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!----!> On Fri, 13 Dec 96 16:53:31 you ("tom_frey@bdis.com") wrote:
!>Thanks for providing the information below regarding your experience with anti
!>fas-ligand mAbs. However, I had the impression (possibly mistaken) that at
!>least one of these mAbs recognized an intracellular epitope. Could you
clarify
!>which are able to stain cells without premeabilization?
!>
!>Tom_Frey@bdis.com
Dear Mr. Frey,
your impression is absolutely correct. The Fas-ligand mAb clone 33 by
Transduction Labs. stains an intracellular epitope on the fas-ligand
protein. Therefore this antibody can only be used in staining procedures
that (fix and) permeabilize cells prior to antibody incubation. When using
commercial fixation and permeabilization kits like FIX and PERM by An der
Grub (AUSTRIA) this antibody gives very nice staining results.
For any further information about antibodies against fas and fas-ligand,
check the listings of information I gathered below (attached as txt-file on
Windows format; lines exceed 80 chars).
If anything remains to be clarified, do not hesitate to contact me.
Best regards, merry X-MAS, and happy new 1997.
Andreas Christaras
Heinrich-Heine University Children's Hospital Duesseldorf
Lab. of Experimental Hematology (KMT-Labor)
Gebaeude (Bldg.) 14.82
Moorenstrasse 5
40225 Duesseldorf - GERMANY
FON : +49-211-81-16103
FAX : +49-211-81-16191
E-Mail : christar@uni-duesseldorf.de
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FAS and FAS-LIGAND ANTIBODIES as of: 19. Dec 1996
FAS-Antigen (CD95, APO-1) Fas-Ligand (CD95L,=
APO-1L) =09
Product=
Name=
Clone=
Isotyp=
Host Target Presentation Amount Features Epitope application Order=
No. Manufacturer net USD net DEM Price DEM Personal Comments Product=
Name Clone Isotyp Host Target=
Species Presentation Amount Features Epitope Applications Order=
No. Manufacturer Provider net Price USD net Price DEM Price DEM=
Personal Comments
anti-FAS UB2 IgG1 mouse H liquiid purified 100 =B5g none surface FCM,=
frozen section Immunotech 658,00 DM 756,70 DM no information=
available Human Fas-Ligand NOK-1 IgG1 mouse H liquid purified 100=
=B5g neutralizing surface (membrane+soluble FasL) IP, FCM,=
ELISA 65321A Pharmingen 450,00 DM 517,50 DM "NOK-1 gives a bright=
staining on T-cell clones and few B-cell leukemias on flow cytometry;=
functional assays not tested personally"
anti-FAS UB2 IgG1 mouse H FITC conjugated 50 Tests (1=
mL) none surface FCM, frozen section 1506 Immunotech 658,00 DM 756,70=
DM functions perfectly on flow cytometry when used for expression=
determination (own experience) Human=
Fas-Ligand NOK-1 IgG1 mouse H biotinylated 100 Tests (1=
mL) neutralizing surface (membrane+soluble FasL) IP, FCM,=
ELISA 65322X Pharmingen 540,00 DM 621,00 DM "NOK-1 gives a bright=
staining on T-cell clones and few B-cell leukemias on flow cytometry;=
functional assays not tested personally"
anti-FAS UB2 IgG1 mouse H PE conjugated 50 Tests (1 mL) none surface FCM,=
frozen section 1739 Immunotech 688,00 DM 791,20 DM functions=
perfectly on flow cytometry when used for expression determination (own=
experience) Human Fas-Ligand NOK-1 IgG1 mouse H no azide/low endotoxin 250=
=B5g neutralizing surface (membrane+soluble FasL) IP, FCM,=
ELISA 65320C Pharmingen 484,00 DM 556,60 DM "NOK-1 gives a bright=
staining on T-cell clones and few B-cell leukemias on flow cytometry;=
functional assays not tested personally"
anti-FAS ZB4 IgG1 mouse H liquiid purified 100 =B5g inhibits=
apopto surface FCM, culture, frozen 1737 Immunotech 658,00 DM 756,70=
DM "personal experience shows contradicting results; some apoptosis=
induction is inhibited, some not." FAS-Ligand=
(CD95L) 33 IgG1 mouse H lyophilized 150 =B5g cytoplasm (C-terminal aa=
116-277) WB, IF, (FCM) F37720 Transduction Laboratories Dianova 33 gives=
bright and clear staining when cells are sufficiently permeabilized and=
fixed prior to flow cytometric staining
anti-FAS CH11 IgM mouse H liquiid purified 50 =B5g induces=
apoptos surface FCM, WB, cell culture 1504 Immunotech 658,00 DM 756,70=
DM personal experience shows that CH11 functions perfectly as inducer of=
apoptosis FAS-Ligand (CD95L) 33 IgG1 mouse H lyophilized 350 =B5g=
cytoplasm (C-terminal aa 116-277) WB, IF, (FCM) F37720 Transduction=
Laboratories Dianova 33 gives bright and clear staining when cells are=
sufficiently permeabilized and fixed prior to flow cytometric staining
CD95 (FAS/APO-1)DX2 IgG1 k mouse H liquiid purified 100 =B5g induces=
apoptos surface FCM, frozen section 33451A Pharmingen 390,00 DM 448,50=
DM no information available FAS-Ligand (Ab-2) ??? (Ab-2) IgG1=
kappa mouse M (H) lyophilized 100 =B5g cytoplasm (C-terminal aa=
261-277) WB AM-12 Oncogene/Calbiochem Dianova "to my knowledge not=
available in Germany yet; see further remarks (1)"
CD95 (FAS/APO-1)DX2 IgG1 k mouse H FITC conjugated 100 Tests (1 mL)induces=
apoptos surface FCM, frozen section 33454X Pharmingen 500,00 DM 575,00=
DM colleague's experience on T-cells shows a very good staining in flow=
cytometry Human Fas-Ligand NOK-2 IgG2a mouse H no azide/low endotoxin 250=
=B5g neutralizing surface (membrane+soluble FasL) IP, FCM,=
ELISA 65330C Pharmingen 450,00 DM 517,50 DM not tested personally
CD95 (FAS/APO-1)DX2 IgG1 k mouse H PE conjugated 100 Tests (1 mL)induces=
apoptos surface FCM, frozen section 33455X Pharmingen 550,00 DM 632,50=
DM no information available Human Fas-Ligand NOK-2 IgG2a mouse H liquid=
purified 100 =B5g neutralizing surface (membrane+soluble FasL) IP, FCM,=
ELISA 65331A Pharmingen 848,00 DM 975,20 DM not tested personally
CD95 (FAS/APO-1)DX2 IgG1 k mouse H biotinylated 100 Tests (1 mL)induces=
apoptos surface FCM, frozen section 33452X Pharmingen 650,00 DM 747,50=
DM no information available Human Fas-Ligand G247-4 IgG1 mouse H liquid=
purified 100 =B5g surface (membrane+soluble FasL) WB, IP, ELISA,=
FCM 65431A Pharmingen 450,00 DM 517,50 DM not tested personally
CD95 (FAS/APO-1)DX2 IgG1 k mouse H no azide/low endotoxin 500 =B5g induces=
apoptos surface FCM, frozen section 33540D Pharmingen 848,00 DM 975,20=
DM no information available FAS-L (N-20) polyclonal IgG rabbit M, R,=
H liquid purified 100 =B5g surface (N-terminal aa 2-21) WB,=
IC sc-834 Santa Cruz Biotechnologies no information available
FAS CD95 7C11 IgM mouse H RD1 conjugated 50 Tests (1=
mL) none surface FCM Immunotech - DM no information=
available FAS-L (Q-20) polyclonal IgG rabbit H, R, M liquid purified 100=
=B5g surface (N-terminal aa 2-21) WB, IC sc-956 Santa Cruz=
Biotechnologies no information available
anti-FAS UB2 IgG1 mouse H liquiid purified 50 Tests (1=
mL) none surface FCM, frozen section Kamiya 280,00 - DM "no=
personal experience available; if it is functioning like Immunotech's=
version, then same comment applies" FAS-L=
(C-20) polyclonal IgG rabbit H liquid purified 100 =B5g cytoplasm=
(C-terminal aa 260-279) WP, IC sc-957 Santa Cruz Biotechnologies no=
information available
anti-FAS UB2 IgG1 mouse H FITC conjugated 50 Tests (1=
mL) none surface FCM, frozen section Kamiya 290,00 - DM "no=
personal experience available; if it is functioning like Immunotech's=
version, then same comment applies" =09
anti-FAS UB2 IgG1 mouse H PE conjugated 50 Tests (1 mL) none surface FCM,=
frozen section Kamiya 260,00 - DM "no personal experience=
available; if it is functioning like Immunotech's version, then same=
comment applies" =09
anti-FAS ZB4 IgG1 mouse H liquiid purified 100 =B5g inhibits=
apopto surface FCM, culture, frozen Kamiya 260,00 - DM "no=
personal experience available; if it is functioning like Immunotech's=
version, then same comment applies" =09
anti-FAS CH11 IgM mouse H liquiid purified 100 =B5g induces=
apoptos surface FCM, WB, cell culture Kamiya 260,00 - DM no=
information available =09
anti-FAS SM1/1 IgG2a mouse H liquiid purified 50 =B5g induces=
apoptos surface FCM, IF Kamiya 260,00 - DM no information=
available =09
anti-FAS SM1/17 IgG2b mouse H liquiid purified 100 Tests (1=
mL)none surface FCM Kamiya - DM no information=
available =09
anti-FAS SM1/23 IgG2b mouse H liquiid purified 50 =B5g inhibits=
apopto surface FCM Kamiya 195,00 - DM no information=
available =09
anti-FAS SM1/1 IgG2a mouse H liquiid purified 50 =B5g induces=
apoptos surface FCM, IF Bender MedSystems - DM no information=
available =09
anti-FAS SM1/17 IgG2b mouse H liquiid purified 100 Tests (1=
mL)none surface FCM Bender MedSystems - DM no information=
available =09
anti-FAS SM1/23 IgG2b mouse H liquiid purified 50 =B5g inhibits=
apopto surface FCM Bender MedSystems - DM no information=
available =09
human Fas G254-274IgG1 mouse H liquiid purified 100 =B5g=
none surface ELISA capture mAb, WB 65311A Pharmingen - DM no=
information available =09
CD95 (human FAS)B-G27 IgG2a mouse H liquiid purified 200 Tests (2 mL)induces=
apoptos surface FCM CBL527 Cymbus Bioscience 358,00 DM 411,70 DM no=
information available =09
CD95 (human FAS)B-G27 IgG2a mouse H FITC conjugated 200 Tests (2 mL)induces=
apoptos surface FCM CBL527F Cymbus Bioscience 408,00 DM 469,20 DM no=
information available =09
CD95 (human FAS)B-G27 IgG2a mouse H biotinylated 100 =B5g induces=
apoptos surface FCM CBL527B Cymbus Bioscience 408,00 DM 469,20 DM no=
information available =09
CD95 (human FAS)B-G27 IgG2a mouse H no azide/low endotoxin 500 =B5g induces=
apoptos surface FCM, cell culture CBL527N Cymbus Bioscience 844,00 DM =
970,60 DM no information available =09
Fas (Ab-2) DX2 IgG1 k mouse H lyophilized 100 =B5g induces=
apoptos surface FCM, IF AM01 Calbiochem/Oncogene 478,00 DM 549,70 DM=
no information available =09
Fas (Ab-3) DX3 IgG2a k mouse H lyophilized 100 =B5g unknown surface FCM,=
IF AM02 Calbiochem/Oncogene 478,00 DM 549,70 DM no information=
available =09
=09
=09
Fas-Ligand (CD95L, APO-1L) =09
Product Name Clone Isotyp Host Target=
Species=
Presentation=
Amount Features Epitope Applications OrderNo.Manufacturer net USD net=
DEM DEM Personal Comments
Human Fas-Lig NOK-1 IgG1 mouse H liquid purified 100=
=B5g neutralizing surface (membrane+soluble FasL) IP, FCM,=
ELISA 65321A Pharmingen 450,00 DM 517,50 DM NOK-1 gives a bright=
staining on T-cell clones and few B-cell leukemias on flow cytometry;=
functional assays not tested personally
Human Fas-Lig NOK-1 IgG1 mouse H biotinylated 100 Tes neutralizing surface=
(membrane+soluble FasL) IP, FCM, ELISA 65322X Pharmingen 540,00 DM 621,00=
DM NOK-1 gives a bright staining on T-cell clones and few B-cell leukemias=
on flow cytometry; functional assays not tested personally
Human Fas-Lig NOK-1 IgG1 mouse H no azide/low endotoxin 250=
=B5g neutralizing surface (membrane+soluble FasL) IP, FCM,=
ELISA 65320C Pharmingen 484,00 DM 556,60 DM NOK-1 gives a bright=
staining on T-cell clones and few B-cell leukemias on flow cytometry;=
functional assays not tested personally
FAS-Ligand 33 IgG1 mouse H lyophilized 150 =B5g cytoplasm (C-term aa=
116-277) WB, IF, (FCM) F37720 Transduction Laboratori 33 gives bright=
and clear staining when cells are sufficiently permeabilized and fixed=
prior to flow cytometric staining
FAS-Ligand 33 IgG1 mouse H lyophilized 350 =B5g cytoplasm (C-term aa=
116-277) WB, IF, (FCM) F37720 Transduction Laboratori 33 gives bright=
and clear staining when cells are sufficiently permeabilized and fixed=
prior to flow cytometric staining
FAS-Ligand ?(Ab-2) IgG1 k mouse M (H) lyophilized 100 =B5g cytoplasm=
(C-terml aa 261-277) WB AM-12 Oncogene/Calbiochem to my knowledge not=
available in Germany yet; see further remarks (1)
Human Fas-Lig NOK-2 IgG2a mouse H no azide/low endotoxin 250=
=B5g neutralizing surface (membrane+soluble FasL) IP, FCM,=
ELISA 65330C Pharmingen 450,00 DM 517,50 DM not tested personally
Human Fas-Lig NOK-2 IgG2a mouse H liquid purified 100=
=B5g neutralizing surface (membrane+soluble FasL) IP, FCM,=
ELISA 65331A Pharmingen 848,00 DM 975,20 DM not tested personally
Human Fas-Lig G247-4 IgG1 mouse H liquid purified 100 =B5g surface=
(membrane+soluble FasL) WB,IP,ELISA,FCM 65431A Pharmingen 450,00 DM=
517,50 DM not tested personally
FAS-L (N-20) polycl IgG rabbit M, R, H liquid purified 100 =B5g surface=
(N-terminal aa 2-21) WB, IC sc-834 Santa Cruz Biotechnolog no=
information available
FAS-L (Q-20) polycl IgG rabbit H, R, M liquid purified 100 =B5g surface=
(N-terminal aa 2-21) WB, IC sc-956 Santa Cruz Biotechnolog no=
information available
FAS-L (C-20) polycl IgG rabbit H liquid purified 100 =B5g cytoplasm=
(C-terminal aa 260-279) WP, IC sc-957 Santa Cruz Biotechnolog no=
information available
=09
=09
Abbreviations:=09
Applications "FCM: flow cytometry; IF: Immunofluorescence; ICH:=
Immunohistochemistry; WB: Western Blotting; IP:=
Immunoprecipitation" =09
Target Species "H: Human; M: Mouse; R: Rat; MO: Monkey (Simian); RB:=
Rabbit" =09
Remarks: =09
(1) According to information I recieved by product manager of the German=
distributor (Dianova) of Calbiochem/Oncogene apoptosis product series, this=
monoclonal antibody against the fas-ligand is specific for the murine=
protein and N O T (!!!) for human. =09
It is raised against the murine amino acid residues 261-277 that are to be=
found within the cytoplasmic domain. Interestingly, the catalogue indicates=
human reactivity on an other page. =09
=09
GENERAL REMARKS:Prices are indicated only if information about US dollar=
pricing was available to me. Some companies provided me a price list in=
DEUTSCHMARKS, but as my anticipated most (flow) cytometrists do not reside=
in Germany. Therefore anyone interested in=
these =09
antibodies have to inquire for prices not indicated in aforementioned=
listings. All information is either acquired by phone calls to companies=
and/or slaes representatives or is drawn from catalogues available to=
me. =09
No liability or responsebility can be taken for information provided here,=
though I tried to be as precise as I could. =
=09
=09
Because of growing interest in apoptosis, any further information or=
update on these listings are hereby cordially=
welcomed. =09
=09
Commercial providers (manufacturers/distributors) mentioned=
above =09
Kamiya Biomedical 910 Industry Drive, Tukwila, WA 98188,=
USA =09
FON: +1-206-575-8068 =09
FAX: +1-206-575-8094 =09
WEB: NONE =09
=09
Immunotech US residents: inquire Coulter Corp., Hileah, FL,=
USA =09
WEB: http://www.immunotech.com/ =09
=09
Transduction Laborat 2134 Nicholasville Rd., Suite 18, Lexington, KY=
40503, USA =09
FON: +1-606-278-1399 +1-800-227-4063 (both=
orders) =09
FON: +1-606-278-7836 (technical assistance) =
=09
FAX: +1-800-276-2251 (orders) =09
WEB: http://translab.com/ =09
=09
Oncogene 80 Rogers St., Cambridge, MA 02142,=
USA =09
FON: +1-617-492-7289 or +1-800-662-2616 =09
FAX: +1-617-492-8438 or +1-800-828-4871 =09
WEB: http://www.oncogene.com/ =09
E-Mail: resprod@oncogene.com =09
=09
Calbiochem 84 Rogers St., Cambridge, MA 02142,=
USA =09
FON: +1-617-577-9333 or +1-619-450-9600 or +1-800-662-2616 or=
+1-800-662-2616 =09
FAX: +1-800-828-4781 or +1-617-577-8015 =09
WEB: http://www.calbiochem.com/ =09
=09
PharMingen 10975 Torreyana Rd, San Diego, CA 92121,=
USA =09
FON: +1-619-677-7737 or +1-800-848-6227 (customer=
service) =09
FAX: +1-619-677-7749 or +1-800-825-5832 (technical=
service) =09
WEB: http://www.pharmingen.com/ =09
=09
Bender MedSystems Dr. Boehringer Gasse 5-11, A-1121 Vienna,=
AUT =09
FON: +43-1-80105-378 =09
FAX: +43-1-80105-477 =09
=09
Cymbus Biosciences Unit J, Eagle Close, Chandlers Ford, Hants, S053 4NF,=
UK =09
FON: +44-1703-267676 =09
FAX: +44-1703-267677 =09
=09
Santa Cruz Biotechn 2161 Delaware Ave., Santa Cruz, CA 95060,=
USA =09
FON: +1-408-457-3800 or +1-800-457-3801 =09
FAX: +1-408-457-3801 =09
WEB: http://www.scbt.com/ =09
E-Mail: scbt@netcom.com =09
=09
=09
=09
=09
Andreas Christaras
Heinrich-Heine University Children's Hospital Duesseldorf
Lab. of Experimental Hematology (KMT-Labor)
Gebaeude (Bldg.) 14.82
Moorenstrasse 5
40225 Duesseldorf - GERMANY
FON : +49-211-81-16103
FAX : +49-211-81-16191
E-Mail : christar@uni-duesseldorf.de
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web