy in the manufacturing process.
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| biological source | rabbit |
| clone | 1J5, monoclonal |
| recombinant monoclonal | |
| conjugate | unconjugated |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| form | lyophilized |
| greener alternative category | Aligned |
| greener alternative product characteristics | Waste Prevention Designing Safer Chemicals Design for Energy Efficiency Learn more about the Principles of Green Chemistry . |
| isotype | IgG |
| mol wt | calculated mol wt ~65.89 kDa |
| observed mol wt ~55 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | mouse, human |
| storage temp. | 2-8°C |
| technique(s) | affinity binding assay: suitable |
| flow cytometry: suitable | |
| immunocytochemistry: suitable | |
| immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable | |
| western blot: suitable | |
| UniProt accession no. | P50281  |
| Application: | Quality Control Testing Evaluated by Western Blotting in L929 cell lysate. Western Blotting Analysis: A 1:1,000 dilution of this antibody detected MMP-14 in L929 cell lysate. Tested Applications Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected MMP-14 in A431 cell lysate. Affinity Binding Assay: A representative lot of this antibody bound MMP14 peptides with a KD of 1.8 x 10-6 in an affinity binding assay. Flow Cytometry Analysis: 0.1 μg from a representative lot detected MMP-14 in one million HepG2 cells. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected MMP-14 in human colon tissue sections. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected MMP-14 in HepG2 cells. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user |
| Application: | Anti-MMP-14, clone 1J5 ZooMAb®, Cat. No. ZRB1760, is a recombinant Rabbit monoclonal antibody that specifically targets MMP-14 and is tested in Affinity Binding Assay, Flow Cytometry, Immunocytochemistry, Immunohistochemistry, and Western Blotting. |
| Disclaimer: | Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals. |
| General description: | We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information. |
| General description: | ZooMAb antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb antibodies are reliably available and ready to ship when you need them. |
| Immunogen: | KLH-conjugated linear peptide corresponding to 16 amino acids from the extracellular domain of human Matrix metalloproteinase-14 (MMP14). |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant mouse monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 μL. |
| Reconstitution: | 300 μg/mL after reconstitution at 25 μL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type. |
| Specificity: | Clone 1J5 is a ZooMAb® rabbit recombinant monoclonal antibody that specifically detects Matrix metalloproteinase-14 (MMP-14). It targets an epitope within 16 amino acids from the extracellular domain. |
| Storage and Stability: | Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 μL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws. |
| Target description: | Matrix metalloproteinase-14 (UniProt: P50281; also known as EC:3.4.24.80, MMP-14, MMP-X1, Membrane-type matrix metalloproteinase 1, MT-MMP 1, MTMMP1, Membrane-type-1 matrix metalloproteinase, MT1-MMP, MT1MMP) is encoded by the MMP14 gene (Gene ID: 4323) in human. MMPs are a family zinc-dependent endoproteinases that degrade collagen and other structural proteins and are responsible for much of the turnover of matrix components. Each MMP contains a protease domain with a conserved HExGHxxGxxHS/T sequence, where three histidine residues form a complex with a catalytic Zn atom. A regulatory domain with a conserved PRCGxPD motif helps to maintain MMPs in their latent form by binding of the cysteine residue to the active site Zn. MMP-14 is a single-pass type I membrane protein that is expressed in stromal cells of colon, breast, head, and neck. It is a membrane type matrix metalloproteinase (MT-MMP) that is synthesized with a signal peptide (aa 1-20) and a propeptide (aa 21-111) that are subsequently cleaved off to generate the active mature form. The propeptide maintains it in a latent state. The mature enzyme contains an extracellular domain (aa 112-541), a transmembrane domain (aa 542-562), and a cytoplasmic domain (aa 563-582). MMP14 contains four hemopexin domains within its extracellular domain and a cysteine switch (aa 91-98). The conserved cysteine present in the cysteine-switch motif binds the catalytic zinc ion, thus inhibiting the enzyme. The dissociation of the cysteine from the zinc ion upon the release of propeptide activates the enzyme. This ZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. |
| WGK Germany | WGK 1 |
| Storage Temp. | 2-8°C |