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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, histochemistry, 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.
cat# RDI-PGP95abm $535.00/vial
Background: Protein gene product (PGP) 9.5 is a neuron specific protein,
structurally and immunologically distinct from neuron specific enolase. The
protein which has a molecular weight of 27kD was first identified by high
resolution two dimensional PAGE. Standard immunohistochemical techniques
have demonstrated the presence of PGP9.5 in neurons and nerve fibres al all
levels of the central and peripheral nervous system, in many neuroendocrine
cells, in segments of the renal tubules, in spermatogonia and Leydig cells
of the testis, in ova and in some cells of both the pregnant and non-pregnant
corpus luteum. PGP9.5 is known to be a member of the ubiquitin C terminal
hydroxylase family and is also concentrated with inclusion bodies suggesting
that such structures may be metabolically dynamic regions of the cells.
Specificity:* recognizes human protein gene product 9.5
Presentation: 1 ml lyophilized supernatant with 15mM sodium azide added.
Clone: 10A1
Isotype: mIgG2b
Antigen: Prokaryotic recombinant fusion protein corresponding to the full
length of the protein gene product 9.5 molecule
Hybridoma: mouse myeloma (p3-NS1-Ag4-1)
USES: -acetone fixed frozen sections
-Paraffin sections (using high temp unmasking technique, 0.01M citrate buffer pH 6.0, approx 10 minutes)
-typical dilutions:1:20-1:40, 60 minutes incubation at 25 DEG C with ABC detection
-western blot:1:500-1:1000
Stain: Cytoplasmic and nuclear
Controls: small intestine; enteric ganglion cells
Reconstitution: 1 ml distilled water. Allow to set 30-60 minutes, mix well.
Storage: Store lyophilized material at 4 DEG C. Store reconstituted material
at 4 DEG C for immediate use. For long term use, recommend aliquoting and
store at -20 DEG C. Avoid frequent freeze thaw cycles. Store in non frost
free freezer.
general ref:
-Lowe J et al, Ubiquitin carboxy-terminal hydrolase (PGP 9.5) is selectively present in ubiquitinated inclusion bodies characteristic of human neurodegenerative diseases. journal of pathology 161:153-160 (1990)
-Wilkinson K D et al, The neuron specific protein PGP9.5 is a ubiquitin carboxyl-terminal hydrolase. Science 246:670-675 (1989)
-Wilson POG et al, The immunolocalization of protein gene product 9.5 using rabbit polyclonal and mouse monoclonal antibodies. Br.J.Exp. Path 69:91-104 (1988)
-Day I N M et al, Molecular cloning of cDNA coding for human PGP9.5 protein. A novel cytoplasmic marker for neurons and neuroendocrine cells FEBS Letters 210(2 157:160(1987)
-Thompson RJ et al. PGP9.5-a new marker for vertebrate and neuroendocrine cells. Brain Research 278:224-228 (1983)
FOR IN VITRO RESEARCH USE ONLY-NOT FOR USE IN DIAGNOSTICS
cat# RDI-PGP95MabR $375.00/50ul
RDI-PGP95MabR-1 $812.00/150ul
Presentation: liquid whole serum with 0.05% sodium azide
Immunogen: 175-191 of soluble cytoplasmic protein of human PGP9.5
GASSEDTLLKDAAKVCL
Reactivity: human, mouse, and rat and pig (others not tested)
Use: -western blot: 1:2000 * ( > with chemiluminescence detection)
Histochemistry: approx 1:4000
Optimal dilutions should be determined by user for each application
Storage: Store frozen. Aliquot as undiluted antisera and immediately place at -20 DEG C. At first thaw, tighten cap, centrifuge for 1-2 minutes at 1000 rpm to concentrate material in vial. Stable at least 6 months at -20 DEG C. Avoid repeated freeze/thaw cycles.
Ref: J Comp Neurol 380, 164-74 (1997)
For Research Use Only
Sample Protocol: Antiserum was used on perfusion fixed tissue. Perfusion 1)calcium-free Tyrode's solution 2) paraformaldehyde-picric acid fixative and 3) 10% sucrose in PBS as a cryo-protectant. Desired tissues were stored overnight in 10% sucrose in PBS. Slide mounted tissue sections were processed for indirect immunofluorescence. Slides were incubated with blocking buffer for 1 hour at room temp. Primary antiserum was diluted with blocking buffer to the appropriate working concentration. Blocking buffer was removed and slides were incubated for 18-24 hours at 4 DEG C with primary antiserum. Slides were rinsed 3 times and then incubated with secondary antibodies for 1 hour at room temp. Slides were again rinsed 3 times and coverslipped. Staining was examined using fluorescence microscopy.
-western blot:solubilized human brain extract was examined by SDS-PAGE (4-20% tris-glycine) under reducing conditions. The gel was transferred to a nitrocellulose membrane and blocked overnight at room temp with casein blocking buffer. Following blocking, primary antiserum diluted 1:2000 in 1% BSA,PBS) was added for 4 hours at room temp. Membranes were then washed and incubated with alkaline phosphatase conjugated secondary antibody for 45 minutes at room temp. After washing, the membranes were treated with BCIP/NBT substrate reagents to visualzie immobilized protein.
cat# RDI-PGP95abGP $375.00/50ul
RDI-PGP95abGP-1 $750.00/150ul
Presentation: liquid whole serum with 0.05% sodium azide
Immunogen: Corresponding to residues 175-191 of soluble cytoplasmic protein
of human PGP9.5 GASSEDTLLKDAAKVCL
Reactivity: human, mouse, and rat (others not tested)
Use: Histochemistry: approx 1:500 Optimal dilutions should be determined
by user for each application
Storage: Store frozen. Aliquot as undiluted antisera and immediately place at -20 DEG C. At first thaw, tighten cap, centrifuge for 1-2 minutes at 1000 rpm to concentrate material in vial. Stable at least 6 months at -20 DEG C. Avoid repeated freeze/thaw cycles.
Ref: J Comp Neurol 380, 164-74 (1997)
For Research Use Only
Sample Protocol: Antiserum was used on perfusion fixed tissue.Perfusion 1)calcium-free Tyrode's solution 2) paraformaldehyde-picric acid fixative and 3) 10% sucrose in PBS as a cryoprotectant. Desired tissues were stored overnight in 10% sucrose in PBS.
Slide mounted tissue sections were processed for indirect immunofluorescence. Slides were incubated with blocking buffer for 1 hour at room temp. Primary antiserum was diluted with blocking buffer to the appropriate working concentration. Blocking buffer was removed and slides were incubated for 18-24 hours at 4 DEG C with primary antiserum. Slides were rinsed 3 times and then incubated with secondary antibodies for 1 hour at room temp. Slides were again rinsed 3 times and coverslipped. Staining was examined using fluorescence microscopy.
see anti-guinea pig secondary antibodies at: http://www.researchd.com/rdiabs/2ndabs.htm
RDI Divison of researchd Industries Intl
San Jose, 95123 CA Snell ave 658
USA
or 408-780-0908
EMAIL:margaret@cellular-research.com
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