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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-RB2C4abm   $469.00/100ug

-human reactive

-clone 2C4, mIgG1 isotype

-immunogen: synthetic human phospho-RB(Ser780) peptide, TRPPTLS(P03)PIPHIP which corresponds to amino acids 774-786 of human Rb

-recognizies only the phosphorylation site sequence RPPTLS(P03)PIPHIP. Detects 105kda of human phospho-RB by immunoblot

-WB: approx 1ug/ml with chemiluminescence detection system

-pos control: Human Jurkat, U937, Molt4, 5637 (ATCC HTB-9 RB mRNA negative) and Saos (ATCC HTB-85, mRNA negative).  -no reaction with mouse NIH3T3, WR19L12a, BaF/3

-ref: Cell 81:323(1995, Embo J., 15:7060 (1996), Mol Cells, 5:191(1995)

-cat#RDI-RBSER780abR    $469.00/100ug

-human and mouse reactive

-rabbit IgG

-immunogen: synthetic human phospho-RB(Ser780) peptide, TRPPTLS(P03)PIPHIP which corresponds to amino acids 774-786 of human Rb

-recognizies only the phosphorylation site sequence RPPTLS(P03)PIPHIP. Detects 105kda of human AND mouse phospho-RB by immunoblot  -WB: approx 1ug/ml with chemiluminescence detection system

-pos control: Human U937, MOLt4, non reactive with WERI-RB-1 or 5637.

-Antibody reacts with phosphorylated Ser780 of prb, but does not react with other phosphorylated Ser or THR (Thr5, ser249, thr252, thr356, thr373, ser608, ser612, ser788, ser807, ser811, thr821, thr826) and non phosphorylated ser780 of prb.

-ref: Cell 81:323(1995, Embo J., 15:7060 (1996), Mol Cells, 5:191(1995)

cat #RDI-RBG3245abm   $406.00/vial 100ug

Clone: G3-245 (also known as mab-245 or Mh-RB-02,20)

Isotype: Mouse IgG1

Immunogen: Trp-EpRb fusion protein

Size: 0.1mg in 0.2ml (0.5mg/ml) 50mM Tris-HCL pH 8.0 with 150mM NaCl and 0.09% sodium azide

Specificity: G3-245 recognizes an epitope between amino acids 332-344 (DARLFDHDKTLQ) of the human retinoblastoma protein (pp110- 114 Rb). G3-245 detects human, monkey, rat, mink R and a putative quail Rb. This antibody has also been referred to as Mh-RB-02,20 and mAb-245. A Trp-E-Rb fusion protein was used as immunogen. The antibody is routinely tested by western blot analysis using MOLT-4 human leukemia cell lysate and immunohistochemistry of formalin-fixed, paraffin- embedded human tissue sections. For paraffin-embedded tissue sections, we use standard immunohistochemical staining procedures, including microwave pretreatment with 10 mM citrate buffer prior to staining. Positive Rb staining is typically seen in a variety of human tumors, including subsets of melanomas, colon cancers, stomach cancers, and osteosarcomas. The antibody was purified from ascites by saturated ammonium sulphate precipitation and DEAE chromatography or from hybridoma tissue culture supernatant by affinity chromatography.|

Background: The retinoblastoma gene encodes a nuclear phosphoprotein (Rb or p110Rb) which is expressed in most normal cells of vertebrates and acts as a tumor suppressor gene pro duct (reviewed in 1-4). An underphosphorylated form of Rb is mainly found in resting or fully differentiated cells, whereas the hyperphosphorylated form is present in proliferating cells. Only the underphosphorylated form of Rb binds specifically to viral oncogenes such as SV40 large T, adenoviral EIA and HPV-E7. This interaction may partially contribute to the transforming activity of these viral oncoproteins. Rb also interacts with several cyclins including A, D, and E as well as the transcriptional activator E2F. The importance of these interactions for the biological function of Rb is still being elucidated.

USAGE: Application include western blot analysis (1-2ug/ml), immunoprecipitation (1-2ug/one million cells), flow cy- tometry, gel shift assays, immunofluorescence microscopy of cultured cells (1-5ug/ml), and immunohistochemical staining of   frozen and formalin-fixed, parraffin-embedded tissue sections (0.1-1.0 ig/ml). Rb migrates as multiple closely-spaced bands between approximately 110-116 kD when sized on denaturing polyacrylamide gels (i.e. by SDS-PAGE). The different bands represent different Rb phosphorylation states, the higher molecular weight bands are more highly phosphorylated than the lower molecular weight bands.

The level of phosphorylation is cell cycle dependent, and may also be cell type dependent (not all forms are seen in all cell types that expresss Rb). Gel conditions influence the actual number of bands observed. In cases where optimal band separation is desire, use a 7 to 10% non-gradient long (at least 12 inches) gel.

-blocking peptide cat#RDI-RBG3245-CP $250.00/100ug

-also available:FITC conjugated ab, cat#RDI-RBG3245-FT $562.00/100 Tests

-Western blot control: MOLT-4 cell lysate cat#RDI-MOLT4-AG $344.00/0.5ml

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE

cat#RDI-RBG992abm    $406.00/100ug   mIgG1 isotype

-human reactive

-binds to epitope in N terminal region aa 1-240 of human Rb

-suitable for western blottting (using Molt-4 leukemia cell lysate as positive control)

cat# RDI-RBabm       $375.00/vial 50ug

       RDI-RBamb-1    $625.00/vial 150ug

Background: The retinoblastoma gene product, pRb, is a 110kDa nuclear phosphoprotein which undergoes differential phosphorylation during the cell cycle. During G1 phase, pRb is predominantly in a hypophosphorylated state and it becomes increasingly phosphorylated  throughout the cell cycle until late mitosis when substantial dephosphorylation occurs. A number of cellular proteins are known to interact with the hypophosphorylated pRb including the E2F transcription factor, several cyclins, RBP-1, RBP-2, c-myc, N-myc, and p46. In addition, a number of viral proteins can associate with  pypophosphorylated pRb including adenovirus E1A protein. SV40 large T antigen, and the human papilloma viral protein E7. These viral proteins can alter normal pRb functions by binding to pRb and displacing the normally associated cellular proteins, disrupting the normal tight regulation of cell cycle progression. Rb is well known as a tumor suppressor gene. This gene is either absent or mutated in many human tumors. Retrovirus-mediated gene transfer of the wild-type Rb gene into several neoplastic cell lines (containing Rb mutations) suppresses their tumorgenicity;however,the mechanism of pRb action remains unclear.

isotype: mIgM

package: 50ug or 150ug

antigen: a peptide fragment from aa 714-921 from mouse pRB (at carboxyl end)

USe: Suitable for western blot on human, dog, rat, mouse and chick, using approx 1ug/ml ab (with enhanced chemiluminescence detection). Postive control human fibroblast cell lysate. Major band at Mw 110Kda

-also suitable for immunofluorescence and histochemistry (using an anti-IgM antibody)

Storage: Store at -20 DEG C upon receipt. Avoid frequent freeze thaw cycles. Recommend aliquoting. Avoid   frequent freeze thaw cycles.

Gen REF: Mancini, M.A. et al, 1994 Proc Natl Acad Sci USA 91:418

                 Hinds, P.W. et al. 1992 Cell 70:993

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

-copyright by owner

Mouse anti- RB (Retinoblastoma Protein)

cat #RDI-RBG3245abm   $406.00/vial 100ug

Clone: G3-245 (also known as mab-245 or Mh-RB-02,20)

Isotype: Mouse IgG1

Immunogen: Trp-EpRb fusion protein

Size: 0.1mg in 0.2ml (0.5mg/ml) 50mM Tris-HCL pH 8.0 with 150mM NaCl and 0.09% sodium azide

Specificity: G3-245 recognizes an epitope between amino acids 332-344 (DARLFDHDKTLQ) of the human retinoblastoma protein (pp110- 114 Rb). G3-245 detects human, monkey, rat, mink R and a putative quail Rb. This antibody has also been referred to as Mh-RB-02,20 and mAb-245. A Trp-E-Rb fusion protein was used as immunogen. The antibody is routinely tested by western blot analysis using MOLT-4 human leukemia cell lysate and immunohistochemistry of formalin-fixed, paraffin- embedded human tissue sections. For paraffin-embedded tissue sections, we use standard immunohistochemical staining procedures, including microwave pretreatment with 10 mM citrate buffer prior to staining. Positive Rb staining is typically seen in a variety of human tumors, including subsets of melanomas, colon cancers, stomach cancers, and osteosarcomas. The antibody was purified from ascites by saturated ammonium sulphate precipitation and DEAE chromatography or from hybridoma tissue culture supernatant by affinity chromatography.

Background: The retinoblastoma gene encodes a nuclear phosphoprotein (Rb or p110Rb) which is expressed in most normal cells of vertebrates and acts as a tumor suppressor gene product (reviewed in 1-4). An underphosphorylated form of   Rb is mainly found in resting or fully differentiated cells, whereas the hyperphosphorylated form is present in proliferating cells. Only the underphosphorylated form of Rb binds specifically to viral oncogenes such as SV40 large T, adenoviral EIA and HPV-E7. This interaction may partially contribute to the transforming activity of these viral oncoproteins. Rb also interacts with several cyclins including A, D, and E as well as the transcriptional act- ivator E2F. The importance of these interactions for the biological function of Rb is still being elucidated.

USAGE: Application include western blot analysis (1-2ug/ml), immunoprecipitation (1-2ug/one million cells), flow cy- tometry, gel shift assays, immunofluorescence microscopy of cultured cells (1-5ug/ml), and immunohistochemical staining of frozen and formalin-fixed, parraffin-embedded tissue sections (0.1-1.0 ig/ml). Rb migrates as multiple closely-spaced bands between approximately 110-116 kD when sized on denaturing polyacrylamide gels (i.e. by SDS-PAGE). The different bands represent different Rb phosphorylation states, the higher molecular weight bands are more highly phosphorylated than the lower molecular weight bands.

The level of phosphorylation is cell cycle dependent, and may also be cell type dependent (not all forms are seen in all cell types that expresss Rb). Gel conditions influence the actual number of bands observed. In cases where optimal band separation is desire, use a 7 to 10% non-gradient long (at least 12 inches) gel.

-blocking peptide cat#RDI-RBG3245-CP $250.00/100ug

-also available:FITC conjugated ab, cat#RDI-RBG3245-FT $562.00/100 Tests

-Western blot control: MOLT-4 cell lysate cat#RDI-MOLT4-AG $344.00/0.5ml

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE

USAGE: Application include western blot analysis (2ug/ml), immunoprecipitation (1-2ug/one million cells),

Rb migrates as multiple closely-spaced bands between approximately 110-116 kD when sized on denaturing polyacrylamide gels (i.e. by SDS-PAGE). The different bands represent different Rb phosphorylation states, the higher molecular weight bands are more highly phosphorylated than the lower molecular weight bands.

The level of phosphorylation is cell cycle dependent, and may also be cell type dependent (not all forms are seen in all cell types that expresss Rb). Gel conditions influence the actual number of bands observed. In cases where optimal band separation is desire, use a 7 to 10% non-gradient long (at least 12 inches) gel.

ref: Nature 329:642-645 (1987)

      Annu Rev Cell Biol 10:1029 (1994)

-Western blot control: MOLT-4 cell lysate cat#RDI-MOLT4-AG $344.00/0.5ml

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE

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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