rev: September 16, 2002

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  ANTIBODIES  

(anti-Human and others as indicated)

RDI Divison of Fitzgerald Industries Intl  offers a wide line of  antibodies. Since no one antibody works best for all applications (neutralization, blotting, ELISA, etc), we offer many different types of antibodies to help solve this problem. Please inquire for other applications or types of antibodies not listed below.


MMP-14/MT1-MMP Antibodies

see also anti-MMP-1, MMP-2, MMP-3, MMP-7 , MMP-8, MMP-9 and MMP-13, MT2-MMP (MMP15) , MT3-MMP


Matrix metalloproteinases (MMPs) are a family of enzymes that are responsible for the degradation of extracellular matrix components. Of the eleven proteins reported to date, ten are normally found as soluble molecules. The eleventh, and newest of these proteins, has been named MT-MMP for membrane bound MMP. MT-MMP contains a C-terminal transmembrane domain which allows it to localize to the cell surface. It is also unique from the other members of the MMP family in that it contains an 8 amino acid insert in the catalytic domain. The protein is encoded by a 4.5 kb mRNA species giving rise to a protein with a molecular weight of 60-66 kDa by SDS-PAGE. MT-MMP is responsible for cleaving gelatinase A (MMP-2, 72 kDa Type IV collagenase) to the active form.Evidence exist to suggest that MT-MMP itself requires an activation step which is most likely to result of activity of the membrane plasmin cascade. MT-MMP functions by binding TIMP-2 and then the COOH terminal end of MMP-2 resulting in a 105 kDa trimer which effects the cleavage of pro-MMP-2 to the biologically active form. The order of the binding of pro-MMP-2 and TIMP-2 to MT-MMP is critical as TIMP-2 will also inhibit the activity of MMP- when present in a soluble form. Since its discovery,a second form of MT-MMP has been identified resulting in a change in the name of the original protein to MT-MMP-1.

clone 113-5B7   cat#RDI-MT1MMPabm-B7    $438.00/100ug

(suitable for use on human, mouse, rat and rabbit), mIgG3 isotype

-reactive in histochemistry (paraffin sections, frozen sections, use lung or breast carcinoma tissue , western blot (MD-MB-231 cells pretreated with 1-5ug/ml ConA for 24-48 hours,  IP, immunofluorescence). see aa 319-333 in the hemopexin-like domain of human MT1-MMP (slightly cross reacts with human MMP-3).


clone 114-6G6  cat#RDI-MT1MMPabm-G6    $438.00/100ug

suitable for use on human), mIgG1

-reactive in histochemistry paraffin sections (not frozen sections), western blot ,not for IP use.

-sees aa 160-173 in the catalytic domain of human MT1-MMP.


DATA SHEET:Mouse anti-human MT1-MMP(MMP-14) monoclonal     clone 114-1F2        (WB/IP)

Catalog#: RDI-MT1MMPabm     Price: $438.00/vial  


Package Size: 100ug (lyophilized)   with 100ug BSA.


Dilution: Tighten cap, tap vial to settle contents. Reconstitute with 1ml sterile PBS pH 7.4 or sterile 20mM Tris-saline (20mM Tris containing 0.15M NaCl) pH 7.4. allow to set 2-3 hours at 4 DEG C, mix well. Addition of 0.1% sodium azide recommended or other preservative if suitable for your system (do not add azide if to be used in neutralization type reactions)

Ig Isotype: mouse IgG1

clone: 114-1F2

Antigen: peptide corresponding to amino acids 160-173 of human MT-MMP-1

Reactivity: Binds to human MT-MMP-1

Suitable for: western blotting (approx 10ug/ml)using enhanced cheiluminescence detection, positive control HT-1080 cells. or   ConA treated MD-MB231 cells (treat with 1-   5ug/ml ConA for 24-48hours)

                    -immunoprecipitation

Ref: -Sato, H et al (1994) Nature 370:61-65

       -Yu, M et al (1995) Cancer Res 55:3272-3277

Storage: Store 4 DEG C or lower. Avoid frequent freeze thaw cycles. Antibody is stable for at least 6 months when stored frozen undiluted and up to 1 month at 4 DEG C when neat or diluted as per above recommendation. Recommend adding 0.1% azide if suitable for your assay system. Dilution buffers should contain 0.1-1% BSA.


Precautions: For In vitro research Use Only. Not for use in or on humans or animals or for diagnostics. It is the responsibility of the user to comply with all local/state and Federal rules in the use of this product. We are not responsible for any patent infringements that might result with the use of or derivation of this product.

-copyright by owner


Rabbit Anti-MMP-14 (MT1-MMP)  (Human/Mouse) Hemopexin domain

cat# RDI-MMP14HEabr   $469.00

Presentation: 50ug Protein A purified rabbit IgG in 0.0185ml (2.7mg/ml) 10mM PBS, pH 7.4, with 15mM sodium azide.

IMMUNOGEN: Generated against the complete human hemopexin-like domain (recombinant).

SPECIFICITY: recognizes a 60-66 kDa protein identified as MTI-MMP or MMP14.

LOCALIZATION: Cell Membrane

Use: Immunohistochemistry: 1:100-1:500 When used for immunohistochemical staining of mouse brain cryosections, the antibody works best for tissues fixed with ice-cold acetone (10-15 minute fix) or Carnoy’s solution (10-15 minute fix). The antibody does not work for formaldehyde fixed cryosections. After fixation, slides may be washed with PBS/0.1% BSA for 3 x 5 minutes, followed by incubation with the anti-MT-MMP antibody at a 1:100 - 1:500 dilution for 90 minutes. Following 3 x 5 minute washes with PBS, and 1 x 5 minute wash with PBS/0.1% BSA, incubate with appropriate secondary antibody and detect.

Immunoblotting: 1:500 - 1:2,000. Suggested incubation conditions with nitrocellulose membranes include incubation of blot strips in 3 mL PBS or 50 mM Tris-HCl buffer, pH 7.5 - 8.0 containing either dry milk powder or 1% BSA and 1:500 - 1:2,000 dilution of antibody. Incubate for 2-4 hours at 4C. Overnight incubations may result in higher background when membrane fractions or whole cell lysates are analyzed.

Immunocytochemistry of transfected cells: 1:100-1:600

FACS analysis: 1:1000

Immunoprecipitation: from cell lysates containing 1-2% Triton X-100 or NP40. SDS concentrations of 0.1 - 0.2% do not interfere with the precipitation. Immune complexes may be isolated on Protein A-Separose or agarose.

Optimal working dilutions must be determined by the end user.

POSITIVE  CONTROL: HFL-1 Cells, Placenta

SPECIES REACTIVITIES: Human, Mouse. Others not tested.

STORAGE: Store vial at 4ºC. When stored at 2-8ºC, this antibody is stable for 12 months.

FOR RESEARCH USE ONLY

Note: Upon receipt, tighten cap and centrifuge for 1-2 minutes at1000RPM to concentrate material in vial.


Rabbit anti-MT-MMP-1 (MMP-14)

(Membrane-type Matrix Metalloproteinase-1)(Hinge region)

CAT#: RDI-MMP14Habr    $469.00

Quantity: 100ug peptide-affinity purified immunoglobulin   in PBS with sodium azide or thimerosal.

Immunogen: Synthetic peptide based on the human sequence.

Specificity: Binds to MT-MMP-1, No cross reactivity with other MMP family members (MMP-1, MMP-2, MMP-3, MMP-9).

Applications: Works well for Western Blotting applications, identifying reduced protein bands at 65 and 63 kDa (Proenzyme and active MT-MMP-1), and further processed species. The antibody recognizes native MT-MMP-1, for application in immunoprecipitation, immunohistochemistry, ELISA and cell sorting. -MT-MMP-1, expression is elevated in several tumor cell lines, and constitutive in some normal cell lines. Concanavalin A or TPA will stimulate MT- MMP-1 production in some cell types, and shed forms of the enzyme have been reported.


Dilution: A recommended starting dilution for Western Blots is 1:2,000 when using colorimetric substrates such as BCIP/NBT, and 1:5,000 for chemiluminescent substrates. Higher concentration of antibody may be required for non-human samples. Optimal working dilutions must be determined by the end user.


Storage: The undiluted antibody solution is stable for approximately 6 months when stored between 2 and 8'C. For longer storage, mix antibody solution with glycerol 1:1 to prevent freezing, and store at -20'C. Diluted antibody can be used several times if not contaminated; store between 2 and 8'C with 0.01% sodium azide as preservative.

FOR RESEARCH USE ONLY


DATA SHEET: Mouse anti-human MT1-MMP(MMP-14) monoclonal   (WB/IP) HUMAN/Mouse/Rat/Rabbit

Catalog#: RDI-MT1MMPabm-B7  Price: $438.00/vial

Package Size: 100ug in 1ml 0.05M sodium phosphate with 50% glycerol.

Ig Isotype: mouse IgG3k

clone: 113-5B7

Antigen: peptide corresponding to amino acids 319-333 of human MT-MMP-1 (hemopexin-like domain).

Reactivity: Binds to human MT-MMP-1, also rat, mouse, and rabbit (slight cross with human MMP-3).

Suitable for: western blotting (approx 10ug/ml)using enhanced cheiluminescence detection, positive control HT-1080 cells, or ConA treated MD-MB231 cells (treat with 1- 5ug/ml ConA for 24-48hours), sees band approx 58kda

-immunoprecipitation

-frozen, paraffin sections and immunofluorescence (see Nature 370:61-65 (1994) lung carcinoma as pos control.

Ref: -Sato, H et al (1994) Nature 370:61-65

       -Yu, M et al (1995) Cancer Res 55:3272-3277

Storage: Store -20 DEG C . Avoid frequent warming cycles. Antibody is stable for at least 12 months when at -20 DEG C.Recommend adding 0.1% azide if suitable for your assay system. Dilution buffers should contain 0.1-1% BSA.

It is the responsibility of the user to comply with all local/state and Federal rules in the use of this product. We are not responsible for any patent infringements that might result with the use of or derivation of this product.

see also anti-MMP-1, MMP-2, MMP-3, MMP-7 ,MMP-8, MMP-9 ,MMP-13, MMP-14, MMP15 & MMP16


RDI Divison of Fitzgerald Industries Intl

34 Junction Square Drive

Concord MA 01742-3049

USA

phone (800) 370-2222

     or  (978) 371-6446 or (800) 370-2222

fax     (978) 371-2266

EMAIL:antibodies@fitzgerald-fii.com

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