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RBR1 | Retinoblastoma related protein

345 €

AS11 1627   | clonality: polyclonal  |  host: chicken  |  reactivity: Arabiodpis thaliana, Medicago sativa

PRODUCT INFORMATION IN PDF

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Item No:
AS11 1627

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product information
Background

The retinoblastoma protein Rb is considered to be a key regulator of G1/S phase transition by blocking S phase entry and cell growth. Plant retinoblastoma-related (RBR) proteins share a homology with the human tumour suppressor retinoblastoma (pRb) protein. RBR protein functions are controlled by phosphorylation and protein-protein interactions. Short name: AtRBR

Immunogen

Recombinant C-terminal fragment consisting of 236 amino acids  of Arabidopsis thaliana retinoblastoma protein Q9LKZ3, locus At3g12280

Host Chicken
Clonality Polyclonal
Clone
Purity Total IgY
Format Liquid in PBS, pH 8.5 with 0.02 % sodium azide
Quantity 2 x 50 µl
Reconstitution
Storage

Store at 4°C. Upon arrival please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tubes.

Tested applications
Immunoprecipitation (IP), Western Blot (WB)
Related products
Additional information

Total IgY concentration is 30.8 mg/ml

application information
Recommended dilution

1 : 2000 (WB), 2µl (IP)

Expected | apparent MW

112 kDa

Confirmed reactivity Arabidopsis thaliana, Medicago sativa
Predicted reactivity
dicots: Camelia sinensis, Chenopodium rubrum, Pisum sativum, Scutelaria baicalensis, monocots: Cocos nucifera, Hordeum vulgare, Zea mays, Oryza sativa, trees: Populus tremula
Not reactive in

Chlamydomonas reinhardtii

Additional information

Antibody is not suitable for immunolocalization.

Methanol concentration in a transfer buffer can be considerably reduced or for a better transfer of high MW proteins (even with PVDF membrane).

For immunoprecipitation start with 2 µl and titrate it depending upon your experimental conditions. Please note that you work with a total IgY fraction, which means that it will contain between 40-60 µg of total IgY (directed not only against retinoblastoma) therefore all of this IgY needs to be captured by the anti-IgY matrix.

As control pre-serum for IP this product can be used, total, pre-immune IgY

Selected references Cheng et al. (2013). Down-regulation of multiple CDK inhibitor ICK/KRP genes up-regulates E2F pathway and increases cell proliferation, organ and seed sizes in Arabidopsis. Plant j. May 7.
Ábrahám et al. (2011). Immunodetection of retinoblastoma-related protein and its phosphorylated form in interphase and mitotic alfalfa cells. J Exp Bot 62(6):2155-2168.

Application example

western blot detection of plant retinoblastoma protein
35 µg of total proteins from Arabidopsis thaliana Col-0 cell suspension culture (1) or one week old seedlings (2) as well as 3.5ng of purified GST-RBR1 fusion protein (3) were separated on 8% Laemmli SDS polyacrylamide gels and blotted onto PVDF membrane overnight. Filters were blocked in 5% milk powder in TBS-0.05% Tween 20 (TBS-T) for 2 hours then probed with anti-RBR1 antibody (1:6000,  2 hours at RT) and HRP-conjugated rabbit anti-chicken IgY (PIERCE) secondary antibody (1: 20000, 1 hour at RT) in TBS-T containing 5% milk powder. After each antibody incubation steps filters were washed with TBS-T, TBS-T containing 2% milk powder, TBS-T for 10 min each on a rocking platform. Signal was developed with Super Signal West Pico chemiluminescent substrate (PIERCE) and visualized by exposing to a film (Agfa Cronex 5) for 5 min.

Courtesy Dr. Laszlo Bako, Umeå Plant Science Center, Sweden

||| For other applications, usage on species other than stated above or any other questions, please use the LiveChat option or contact us at support@agrisera.com