rev: 5/19/2008
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ANTIBODIES
(anti-Human and others as indicated)
RDI Division 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.
anti-CPP32 (CASPASE-3) Antibodies
Rabbit (monoclonal) anti-caspase-3 Active form (human/mouse)
cat#RDI-CASP3ACTabRm2 $451.00/50ug in PBS with 5% trehalose
Package: 1mg in 1 ml PBS 5% trehalose
Specificity: The antibody specifically recognizes the active form of caspase-3 in human and mouse cells. It detects the p17 subunitt of Caspase 3 but does not detect the precursor form.
Use:
-histochemistry: 0.1-0.5 ug.ml
Optimal dilution should be determined by the researcher.
Background: The caspase family of cysteine proteases plays a key role in apoptosis and inflammation (reviewed in 1). Caspase-3 (CPP32, Yama, apopain) is a key protease that is activated during the early stages of apoptosis and, like other members of the caspase family, is synthesized as an in- active proenzyme that is processed in cells undergoing apoptosis by self-proteolysis and/or cleavage by another protease. The processed forms of caspases consist of large (17-22 kD) and small (10-12kD) subunits which associate to form an active enzyme. Active caspase-3, a marker for cells undergoing apoptosis, consists of a heterodimer of 17 and 12 kD subunits which is derived from the 32 kD proenzyme.2 Active caspase-3 proteolytically cleaves and activates other caspases, as well as relevant targets in the cyto- plasm, e.g.,D4-GDI and Bcl-2(3). and in the nucleus, e.g. PARP.
For Research Use Only
rabbit anti-caspase-3-active form (human)
cat#RDI-CASP3ACTabr $406.00/100ug in 0.2ml
cat#RDI-CASP3ACTab-BT Biotin labeled $406.00/100ug
cat#RDI-CASP3ACTab-PE PE conjugated $469.00/100T
-rabbit polyclonal
-antigen: active human caspase-3 fragment
-antibody recognizes a conformational epitope which is exposed by activation-induced cleavage or denaturation of the inactive enzyme.
-at optimal concentrations only sees active caspase-3 (approx 50 fold preference for active form compared to the inactive form). At higher concentrations may cross with inactive form.
-use: histochemistry (acetone fixed frozen sections, cytopsins), flow cytometry. In formalin fixed paraffin sections and western blot will recognize both inactive and active form. In immunoprecipitation, preferentially recognizes active caspase-3 but may also see native form at high concentrations.
Mouse anti-CPP32/caspase-3 (Human)
cat #RDI-CPP32abm $438.00/50ug
#RDI-CPP32abm-1 $750.00/150ug
RDI-CPP32abm-RP $500.00/vial 50ug (HRP conjugated)
RDI-CPP32abm-RPX $812.00/vial 150ug (HRP conjugated)
-suitable for use on western blot (approx 0.5ug/ml), immunoprecipitation (native or denatured conditions), reactive with human .Blots at 32Kda.
-antigen: fragment from human CPP32 1-219 (p20 region)
-mouse IgG2a, packaged as 50ug (200ul) or 150ug (600ul) (at 0.25mg/ml in 50%
glycerol, PBS + 1.5mM NaN3 & 1mg/ml BSA)
For In Vitro Research Use Only
Mouse anti-CPP32 (Caspase-3) Monoclonal (Mouse/Rat)
cat# RDI-CPP32abm-46 $438.00/vial 50ug
RDI-CPP32abm-46X $750.00/vial 150ug
clone ref# 46
isotype: mIgG1
package: 50ug (200ul) or 150ug (600ul) (at 0.25mg/ml in 50% glycerol, PBS +
1.5mM NaN3 & 1mg/ml BSA)
antigen: a protein fragment corresponding to amino acids 25-145 from mouse
Caspase-3.
Use: Suitable for western blot on mouse (rat expected but not verified) using
approx 0.25-0.5ug/ml on total cell lysate of RSV-3T3 (positive control).
Major band =Mw 32 kDa. Also suitable for
immunofluorescence (tested on RSV-3T3 cells).
Storage: Store at -20 DEG C upon receipt. . Recommend aliquoting. Avoid frequent warming cycles.
-copyright by owner
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.
cat#RDI-RSV3T3/CPX $250.00/500ug vial
Rat Caspase 3 (Recombinant)
cat#RDI-TRK8C1 $688.00/5ug vial $625.00/vial 2+ Bulk on request
Application: Caspase-3 (also known as CPP32, Yama, or Apopain) is a member of the cysteine proteases family. Similar as other caspases, caspase-3 also exists in cells as an inactive proenzyme. During the initiation of apoptosis the proenzyme is processed at aspartate residues to form active caspase-3. Involved in the activation cascade of caspases responsible for apoptosis execution. At the onset of apoptosis it proteolytically cleaves poly(ADP-ribose) polymerase (PARP) at a 216-asp-|-gly-217 bond. Cleaves and activates sterol regulatory element binding proteins (SREBPS) between the basic helix-loop- helix leucine zipper domain and the membrane attachment domain. Cleaves and activates caspase-6, -7 and -9. Cleaves DFF45 and thus activates DFF40.
In combination with caspase 3 activity assays, the active recombinant caspase 3 is useful in biological screening of caspase substrates and inhibitors. The recombinant enzyme can also be used as a positive control in caspase 3 assays.
Source: The rat Caspase 3 cDNA was synthesized using mRNA from rat brain. The purified active recombinant rat caspase 3 with C-terminal 6xHis Tag was expressed in E. coli. Heterodimer of a 17 kDa (p17) and a 12 kDa (p12) subunit.
Presentation: µg of protein in 200µl of storage buffer containing 50% glycerol.
Liquid at -20ºC.
Approximately, 180,000 Units of activity per 1mg of protein. One unit of the recombinant caspase 3 is the enzyme activity that cleaves 1 nmol of the caspase substrate z-DEVD-AMC (AMC: aminomethylcoumarin) per hour at room temperature (+20ºC) at 20 µM substrate concentration
Analysis: Purity more 98% by SDS-PAGE.Immunological activity confirmed by reaction with monoclonal antibody that is specific for the rat caspase-3 in immunoblotting.
The protein concentration was determined using the
Storage: At -20 0 C in liquid form. For long storage -70ºC is recommended.
Mouse anti-CPP32 (Caspase-3) clone JMH62 (paraffin)
cat# RDI-CPP32abm-62 $594.00/vial
Package: 1ml lyophilized tissue culture supernatant containing 15mM sodium
azide. Reconstitute with 1ml of sterile water , let
set at least 30 minutes 4 DEG C, Mx well before using.
Clone: JMH62
Isotype: mIgG1
Immunogen: Prokaryotic recombinant protein corresponding to a 182 amino acid
fragment of the CPP32 molecule (caspase-3).
Hybridoma: Mouse myeloma (p3-NS1-Ag4-1).
Reactivity: Cysteine protease protein(CPP)-32. a member of the interleukin-1B-converting enzyme (ICE)
family of mammalian proteases.
Use: Effective on paraffin was embedded tissue using the EDTA pH8.0 high
temperature antigen unmasking technique:
(not recommended for frozen sections)
Typical working dilution 1:25-1:50.
High temperature antigen unmasking using technique 60 minutes primary
antibody incubation at 25"C. Standard ABC technique.
Western blotting: typical working dilution: 1:100
Positive controls: Tonsil
Staining pattern: Cytoplasmic
Storage: Store unopened lyophilized antibody at 1'C. Under these conditions,
there is no significant loss in product performance up to the expiry date
indicated on the vial label The reconstituted antibody is stable for at lease
two months when store at 4'C. For long term storage, it is recommended that
aliquots of the antibody are frozen at -20'C (frost-free freezers are not
recommended). Repeated freezing and thawing must be avoided. Prepare working
dilutions on the day of use.
FOR RESEARCH USE ONLY-Not for use in diagnostic procedures
Background:
Cysteine protease protein (CPP)-32 is a member of the interleukin-1
beta-converting enzyme (ICE) family of mammalian proteases which specifically
cleaves substrates at the C-terminal side of aspartic acid residues. Members of
this family have been implicated in apoptosis and CPP32 (caspase-3) is thought
to act as a control mediator of programmed cell death in mammalian cells. CPP32
is synthesised as an inactive 32kD proenzyme and is processed during apoptosis
to its active form which is responsible for the cleavage of poly (ADP-ribose)
polymerase (PARP), actin and sterol regulatory element binding protein (SREBP) during apoptosis. CPP32 is detected
immunohistochemically in epithelial cells of skin, renal proximal tubules and collecting
ducts, epilhmorlicular cells of the thymus and bronchial, colonic and salivary
duel epithelia. Chondrocytes, bone osteocytes, megakaryocyles, mature neutrophils of bone marrow and plasma cells of
the tonsil, lymph node and bone marrow also stain intensely, This antibody
stains the majority of intermediate to high grade non- Hodgkin's B cell
lymphomas, mantle cell lymphomas, Burkiti's lymphomas and immunoblastic
lympohomas.
References:
Krajewska M. Wang H-G. Krajewski S. et al., Immunohistochemical analaysis of
in vivo patterns of expression of CPP32 (Caspase-3),a
cell death protease. Cancer Research 57: 1605-1613 (1997).
Nicholson D.W. Ambereen A. Thomberry NA. et al,
Identification and inhibition of the ICE-CED-3 protease necessary for mammalian
apoptosis, Nature 376:37-43 (1995).
Fernandes-Alhemri T, Lrtwack g and Alriemri E S CPP32, a novel human apoptotic protein with homology to Casenorhehdotos elegans. cell death protein Ccd-3 end mammalian interleukin-1B converting enzyme. The Journal of Biological chemistry 269(49)-30761-30764 (1994).
-See also recombinant active caspase-3 cat#RDI-CASPASE3-AG $500.00/5ug $750.00/10ug
DATA SHEET: mouse anti-human CD95 clone DX2 (activation use)
no azide formulation: cat#RDI-CD95-DX21X $688.00/0.5mg
Prep: purified from tissue culture supernatant via Protein G affinity
chromatography.
Species: mouse IgG1k
CLONE: DX2
Activity: Reacts with the 45kD transmembrane cell surface FAS or APO-1 antigen,
designated CD95. CD95 is expressed on a variety of normal and neoplastic cells,
and is a polypeptide which plays a role in the programmed sequence of events
leading to cell death (apoptosis). the DX2 clone
specifically reacts with murine L cells, murine L1210 leukemia cells and murine
P815 mastocytoma cells transfected with human Fas cDNA but not with
untransfected parental cell lines. Cross linking with DX2 delivers an apoptotic
signal indicating that DX2 recognizes a functional epitope of CD95 antigen.
Application: -Histochemistry for staining acetone fixed, frozen tissue sections.
-flow cytometry-Titer for each application
References:
1) Schlossman, S. et al, ed.s 1995, Leucocyte Typing V, White Cell Differentiation Antigens, Oxford University Press, NY.
2) J. Exp. Med, 177:1547
3) Cell 66:233
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.
Rabbit Anti-Caspase-3-Active Form (CPP32) MONOCLONAL
cat# RDI-CASP3ACTabRm $406.00/vial
Package: 25ug in 0.050ml PBS pH7.2 with 0.09% sodium azide
antigen: active human caspase-3 fragment
Clone: C92-605
Specificity: The C92-605 monoclonal antibody (mAb) specifically recognizes the
active form of caspase-3 in human and mouse cells It
does not recognize the proenzyme form in any assay so far tested. The mAb was
originally characterized on human and mouse cells by immunohistochemical
staining of apoptotic and non-apoptotic populations and shown to only recognize
cells or tissues induced to undergo apoptosis.
Use: -flow cytometry: approx 0.125-0.5ug/million cells ,using Jurkat T cells
treated with 4uM camptothecin for 5 hours to induce apoptosis,cells were then
fixed and permeabilzied and stained with approx 0.25ug/million cells for 20 min
at room temp in dark. Cells were then washed permeabilzation buffer, stained
with goat anti-rabbit FITC cat#RDI-711096152. approx
50% of treated cels were positve for caspase-3 staining.
-histochemistry: acetone fixed frozen sections approx 0.25-1ug/ml. controsl include untreated Jurkat T cells and treated cells with 6uM camptothecin for 4 hours, ABC/Dab detection -for mice, untreated or treated with 100ug anti-Fas Mab (cat#RDI-MCD95-XP $625.00/0.5mg, clone J02) injected i.p. to induce apoptosis, sacrificed after 6 hours. Cytospins of JurKat cells or frozen tissue sections of mouse liver were acetone fixed and stained with antibody followed by ABC/Dab detection.
-immunoprecipitation: approx 1ug/5 million Jurkat T cells,
must titer to optimize which form is Immunoprecipitated). (treated cells with 6uM camptothecin for 5 hours to induce
apoptosis.
Background: The caspase family of cysteine proteases plays a key role in
apoptosis and inflammation (reviewed in 1). Caspase-3 (CPP32, Yama, apopain) is
a key protease that is activated during the early stages of apoptosis and, like
other members of the caspase family, is synthesized as an in- active proenzyme
that is processed in cells undergoing apoptosis by self-proteolysis and/or
cleavage by another protease. The processed forms of caspases consist of large
(17-22 kD) and small (10-12kD) subunits which
associate to form an active enzyme. Active caspase-3, a marker for cells
undergoing apoptosis, consists of a heterodimer of 17 and 12 kD subunits which
is derived from the 32 kD proenzyme.2 Active caspase-3 proteolytically cleaves
and activates other caspases, as well as relevant targets in the cytoplasm,
e.g.,D4-GDI and Bcl-2(3). and in the nucleus, e.g.
PARP.
ref:
For Research Use Only
Active Recombinant Human Caspase-3 (CPP32)
CAT#: RDI-CASPASE3-AG $500.00/5ug
RDI-CASPASE3-AG1 $750.00/10ug
Packaging: vial of 5ug enzyme in 25 ul in 50 mM Tris, pH 8.0,with 100 mM NaC1, 50 mM imidazole.
Source: E.coli.(When expressed in E. coli, caspase-3,
spontaneously undergoes autoprocessing to yield the subunits characteristic of
the active enzyme).
Activity: The rate of caspase enzymatic hydrolysis was measured by release of AMC from the Ac-DEVD-AMC caspase substrate as emission at 440 nm upon excitation at 380 nm using a spectrofluorometer.
USAGE: The active enzyme is designed to be used in caspase assays.
Storage: Store the enzyme at -80'C. The thawed active enzyme is stable at 4'C
for at least a week. Avoid multiple freeze-thaw cycles and exposure to frequent
temperature changes, which can greatly alter product stability.
BACKGROUND
Caspases are cysteine proteases that play a centrl role in apoptosis (reviewed in 1 and 2). The caspace family was
discovered by searching human cDNA libraries for
sequences homologous to ced-3, aC. elegans death gene
that is required for normal apoptosis during development. The first mammalian
homolog of ced-3 to be identified was ICE(interleukin-1B-converting
enzyme). Subsequent numerous human ced-3 homologues were repidly identified
which led to multiple names for many of the molecules. To achieve
consistency,"caspase" was adopted as a root name for all family
members. The name was selected based on two catalytic properties of these
enzymes, the "c" reflects a cysteine protease mechanism and
"aspase" refers to their unique ability to cleave after aspartic
acid. There are at least 10 members, caspase- 1(ICE), caspace-2 (ICH-1), caspase-3
(CPP32), Yama, apopain), caspase-4 (TX-ICH-2, ICErel-II), caspase-5
(ICErel-III), caspase- 6 (Mch2), caspase-7(Mch3, ICE-LAP3, CMH-1), caspase-8
(MACH,FLICE, Mch5), caspase-9 (ICE-LAP6, Mch6), and caspase- 10(Mch4). Each
caspase is synthesized as an inactive proenzyme that is processed by cleavage
at asparte residues by another protease or by self-proteolysis. The processed
forms consist of large (17-22 kD) and small (10-12 kD) subunits which assocate to form an active enzyme. The activation of some of
these caspases has been shown to occur during apoptosis.
Caspase-3,-6,-7, and -8 have been whown to play a role in apoptosis induced by
the death receptors Fas and tumor necrosis factor receptor type 1 (TNFR1).One
of their substrates is poly (ADP ribose) polynrase (PARP). PARP is an enzyme
that is involved in DNA repair and genomic maintenance. Activated caspase 3, 6,7 and 8 can all clleave PARP from its 116 kD form to an 85
kD residual fragment. The cleavage separates the DNA-binding domain in the
N-terminus of PARP from its C-terminus catalytic domain, and results in loss of
normal PARP function. The cleavage site in PARP is C-terminal to Asp-216. The
upstream sequence of the PARP cleavage site, DEVD (Asp-Glu-Val-Asp), is
utilized as a basis for highly specific caspase-3 substrates such as Ac(N-acetyl)- DEVD-AFC
(7-amino-4-trifluoromethylcourmarin)and Ac(N-acetyl)- DEVD-AMC
(7-amino-4-methylcoumarin) as well as the caspase-3 aldehyde inhibitor
Ac-DEVD-CHO.
For In Vitro Research Use Only
see also antigens for other caspases (caspase-6, caspase-7, caspase-8)
Return (alphabetical
antibody .index)
RDI Division of Fitzgerald Industries Intl
phone (978) 371-6446 or (800) 370-2222
fax (978) 371-2266
EMAIL:antibodies@fitzgerald-fii.com
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