p130 antibody

Catalog No. ABIN487489

Product Details anti-p130 Antibody

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

Reactivity: Human

Host: Mouse

Clonality: Monoclonal

Conjugate: This p130 antibody is un-conjugated

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

Specificity: This antibody reacts with RB2.

Cross-Reactivity (Details): Species reactivity (tested):Human.

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

Purification: Protein-A Sepharose Chromatography.

Immunogen: A peptide from RB2 proteinAA Sequence: KRKRRNSGSSDSRSHQNSPTELNKDRTSRDSSPVMR.

Clone: DCS-211

Isotype: IgG1

Application Details

Application Notes: 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.

Protocol: 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

Restrictions: For Research Use only

Handling

Concentration: 1.0 mg/mL

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

Preservative: Without preservative

Storage: -20 °C

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

Expiry Date: 12 months

Target Details for p130

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

Alternative Name: RBL2 / p130 (RBL2 Products)

Background: 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

Gene ID: 5934

UniProt: Q08999

Pathways: Cell Division Cycle, Mitotic G1-G1/S Phases