rev:    January 26, 2005

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  ANTIBODIES  

                                                      (anti-Human and others as indicated)
RDI Divison of researchd 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.


PERFORIN  (see 5 abs below) 

-see also antibodies to Granzyme A & Granzyme B


Rat anti-mouse Perforin (clone CB5.4)

cat# RDI-MPERFOR-CB5    $594.00

Presentation: 100ug in 0.1ml (1mg/ml) of purified antibody in PBS with 0.02% sodium azide

Immunogen: recombinant mouse perforin (aa 98-534)

Isotype: Rat IgG2a

Purity: >95% by SDS-PAGE

Specificity: recognizes mouse perforin (epitope region aa402-534)

Reactivity: Mouse, other species not tested

Use: -western blot: approx 1ug/ml, band 66kda on mouse t cell clone B6.1 (neg control NIH.3T3)

      -immunoprecipitation

     -histochemistry(frozen and paraffin sections) 5-10ug/ml

    -Optimal titers must be determined for each use

 Storage: 4 DEG C short term. For long term, store at -20 DEG C. Avoid freeze thaw cycles

-also FITC conjugated: cat#RDI-MPERFOR-CB5FT $625.00/100 test

-immunofluorescence: 1ul of 100ug/ml stock used to stain 200K cells in a sample volume of 50ul.

For In Vitro Research Use Only


Mouse anti-mouse/human Perforin (clone CE2.10)

cat# RDI-MPERFOR-CE2     $594.00

Presentation: 100ug in 0.2ml (0.5mg/ml) of purified antibody in PBS with 0.02% sodium azide

Immunogen: recombinant mouse perforin (aa 98-534)

Isotype: mouse IgG2a

Purity: >95% by SDS-PAGE

Specificity: recognizes mouse perforin (epitope region aa402-534)

Reactivity: Mouse and human, other species not tested

Use: -western blot (mouse ONLY): approx 1ug/ml, band 66kda on mouse t cell clone B6.1 (neg control HEK293)

      -immunoprecipitation (human and mouse)

      -Optimal titers must be determined for each use

Storage: 4 DEG C short term. For long term, store at -20 DEG C. Avoid freeze thaw cycles

For In Vitro Research Use Only


DATA SHEET: Rat anti-mouse Perforin

Catalog#: RDI-MPerforabX    $562.00/vial

Package Size: 100µl of ascites fluid containing 0.1% NaN3.

Species: rat IgG2a

Antigen: recombinant mouse perforin fragment, mPFP1 (172 amino acid residues His189-Cys320). Fragment contains the membrane binding domain and a part of the EGF-type cysteine-rich domain)

Hybridoma Mouse myeloma (P3U/l) X immunized SD rat spleen cells

CLONE: KM585 (also known as P1-8)

Activity: Reacts with mouse perforin, reacts with the common epitope of perforin existing in NK cells and CTL cells. also cross reacts with human perforin

Application: -immunohistochemistry (frozen sections)   (approx 1:50-1:200). Also cell suspensions.

Some users have reported that paraffin embedded tissue staining can be performed.

NOTE: IT IS RECOMMENDED THAT TREATMENT WITH H202 IN WATER BE SUBSTITUTED FOR H202 IN METHANOL.

-ELISA

-Western blot

-flow cytometry (see ref#5,#6)-not recommended,results not consistent

-titers must be determined for each application

Controls: positive: SPB Be or SPB 2.4 cells negative: P815 cells

Storage: Store at 2-8 Deg C (short term), long term aliquot and store -20 DEG C Avoid frequent freeze thaw cycles.

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

References:

-J. Exp Med 172:1877-80

-Internatonal Immunology 7(2):677-84

-J. Exp Med 171:1269-81

-J. Immunology, 155(6):2972-83

-Science:283:1745-1748 (1999)

-Nature 401:76-79 (1999)

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.

1. Nakata, M. et al. (1990). Constitutive expression of pore-forming protein in peripheral blood ã / T cells: implication for their cytotoxic role in vivo. J. Exp.

2. Kawasaki, A. et al. (1990). Perforin, a pore-forming protein detectable by monoclonal antibodies, is a functional marker for killer cells. Internatl. Immunol.

3. Smyth, M. J. et al. (1990). Interleukin 2 induction of ore forming protein gene expression in human peripheral blood CD8+ T cells. J. Exp. Med. 171:

4. Watanabe, H. et al. (1995). Relationships between intermediate TCR cells and NK1.1+ T cells in various immune organs. J. Immunology. 155(6):

5. Opferman, JT. et al.. (1999) Linear differentiation of cytotoxic effectors into memory T lymphocytes.

6. Slifka, MK et al. (1999) Rapid on/off cycling of cytokine production by virus-specific CD8+ cells.Nature 401: 76-79.


Purified Rabbit Anti-rat Perforin (rat/mouse)

cat# RDI-RTPERFORabr   $438.00

Packaging: 200ug Purified rabbit immunoglobulins, lyophilized (Reconstitute to 1 mg/ml by adding 200 ul PBS; contains 0.1% sodium azide)

Immunogen: E. coli-expressed rat perforinUse: 1ug/ml with chemiluminescence detection Western blot detection of rat perforin from kidney on Day 3 in glomerular nephritis model(band approx 66kda)

-This neutralizing polyclonal antibody, which reacts with both rat and mouse perforin, The tested titer for Western blot is 1:1,000. Cross-reactivity to perforin of other species has not been determined.

Storage: 4°C for short term storage or -20°C in small aliquots for long term storage. Avoid repeated freeze and thaw.

Background: Perforin (pore-forming protein) is originally cloned from mouse cytotoxic T-lymphocyte (CTL)1,2, . It is a 555-amino acid protein with a 21-amino acid signal peptide. Perforin is a cytolytic mediator produced by killer lymphocytes, and is stored in and released by cytoplasmic granules. Perforin is involved in the killing mediated by CTL which represents an important mechanism in the immune defense against tumors and virus infections 3,4 .

Reference: 1. Lichtenheld, M.G. et al. (1988) Structure and function of human perforin. Nature 335:448-451 2. Kwon, B.S. et al. (1989) The structure of the mouse lymphocyte pore-forming protein perforin. Biochem Biophys Res Commun 158:1-10 3. Lowin, B. et al. (1992) Perforin and its role in T lymphocyte-mediated cytolysis. Experientia 48:911-920

4. Liu, C.C. et al. (1995) Perforin: structure

For Research Use Only


DATA SHEET:    mouse anti-human Perforin

Catalog#:Catalog#: RDI-Perforabm               $375.00/vial purified

                                RDI-PERFORabm-BT    $469.00/vial Biotin label

                                RDI-PERFORabm-FT    $469.00/vial FITC label

                                RDI-PERFORabm-PE    $594.00/vial PE 120 Tests

Package Size: 100ug purified

                       100ug Biotinylated

                       120 tests FITC labeled or PE labeled

Package Size: 100ug purified- 100ug Biotinylated -   120 tests FITC labeled

Species: mouse IgG2bk

Antigen: purified granules from human YT lymphoma cell line

LOT# see sheet with shipment

CLONE: deltaG9


Activity: Reacts with human perforin of 70kD.


Application: flow cytometry, immunoprecipitation frozen sections,

-titers must be determined for each application


Storage: Store at 4 Deg C. DO NOT FREEZE.


References: Am J Pathol 140:1025-1030

                   Immunol 81:291-295

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.

Bulk Quotes on Request


DATA SHEET: mouse anti-human Perforin:FITC conjugated

Catalog#: RDI-PERFORabm-FT    $469.00/vial FITC label

Package Size: 120 tests FITC labeled in 0.2ml 50mM sodium phosphate pH 7.5, 100mM Potassium Chloride, 150mM NaCl, 5% glycerol, 0.2% BSA and 0.04% NaN3.

Species: mouse IgG2bk

Antigen: purified granules from human YT lymphoma cell line

Source: purified from tissue culture supernatant by Protein A, then conjugated to FITC (further purified by desalting column). Antibody conjugate is approx 0.1mg/ml with a fluorescein/IgG molar ratio of 4.4.

LOT#

CLONE: delta G9 (delta G9)

Activity: Reacts with human perforin of 70kD.

Application: flow cytometry, immunoprecipitation frozen sections,

-titers must be determined for each application

Batch data: Dilution 1:50, use 80-ul/test as below: -Ficoll prepared human peripheral blood lymphocytes were stimulated 1 day at 5 million cells/ml in RPMI 10% FBS media containing 5ug/ml Phytohemagglutinin-P. Cells were then collected and fixed for 30 min with 2% buffered formaldehyde after which they were washed two times. Cells were permeabolized using a buffer containing 0.3% saponin, washed and incubated approx 10 minutes with 20ul human IgG at 250 ug/ml (to block non specific binding) and then incubated 45 minutes on ice with 80ul of antiperforin:FITC at a 1:50 dilution . Cells were washed three times, fixed and analyzed on a BD FACS Calibur. A 21.6% sub population of the cells stained positive with a mean shift of 1.5 log10 fluorescent units when compared to a Mouse IgG2b/FITC negative control at a similar concentration. Binding was blocked when cells were preincubated 10 minutes with 20ul of a 0.5mg/ml anti-perforin antibody. log10 fluorescent units when compared to a mouse IgG2b/Fitc negative control at a similar concentration. Binding was 21% blocked when cells were pre incubated 10 minutes with 20ul of 0.5mg/ml anti-perforin antibody.

Storage: Store at 4 Deg C. DO NOT FREEZE. Protect from light.

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

References: Am J Pathol 140:1025-1030

                    Immunol 81:291-295

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.

Recommended FITC Isotype Control

cat#RDI-MIGGBBFTX2  $312.00/vial 100ug   $250.00/vial when ordered with antibody

Recommended PE Isotype Control

cat#RDI-MIGG2BPEX2 $312.00/vial 100ug

Biotin isotype control: cat#RDI-MIGG2BBTX2   $312.00/100ug

Bulk Quotes on Request


RDI Divison of researchd Industries Intl

San Jose, 95123 CA Snell ave 658

USA

or (800) 370-2222

EMAIL:margaret@cellular-research.com

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