RBR1 | Retinoblastoma related protein

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

Limited stock

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 Purified, total IgY (chicken egg yolk immunoglobulin) in PBS pH 8. Contains 0.02 % sodium azide.

Format Liquid

Quantity 100 µl

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

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

Reactant: Arabidopsis thaliana (Thale cress)

Application: Western Blotting

Pudmed ID: 31666236

Journal: Development

Figure Number: 1C

Published Date: 2019-11-26

First Author: Leviczky, T., Molnár, E., et al.

Impact Factor: 5.272

Open Publication

The expression profiles of E2FA and E2FB are distinct in the developing siliques, but overlap in the proliferation phase. (A) qRT-PCR analyses of the G2- and M-phase-specific CDKB1;1 and the seed maturation LEC2 and WRI1 genes in the developing siliques of the wild-type (WT) at four silique developmental stages (S1-S4, pictured in Fig. S1). (B) The transcript levels of the three E2Fs, namely E2FA, E2FB and E2FC, and the single RBR genes were also analysed in these silique samples by qRT-PCR. Values represent fold-changes normalised to the value of the S1 silique stage (set arbitrarily at 1). Data are mean±s.d., n=3 biological repeats. *P?0.05, **P?0.01, ***P?0.001, ****P?0.0001 (two-tailed, paired t-test between consecutive silique stages). ns, non-significant. (C) To follow the accumulation levels of RBR, E2FA, E2FB and DPB proteins in developing siliques (S1-S4) specific antibodies were used in immunoblot assays as indicated. The Ponceau-stained proteins were used as loading control. Arrowheads indicate the corresponding E2FA and DPB proteins; arrow marks a slower migrating form of DPB in S4 silique stage. Molecular weights of the specific proteins are shown on the left.

Additional information

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.