rev:  March 14, 2005

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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.


TIMP (Tissue Inhibitors of Metalloproteinase) Antibodies

-see below for polyclonals and moncolonals


Polyclonals suitable for immunoblotting: ($438.00/vial 3+  any combination)

cat#RDI-TIMP1abr  rabbit anti-human TIMP-1  100ug  $469.00

cat#RDI-TIMP2abr  rabbit anti-human TIMP-2 (hu, rat, mouse, bovine)  $469.00/100ug

cat#RDI-TIMP3abr  rabbt anti-human TIMP-3      $469.00/100ug

cat#RDI-TIMP4abr  rabbit anti-human TIMP-4     $469.00/100ug

-see also goat polyclonals below


DATA SHEET:   Mouse anti-human TIMP-1 monoclonal     (WB/Histochemistry)

Catalog#: RDI-TIMP1-C1   Price: $438.00

Package Size: 100ug (lyophilized)

Ig Isotype: mouse IgG1

clone: 7-6C1

Reactivity: Binds to TIMP-1 from human, bovine and rat (not mouse)

Suitable for: western blotting-approx 1ug/ml with enhanced chemiluminescence detection, sees band at approx 30kda.

                  -histochemistry (frozen sections, not paraffin)

Note:TIMP-1 is found in very low concentrations in most cells and tissues. Use of amplified detection systems is recommended,

Positive Control: Phorbol ester treated HT 1080 conditioned medium OR rec TIMP-1


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.


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.

recombinant human TIMP-1 cat#RDI-TIMP1-AG   $344.00/3ug

>98% pure by SDS=PAGE, suitable for use in WB and competition assays. MW=28kDa


mouse anti-TIMP-1    cat#RDI-TIMP1-6D11   (WB/Paraffin)   $438.00/100ug lyophilized

Package Size: 100 µg purified antibody lyophilized from a volatile   buffer with 100 µg of BSA. We recommend resuspending the   lyophilized antibody with sterile phosphate buffered saline   (PBS), pH 7.4, or sterile 20 mM Tris-saline (20 mM Tris   containing 0.15 M NaCl), pH 7.4, to yield a final   concentration of 100 µg/mL; product will be more stable if   0.1% sodium azide is included (do not add azide if antibody   is to be used with viable cells). Lyophilized antibodies   should be resuspended at 4°C with occasional gentle   mixing for at least two hours.

Ig Isotype: mouse IgG1k

clone: 147-6D11

Immunogen: recombinant TIMP-1 protein and fusing the splenocytes with NS1 mouse myeloma cells.

Reactivity: Binds to TIMP-1 from human only

Suitable for: western blotting-approx 1ug/ml with enhanced chemiluminescence detection,
                 -histochemistry -paraffin sections, titter from 10ug/ml- 1ug/ml with ABC/Dab detection.

Epitope: Not determined

STORAGE Store at 4°C until reconstituted, then store in aliquots at -20°C or at 4°C with 0.1% azide. Freezing of aliquots is best for storage of reconstituted product for longer than a month; repetitive freezing and thawing should be avoided. If stored under proper conditions, product guaranteed until expiration date stated.

Matrix metalloproteinases (MMPs) are a family of enzymes that are responsible for the degradation of extracellular matrix components such as collagen, laminin and proteoglycans (1-4). In addition to sequence homology, all MMPs share the following characteristics: the catalytic mechanism is dependent upon zinc ion at the active center, they cleave one or more extracellular matrix components, they are secreted as zymogens which are activated by removal of an approximately 10 kDa segment from the N-terminus and their activity is regulated by endogenous inhibitors. These enzymes are involved in normal physiological processes such as embryogenesis and tissue remodeling and may play an important role in arthritis, periodontitis, and metastasis. The activation and activity of MMPs are regulated by a family of endogenous inhibitors, tissue inhibitiors of metalloproteinase (TIMP). TIMP-1 is a 28 kDa glycoprotein that is expressed by a variety of cell types. It forms a non-covalent, stoichiometric complex with both latent and active MMPs. TIMP-1 preferentially binds and inhibits MMP-9 and MMP-1 protein (5). TIMPs are capable of altering the metastatic potential of cancer cells and have been shown to inhibit invasion and metastasis in animal models as well as to be subject to differential expresssion in other disease states (6-8).

GENERAL REFERENCES

1. Liotta, L. A. and Stetler-Stevenson, W. G. in Seminars in Cancer Biology, ed. M. M. Gottesman. Vol. 1(2); 99-106, 1990.

2. Cottam, D. W. and Rees, R. C. Intl J. Oncol. 2; 861-872, 1993.

3. Woessner, J. F. FASEB J. 5; 2145-2154, 1991.

4. Stetler-Stevenson, W. G., et al. FASEB J. 7; 1434-1441, 1993.

5. Vallon, R., et al. Eur. J. Biochem. 244: 81-88, 1997.

6. Rivera, S., et al. J. Neurosci. 17: 4223-4235, 1997.

7. Herbst, H., et al. Am. J. Pathol. 150: 1647- 1659, 1997.

8. Jung, K., et al. Int. J. Cancer 74: 220-223,1997.

For In Vitro Research Use Only


DATA SHEET:   Mouse anti-human TIMP-2 monoclonal    (WB/Immunoprecipitation)

Catalog#: RDI-TIMP2-T2    Price: $438.00/vial  

Package Size: 100ug (lyophilized)

Ig Isotype: mouse IgG1

clone: T2-101

Reactivity: Binds to TIMP-2 from human (not mouse or rat)

Suitable for: western blotting-approx 1ug/ml with enhanced chemiluminescence detection, sees band at approx 24kda.

                 -histochemistry (frozen sections, not paraffin sections)

Note:TIMP-2 is found in very low concentrations in most cells and tissues. Use of amplified detection systems is recommended,

Positive Control: recombinant human TIMP-2

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.


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.


recombinant human TIMP-2 cat#RDI-TIMP2-AG $281.00/3ug

>98% pure by SDS=PAGE, suitable for use in WB (approx 300ng/lane) and competition assays. MW=24kDa


mouse anti-human TIMP-2  (Paraffin section/Western blot)

cat#RDI-TIMP2-4H11  clone 67-4H11,  $438.00/100ug (lyophilized)

Package Size: 100ug (lyophilized) with 100ug BSA. Recommend   reconstituting the antibody with sterile PBS,   pH 7.4 or sterile 20mM Tris saline (20mM Tris   containing 0.15M NaCl), pH 7.4 to yield a   100ug/ml antibody concentration. Allow to   reconstitute for at least 2 hours with gentle mixing prior to use.

Ig Isotype: mouse IgG1k

clone: 67-4H11

Immunogen: synthetic peptide corresponding to amino acids 178-193 of human TIMP-2 (numbered from the propeptide)

- Y R G A A P P K Q E F L D I E D

Reactivity: -Binds to TIMP-2 from human, mouse, rat rabbit, bovine, and guinea pig

-recognizes a 24kDa band by western blot, approx 5ug/ml with enhanced chemiluminescence detection.
-histochemistry (frozen sections, approx 1ug/ml) & paraffin sections approx 5ug/ml)

ref: Tomita T. and Iwata, K (1996) Dis Colon Rectum 39:1255-1264.

Positive Control: recombinant human TIMP-2

general ref: J. Immunol Meth 187:33-39 (1995)

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. For prolonged storage add 0.1% azide. Dilutions in a buffer containing 0.1- 1% BSA are recommended.


DATA SHEET: Mouse anti-human TIMP-3 monoclonal   (WB) Paraffin Sections

Catalog#: RDI-TIMP3-C1    Price: $438.00/vial

Package Size: 100ug antibody lyophilized from a volatile buffer with 100 µg of BSA. We recommend resuspending the lyophilized antibody with 1ml sterile phosphate buffered saline (PBS), pH 7.4, or sterile 20 mM Tris-saline (20 mM Tris containing 0.15 M NaCl), pH 7.4, to yield a final concentration of 100ug/ml. Stable if 0.1% sodium azide is included (do not add azide if antibody is to be used with viable cells). Lyophilized antibodies should be resuspended at 4°C with occasional gentle mixing

Ig Isotype: mouse IgG1

clone: Clone 136-13H4 is a mouse monoclonal antibody generated by immunizing mice with a peptide corresponding to amino acids 170-188 of human TIMP- 3 and fusing the splenocytes with NS1 mouse myeloma cells (see ref. 7).

REACTIVITY: Positive: Human, baboon, rabbit

                          Negative: frozen sections of rat, cow, pig and chicken (7)

TIMP-3 (clone 126-13H4) will recognize both glycosylated (27 kDa) and unglycosylated (24 kDa) TIMP-3 (7). This antibody does not cross-react with either   human TIMP-1 or TIMP-2. Suggested starting   concentrations are provided. Antibodies should be titrated for optimal results in individual system. In general, use higher concentrations to enhance sensitivity and lower concentrations to enhance specificity.


USE: Western Blot 5 µg/mL

Frozen Sections 2-10 µg/m (pos ocntrol unfixed eye)

Paraffin Sections 10 µg/mL   Heat release  (see ref. 7)

Positive Control: 293 or CACo-2 cells or eye

ref: Fariss, R.N. et al. (1997) Am. J. Pathol. 150: 323-328. (Frozen sections, paraffin sections and western blotting)

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.

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.

BACKGROUND

Matrix metalloproteinases (MMPs) are a family of enzymes that are responsible for the degradation of extracellular matrix components such as collagen, laminin and proteoglycans (1-4). In addition to sequence homology, all MMPs share the following characteristics: the catalytic mechanism is dependent upon a zinc ion at the active center, they cleave one or more extracellular matrix components, they are secreted as zymogens which are activated by removal of an approximately 10 kDa segment from the N-terminus and their activity is regulated by endogenous inhibitors. These enzymes are involved in normal physiological processes such as embryogenesis and tissue remodeling and may play an important role in arthritis, periodontitis, and metastasis. The activation and activity of MMPs are regulated by a family of endogenous inhibitors, tissue inhibitiors of metalloproteinase (TIMP). TIMP-3 is a 27 kDa glycoprotein (24 kDa unglycosylated) extracellular-matrix associated protein that is expressed by a variety of cell types (5). It forms a non-covalent, stoichiometric complex with both latent and active MMPs. TIMP-3 inhibits activation of progelatinase A by MT2-MMP (6). TIMPs are capable of altering the metastatic potential of cancer cells and have been shown to inhibit invasion and metastasis in animal models as well as to be subject to differential expresssion in other disease states. Expression of TIMP-3 is quite marked in the eye and it has recently been shown that Sorsby's fundus dystrophy, a dominant inherited form of blindness is caused by mutation in the gene encoding TIMP-3 (7-10).

APPLICATION REFERENCES

Fariss, R.N. et al. (1997) Am. J. Pathol. 150: 323-328. (Frozen sections, paraffin sections and western blotting)

GENERAL REFERENCES

1. Liotta, L. A. and Stetler-Stevenson, W. G. in Seminars in Cancer Biology, ed. M. M. Gottesman. Vol. 1(2); 99-106, 1990.

2. Cottam, D. W. and Rees, R. C. Intl J. Oncol. 2; 861-872, 1993.

3. Woessner, J. F. FASEB J. 5; 2145-2154, 1991.

4. Stetler-Stevenson, W. G., et al. FASEB J. 7; 1434-1441, 1993.

5. Kishnani, N.S., et al. (1995) Matrix Biol 14 (6): 479-488.

6. Butler, G.S., et al. (1997) Eur. J. Biochem 244: 653-657.

7. Fariss, R.N., et al. (1997) Am. J. Pathol. 150: 323-328.

8. Ruiz, A., et al. (1996) Biochem. Biophys. Res. Commun. 226: 467-474.

9. Anand-Apte, B., et al. (1997) Invest. Ophthalmol. Vis. Sci. 38: 817-823.

10. Vranka, J.A., et al. (1997) Curr. Eye Res. 16: 102-110.


RABBIT ANTI-TIMP-1   (Tissue Inhibitor of Metalloproteinases-1, Third Loop)

 CAT#: RDI-TIMP1abr $469.00/100ug

Packaging: 100ug of peptide affintiy purified anitbody in 0.1ml (1mg/ml) 30 mM Tris pH 7.4, 0.5M NaC1, 0.05% Na azide.

Immunogen: Synthetic peptide based on the human sequence, within the third loop.

Specificity: Binds to TIMP-1. No cross reactivity with other TIMP family members (TIMP-2, TIMP-3). Reactivity with human (also tested on rat C6 cells) Other species not tested.

Applications: -Western Blotting under reducing conditions: identified a band at 30 kDa. This antibody does not bind well to the non-reduced protein, and has limited use for immunoprecipitation, immunohistochemistry and ELISA applications.

TIMP-1 is produced at low (pg/mL) levels by most cell types in culture. Treatment of cells with the phorbol ester TPA stimulates production of TIMP-1 in some cell types, but the low protein levels produced often require concentration of cell culture media to visualize the bands by Western Blotting.

Dilution: starting dilution for Western Blots is 1:1,000 when using colorometric 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


RABBIT ANTI-TIMP-2   (Tissue Inhibitor of Metalloproteinases-2)

CAT#: RDI-TIMP2abr   $469.00/100ug

Packaging: 100ug of peptide affintiy purified anitbody in 0.1ml (1mg/ml) 30 mM Tris pH 7.4, 0.5M NaC1, 0.05% Na azide.

Immunogen: Synthetic peptide based on the carboxy-terminus of the human TIMP-2 sequence.

Specificity: Binds to TIMP-2. No cross reactivity with other TIMP family members (TIMP-1, TIMP-3 or TIMP-4). Reactivity with human (also tested on rat C6 cells) TIMP-2 sequences are well conserved among species, including human,rat, mouse, pig and bovine.

Applications: -Western Blotting under reducing conditions: identified a band at 21 kDa. This antibody does not bind well to the non-reduced protein, and has limited use for immunoprecipitation, immunohistochemistry and ELISA applications.

TIMP-2 is produced at low (pg/mL) levels by most cell types in culture. The low protein levels produced often require its concentration from cell culture media to visualize the bands by western blotting.


Dilution: starting dilution for Western Blots is 1:1,000 when using colorometric 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


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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