RBR1 | Retinoblastoma related protein

AS11 1627 | Clonality: Polyclonal | Host: Chicken | Reactivity: Arabiodpis thaliana, Medicago sativa

Product Information

Immunogen

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

Host Chicken

Clonality Polyclonal

Purity Total IgY

Format Liquid in PBS, pH 8.5 with 0.02 % sodium azide

Quantity 100 µl

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 Chromatin IP (ChIP), Immunoprecipitation (IP), Western blot (WB)

Recommended dilution 2µl (IP), 1 : 2000 (WB)

Expected | apparent MW

112 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana, Medicago sativa

Predicted reactivity Camelia sinensis, Chenopodium rubrum, Cocos nucifera, Hordeum vulgare, Oryza sativa, Pisum sativum, Populus tremula, Scutelaria baicalensis, Zea mays
Species of your interest not listed? Contact us

Not reactive in

Chlamydomonas reinhardtii

Application examples

Application examples 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 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 chemiluminescent detection reagent of extreme low femtogram range and visualized by exposing to a film (Agfa Cronex 5) for 5 min.

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

Additional information

Total IgY concentration is 30.8 mg/ml

This 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.

Related products

Collection of antibodies for plant developmental biology

Plant protein extraction buffer

Secondary antibodies

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

Product citations

Selected references Leviczky et al. (2019). E2FA and E2FB transcription factors coordinate cell proliferation with seed maturation. Development. 2019 Nov 26;146(22). pii: dev179333. doi: 10.1242/dev.179333.
Horvath et al. (2017). Arabidopsis RETINOBLASTOMA RELATED directly regulates DNA damage responses through functions beyond cell cycle control. EMBO J. 2017 May 2;36(9):1261-1278. doi: 10.15252/embj.201694561. Epub 2017 Mar 20.
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.