rev:   July 7, 2004

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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.


Cardiac Troponin I  (inquire for our troponin brochure)   http://www.researchd.com/troponin/troponindatapack.pdf
see new monoclonal and polyclonals below

$440.00/1mg      $406.00/mg 2-9mg      $375.00/mg 10+ (unless indicated)

-custom Biotin or HRP labeling available on request


NEW MONOCLONAL ANTIBODIES TO HUMAN CARDIAC TROPONIN I

Cardiac Troponin I plays an essential role in diagnosing myocardial damage and is becoming increasingly important as a prognostic marker and in risk stratification of non-ST-elevation ACS patients. This calls for cTnI assays with improved analytical sensitivity and precision. There are many different factors what could influence performance of troponin I assay and affect antibody selection strategy (1). Recently the occurrence of a common interfering factor (IF) that negatively affects determinations of cTnI has been reported (2).

In this Product Release we are describing new monoclonal antibodies to cardiac troponin I which could be suitable for development of interference-free cTnI immunoassay

Reference:

1. Katrukha Alexei "Antibody Selection strategies in cardiac troponin assays" in Cardiac Markers, Humana Press, 2003

2. Susann Eriksson, Miia Junikka, Paivi Laitinen, Kirsi Majamaa-Voltti, Henrik Alfthan, Kim Pettersson "Negative Interference in Cardiac troponin I immunoassays from a frequently occurring serum and plasma component", Clinical Chemistry 49:7, 1095-1104 (2003)

Ordering information:
Product Cat.# Clone Subclass Remarks
Anti-cardiac troponin I RDI-TRK4T21-MF4 MF4 IgG1 EIA, WB
Anti-cardiac troponin I RDI-TRK4T21-M155 M155 IgG1 EIA, WB
Anti-cardiac troponin I RDI-TRK4T21-M18 M18 IgG1 EIA, WB
Anti-cardiac troponin I RDI-TRK4T21-19C7 19C7 IgG2b EIA, WB
Anti-cardiac troponin I RDI-TRK4T21-8E10 8E10 IgG1 EIA, WB
Anti-cardiac troponin I RDI-TRK4T21-16A11 16A11 IgG1 EIA, WB
Anti-cardiac Troponin C RDI-TRK4T27-7B9 7B9 IgG1 EIA, WB

BEST RECOMMENDED PAIRS FOR IMMUNOASSAY (Figure 1)

Coating antibody - Detection antibody :

Note Detection ab would have to be labeled for use in immunoassay, Biotin, HRP or other (not provided labeled unless custom ordered)

M18 (coating) - MF4 (detection)

MF4 (coating) - M155 (detection)

19C7 + M18 (mixture for coating) - MF4 + 8E10 (mixture for detection)

19C7 + M18 (mixture for coating) - MF4+16A11 (mixture for detection)

19C7 + MF4 (mixture for coating) - 7B9 (detection)


Other pairs (coating-labelling)for ELISA:

19C7     -   23C6,

19C7     -     3C7 (best for immunochromatographic assay)

3C7       -     19C7 (for immunochromatographic assays)

23D10   -     23C6

19C7     -     8E10

19C7     -     8E10

19C7     -     16A11

8E10-19C7, 19C7-8E10, 19C7-16A11, 16A11-19C7, 110-111,   110-7F4, 110-mixture of 111 and 7F4, 2A3-10B11.


Available CLONES & Pairing Data   :  Species Reactivity based on Western Blot
Clone subclass epitope human bovine porcine goat canine rabbit cat rat mouse fish
10B11 IgG1 16-APIRR-20 + - + - + + + + + -
4C2 IgG2a 23-SNYRAYA-29 ++ ++ ++ ++ ++ ++ + ++ ++ -
19C7 IgG2b 41-SASRKLQLK-49 ++ ++ + ++ + ++ ++ ++ + ++
414 IgG1 56-AKQELE-61 + + + - + + - - + +
8E10 IgG1 87-LGFAE-91 + + + + + + + - - -
16A11 IgG1 87-LGFAE-91 + + + + + + + - - -
18H7 IgG1 102-RVDKVDE-108 + + + + + + + + + -
7F4 IgG1 190-WRKNIDA-196 + + + + + - + + + -
P4-14G5 IgG1 1-23
P4-3A5 IgG1 1-23
M18 IgG1 13-29
3C7 IgG1 25-40aar ND ND ND ND ND ND ND ND ND ND
M155 IgG1 27-43
16A12 IgG1 85-92aa
16A11 IgG1 87-91aa
M46 IgG1 130-145
3G1 IgG2b 156-171
MF4 IgG1 190-196
C5 IgG1
23C6 IgG2a 15-25aar ND ND ND ND ND ND ND ND ND ND

-all clones except M46 and C5 have no corss reaction with skeletal muscle troponin I

  *Note: clones C5 100% cross with skeletal tropoin I

             clone M46 has approx 5-7 % cross with skeletal troponin Ialso see skeletal troponin I

NOTE:   clone 22B11 recognizes only dephospho-form of cTNi and does not react with mono or biphosphorylated antigen. cat#RDI-TRK4T46  $469.00/mg  or

clone 5E6 (phosphorylated form) cat#RDI-TRK4T45  $469.00/mg


Mouse anti-human Cardiac Troponin I Monoclonals

cat#RDI-TRK4T21-XXXX        XXX=clone number (see below)

$438.00/1mg      $406.00/mg 2-9mg      $375.00/mg 10+

Significant Bulk Discounts Available-please inquire


Custom Biotin labeling : $625.00/1mg  $562.00/mg 2-10  Larger sizes quoted on request, approx 3-4 weeks special order). add-BT to catalog number)
For detection of FREE TROPONIN I:  7F4-414  

For detection of troponin I in complex with troponin C:   use clones 7B9 (coating)-19C7 detection   (7B9=anti-troponin C), 19C7-tropinin I

All antibodies work in western blot and in   immunoaffinity purification.

For WB recommend 16A11,   4G6, 19C7, 6F9 or 7F4.


Some of monoclonal antibodies presented were tested for immunofluorescent staining of TnI in tissue sections and cell cultures. Good results were obtained with MAb 6F9 which can be used for immunodetection of both - cardiac and skeletal TnIs from wide variety of animal species. Other recommended antibody for immunofluorescent staining is clone 19C7.

Immunocytochemistry staining protocol

The cells were fixated 100 % by methanol at - 20C during 20 minutes. After washing with PBS cells were incubated at 10 minutes with solution which blocks nonspecific binding of immunoglobulins. Solution contains 3% of horse serum. Further after three-multiple washing with PBS the cells were incubated with mouse monoclonal antibodies against troponin I with dilution 1: 100 within 12 hours at 4C. After three-multiple washing with PBS the cells were incubated with conjugate (rabbit anti-mouse immunoglobulins) labelled with FITC in dilution 1:200 for 45 minutes at room temperature. After washing the cells with PBS, 60 % solution of glycerol has been added and cells were covered with glass.


Clones: B2, C5:derived from hybridization of Sp2/0 myeloma cells with spleen cells of Balb/c mice immunized with   bovine cardiac troponin I.

CLONEs: 8E10, 19C7, 16A11, 110 (10F4), 111, 112, 10B11, 2A3, 3A1, 5F1, 7F4, 11D7, 414: derived from the hybridization of Sp 2/0 myeloma cells with spleen cells of Balb/c mice immunized with native troponin complex

Clone: 8E10, 190F4, 7F4, 6F9, 414, B2 and C5) derived from free human troponin I

Isotypes: B2, C5, 5F1=mIgG2b.

clones 111, 7F4, 10B11, 2A3, 11D7, 414, 3A1, 8E10=mIgG1.

Clones 110, 112,=mIgG2a

19C7-IgG2b.


19C7=troponin I from human heart. No cross with skeletal muscle troponin I. Clone 19C7 is not affected by heparin,   phosphorylation, oxidation and troponin complex formation.

Cross: B2=troponin I from human, bovine and rabbit heart(). Cross with skeletal muscle troponin I is less than 0.01%. Titer is 10-7.


C5=troponin I from human and bovine heart (also rabbit, mouse, chicken, fish, frog helix heart and skeletal muscle)). Cross reactivity with skeletal muscle troponin I is 5-7%. Titer is 10-6.


110(10B4), 7F4=troponin I from human heart. No cross with skeletal muscle troponin I. Titer >10-7.


111, 10B11, 2A3, 5F1, 11D7=troponin I from human heart. No cross with skeletal muscle                                                          troponin I. Titer >10-6.


112=troponin I from human heart. Some cross with skeletal muscle troponin I is observed. Titer                                                      >10-6


414=troponin I from human heart. Cross with skeletal muscle troponin I is 0.1%. Titer >10-7.



Package: in PBS, pH 7.4 with 0.1% sodium azide as preservative. Store at 4 DEG C.


For In Vitro Research or Further Manufacturing Use Only  Please inquire for other troponin antibodies as well as the pure protein!


Monoclonal mouse anti-cardiac troponin I, phosphorylated form

Catalogue # RDI-TRK4T45   $469.00/mg

Clone: Clone 5E6

This clone has been derived from hybridization of Sp 2/0 myeloma cells with spleen cells of Balb/c mice immunized with phosphorylated form of cardiac troponin I

Specificity: Phosphorylated form of cardiac troponin I.


Mouse isotype: IgG

Application: Antibodies are working in Western blotting and ELISA.

Purification: Purified by chromatography on protein A Sepharose. Purity is tested by electrophoresis.

Presentation: Each vial contains antibodies in (mg/ml ) PBS, pH 7,4, containing 0,1% of sodium azide as preservative. Concentration and lot numbers are indicated on the vial. Store at +4ºC.

Material safety : This product is sold as an antibody preparation for research purpose only. Standard Laboratory Practices should be followed when handling this material. Contains sodium azide (0,1%) as preservative. Although the amount of sodium azide is very small appropriate care must be taken when handling this product.

For Research Use Only


Monoclonal mouse anti-cardiac troponin I, dephosphorylated form

Catalogue # RDI-TRK4T46   $469.00/mg

Clone: Clone 22B11 (20-24 a.a.r.)

This clone has been derived from hybridization of Sp 2/0 myeloma cells with spleen cells of Balb/c mice immunized with dephosphorylated form of cardiac troponin I

Specificity: Dephosphorylated form of cardiac troponin I, no cross-recation with mono or biphosphorylated troponin I.

Mouse isotype: IgG2b

Application: Detection of dephosphorylated cTnI. Antibodies are working in Western blotting and ELISA.
Detection of dephosphorylated cTnI by Western blotting or by sandwich immunoassay cTnI is phosphorylated in vivo and can be phosphorylated in vitro by cAMP-dependent protein kinase A. Sites of phosphorylation are the serines in the 22 nd and 23 d position. It was demonstrated recently that one of these MAbs - namely MAb 22B11 recognizes only dephospho- form of cTnI and does not interact with mono- or biphosphorylated antigen. 22B11 was suggested to be used for qualitative or semi quantitative immunodetection of dephosphorylated cTnI in Western blotting or for quantitative measurements of dephosphorylated cTnI in sandwich immunoassay. Using such immunoassay we recently demonstrated that significant part of cTnI in patient 's blood is partially phosphorylated.

Purification: Purified by chromatography on protein A Sepharose. Purity is tested by electrophoresis.

Presentation: Each vial contains antibodies in PBS, pH 7,4, containing 0,1% of sodium azide as preservative. Concentration and lot numbers are indicated on the vial. Store at +4ºC.

Material safety note: This product is sold as an antibody preparation for research purpose only. Standard Laboratory Practices should be followed when handling this material. Contains sodium azide (0,1%) as preservative. Although the amount of sodium azide is very small appropriate care must be taken when handling this product.



Goat anti-Human Cardiac TROPONIN I   (N terminal affinity purified)


Catalog#: RDI-TROPIabg

Cost: BULK QUOTES ON REQUEST

         $375.00/ 1- 10mgs

         $312.00/mg 11- 49mg

         $250.00/mg 50-100mg

Batch# RDI/tropi/85


Package Size: 1 milligram in 0.2ml (5 mg/ml) in 10mM PBS   pH 7.2 with 0.15 M NaCl and 0.1% sodium azide


Source: goat


Purity: IgG purified using the N-terminal region peptide (27-40)


Specificity: Specific for cardiac Troponin I, not reactive with human skeletal muscle Troponin I by ELISA.


Application -immunoassay

-All titers must be determined in assay

Storage: Store at 4 DEG C(short term)- 20 DEG C long term. Avoid frequent freeze thaw cycles.


Precautions: For In vitro research Use Only. Not for use in or on humans or animals or for diagnostics.


Source: Prepared in the USA in compliance with USDA regulations at a farm registered under NIH, the USDA animal welfare act and USDA veterinary biologics establishment. Export Certificate for animal products available upon request with shipments (allow extra 7 days for delivery due to time delay in obtaining certificates).


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.


Goat anti-Human Cardiac TROPONIN I   (C Region aa 88-106 affinity purified)

Catalog#: RDI-TROPIC88abg

Cost: BULK QUOTES ON REQUEST

$375.00/ 1- 10mgs      $312.00/mg 11- 49mg    $250.00/mg 50-100mg

Source: goat

Purity: IgG purified using the peptide from aa 80

Specificity: Specific for cardiac Troponin I, not reactive with human skeletal muscle Troponin I by ELISA.


Goat anti-human Troponin I (region 14-36)  cat#RDI-TROPI16abG  $375.00/1mg


Rabbit anti-Human Cardiac TROPONIN I

(Coating Antibody)

(affinity purified)


Catalog#: RDI-TROPIabr

Cost: BULK QUOTES ON REQUEST

         $312.00/mg 1- 10mgs

         $219.00/mg 11- 49mg

         $156.00/mg 50-100mg


Batch# see batch sheet

Package Size: 1 milligram


Source: rabbit

Purity: affinity purified by chromatography on human cardiac troponin I immobilized column. Packaged as 1mg in PBS pH 7.4 with 00.1% merthiolate preservative.

Specificity: reacts with troponin I from human heart. Approx 10% cross with skeletal muscle troponin I.

Application -immunoassay:Recommend to use as the coating anibody with monoclonal clone 7F4 (see our troponin monoclonals) to be used as the labelled detecting antibodies. -All titers must be determined in assay


Storage: Store at 4 DEG C.

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.


RDI Divison of Fitzgerald Industries Intl

34 Junction Square Drive

Concord MA 01742-3049

USA

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

fax     (978) 371-2266

EMAIL:antibodies@fitzgerald-fii.com

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