|Product Name||Anti-MMP16 Antibody|
|Storage & Handling||At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time.Avoid repeated freezing and thawing.|
|Description||Rabbit IgG polyclonal antibody for Matrix metalloproteinase-16(MMP16) detection. Tested with WB, IHC-P, IHC-F in Human;Mouse;Rat.|
|Cite This Product||Anti-MMP16 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1123)|
|Immunogen||A synthetic peptide corresponding to a sequence at the C-terminus of human MMP16(582-598aa, YTVFQFKRKGTPRHILY), identical to the related rat and mouse sequences.|
|Reactivity||Human, Mouse, Rat|
Assay Dilutions Overview
Immunohistochemistry(Frozen Section), 0.5-1μg/ml, Human, Rat, Mouse
Immunohistochemistry(Paraffin-embedded Section), 0.5-1μg/ml, Human, Rat, Mouse, By Heat
Western blot, 0.1-0.5μg/ml, Human, Rat, Mouse
Boster's Secondary Antibodies And IHC, WB Kits
The following reagents are used to generate the images below.Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(P) and IHC(F).
Images And Assay Conditions
Figure 1. Western blot analysis of MMP16 using anti-MMP16 antibody (PA1123).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human COLO-320 whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-MMP16 antigen affinity purified polyclonal antibody (Catalog # PA1123) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for MMP16 at approximately 53KD. The expected band size for MMP16 is at 69KD or 53KD.
Figure 2. IHC analysis of MMP16 using anti- MMP16 antibody (PA1123).
MMP16 was detected in paraffin-embedded section of human rectal cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti- MMP16 Antibody (PA1123) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Protein Target Info (Source: Uniprot.org)
|Protein Name||Matrix metalloproteinase-16|
|Tissue Specificity||Expressed in heart, brain, placenta, ovary and small intestine. Isoform Short is found in the ovary.|
|Alternative Names||Matrix metalloproteinase-16;MMP-16;3.4.24.-;MMP-X2;Membrane-type matrix metalloproteinase 3;MT-MMP 3;MTMMP3;Membrane-type-3 matrix metalloproteinase;MT3-MMP;MT3MMP;MMP16;MMPX2;|
|Subcellular Localization||Isoform Long: Cell membrane ; Single-pass type I membrane protein ; Extracellular side . Localized at the cell surface of melanoma cells.|
|Molecular Weight||69521 MW|
*if product is indicated to react with multiple species, protein info is based on the human gene.
|Protein Function||Endopeptidase that degrades various components of the extracellular matrix, such as collagen type III and fibronectin. Activates progelatinase A. Involved in the matrix remodeling of blood vessels. Isoform short cleaves fibronectin and also collagen type III, but at lower rate. It has no effect on type I, II, IV and V collagen. However, upon interaction with CSPG4, it may be involved in degradation and invasion of type I collagen by melanoma cells. .|
|Research Areas||Invasion/Microenvironment, Metalloprotease, Proteolysis / Ubiquitin, Proteolytic Enzymes
*You can search these to find other products in these research areas.
|Background||The matrix metalloproteinase 16(MMP16) protein consists of 604 amino acids and has a characteristic MMP domain structure, which gene is mapped on human chromosome 8q21. Additionally, MMP16 has a C-terminal extension containing a potential transmembrane domain, similar to MMP14 , MMP15 , and MMP17. Furthermore, it is membrane-bound and is a member of the membrane-type MMPs that are a subclass in the MMP family since the other members lack a C-terminal transmembrane domain and are secreted as soluble forms. MMP16 is expressed as a 12-kb transcript in brain, placenta, heart, and some carcinoma cell lines, but is not detectably expressed in lung, kidney, liver, spleen, and muscle.|
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1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected
2. Post-translational cleavage: this can cause smaller bands and or multiple bands
3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody.
4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.
5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher.,