p130 Antikörper

Produktnummer ABIN487489

Produktdetails anti-p130 Antikörper

Target: p130 (RBL2) (Retinoblastoma-Like 2 (p130) (RBL2))

Reaktivität: Human

Wirt: Maus

Klonalität: Monoklonal

Konjugat: Dieser p130 Antikörper ist unkonjugiert

Applikation: Western Blotting (WB), Immunofluorescence (IF), Immunoprecipitation (IP)

Spezifität: This antibody reacts with RB2.

Kreuzreaktivität (Details): Species reactivity (tested):Human.

Produktmerkmale: Synonyms: Retinoblastoma-like protein 2, RB2, 130 kDa retinoblastoma-associated protein, p130,Retinoblastoma-related protein 2, RBR-2, pRb2

Aufreinigung: Protein-A Sepharose Chromatography.

Immunogen: A peptide from RB2 proteinAA Sequence: KRKRRNSGSSDSRSHQNSPTELNKDRTSRDSSPVMR.

Klon: DCS-211

Isotyp: IgG1

Anwendungsinformationen

Applikationshinweise: Immunoprecipitation/Western Blot: 5 μg/mL. In Western blot, this monoclonal antibody should be used combination withImmunoprecipitation and is not recommended in Western blot using crude cell lysate. Positive Control: Jurkat. Detailed procedure is provided in Protocols.
Other applications not tested.
Optimal dilutions are dependent on conditions and should be determined by the user.

Protokoll: Immunoprecipitation / SDS PAGE & Western Blotting1) Wash the cells 3 times with PBS and suspend with 10 volume of cold Lysis buffer (50 mMTris-HCl, pH 7. 2, 250 mM NaCl, 0. 1% NP-40, 2 mM EDTA, 10% glycerol) containingappropriate protease inhibitors. Incubate it at 4°C with rotating for 30 minutes, thensonicate briefly (up to 10 seconds). 2) Centrifuge the tube at 12,000 x g for 10 minutes at 4°C and transfer the supernatant toanother tube. 3) Add 3 µL of the anti-RB2 monoclonal antibody into 200 µL of the supernatant. Mix welland incubate with gentle agitation for 30-120 minutes at 4°C. Add 20 µL of 50% ProteinA-agarose beads resuspended in the Lysis buffer. Mix well and incubate with gentleagitation for 60 minutes at 4°C. 4) Wash the beads 3-5 times with ice-cold Lysis buffer (centrifuge the tube at 2,500 x g for10 seconds). 5) Resuspend the beads in 20 µL of Laemmli's sample buffer, boil for 3-5 minutes, andcentrifuge for 5 minutes. Load 10 µL of the sample per lane in a 1 mm thickSDS-polyacrylamide gel for electrophoresis. 6) Blot the protein to a polyvinylidene difluoride (PVDF) membrane at 1 mA/cm2 for 1 hourin a semi-dry transfer system. (Transfer Buffer: 25 mM Tris, 190 mM glycine, 20% MeOH). See the manufacture's manual for precise transfer procedure. 7) To reduce nonspecific binding, soak the membrane in 10% skimmed milk (in PBS, pH7. 2) for 1 hour at room temperature, or overnight at 4°C. 8) Incubate the membrane with the anti-RB2 monoclonal antibody (1-5 µg/mL) diluted with1% skimmed milk (in PBS, pH 7. 2) for 1 hour at room temperature. 9) Wash the membrane with PBS (5 minutes x 6 times). 10) Incubate the membrane with the 1: 10000 POD-conjugated anti-mouse IgG diluted with1% skimmed milk (in PBS, pH 7. 2) for 1 hour at room temperature. 11) Wash the membrane with PBS (5 minutes x 6 times). 12) Wipe excess buffer on the membrane, then incubate it with appropriatechemiluminescence reagent for 1 minute. Remove extra reagent from the membrane bydabbing with paper towel, and seal it in plastic wrap. 13) Expose to an X-ray film in a dark room for 5 minutes. Develop the film as usual. Thecondition for exposure and development may vary. Positive Control for Immunoprecipitation/Western blotting: Jurkat

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Konzentration: 1.0 mg/mL

Buffer: PBS, pH 7.2 containing 50 % Glycerol without preservatives.

Konservierungsmittel: Without preservative

Lagerung: -20 °C

Informationen zur Lagerung: Store the antibody (in aliquots) at -20 °C. Avoid repeated freezing and thawing.
Shelf life: one year from despatch.

Haltbarkeit: 12 months

Details zu p130

Target: p130 (RBL2) (Retinoblastoma-Like 2 (p130) (RBL2))

Andere Bezeichnung: RBL2 / p130 (RBL2 Produkte)

Synonyme: RBL2 antikoerper, RBR2 antikoerper, RRB2a antikoerper, PRB2 antikoerper, RBR-2 antikoerper, Rb2 antikoerper, p130 antikoerper, P130 antikoerper, retinoblastoma-like 2 (p130) antikoerper, RB transcriptional corepressor like 2 antikoerper, related to retinoblastoma 2 antikoerper, RBL2 antikoerper, LOC541652 antikoerper, Rbl2 antikoerper

Hintergrund: The product of the Retinoblastoma related human gene RB2 (p130) is highly homologous to the product of the RB tumor suppressor gene, pRB (p105) and the related p107 protein, especially in the functional pocket region of the protein that associates with the E2F transcription factor. Overexpression of RB2 results in a drastic reduction in cell proliferation. Additionally, certain carcinoma-derived cell lines and lung neoplasms show low levels of RB2 expression, possibly due to gene rearrangement, which may suggest a tumor suppressor role for this protein in the pathogenesis and progression of lung cancer.Synonyms: 130 kDa retinoblastoma-associated protein, RB2, RBR-2, Retinoblastoma-like protein 2, Retinoblastoma-related protein 2, p130, pRb2

Gen-ID: 5934

UniProt: Q08999

Pathways: Zellzyklus, Mitotic G1-G1/S Phases