Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply Rabbit monoclonal [EPR17512] to Rb Suitable for: Flow Cyt (Intra), WB, IP, ICC/IF, IHC-P Knockout validated Reacts with: Mouse, Human, African green monkey

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

WB: Jurkat, Hek294, K562, WEHI-3, COS-1, MCF7 and F9 whole cell lysates; Mouse brain and lung lysates; Human fetal brain lysate.IHC-P: Human lung, Human breast cancer, Mouse lung and Mouse cerebral cortex tissues.ICC/IF: MCF7 cells.IP: MCF7 whole cell lysate.
常规说明

This product is a recombinant monoclonal antibody, which offers several advantages including:- High batch-to-batch consistency and reproducibility- Improved sensitivity and specificity- Long-term security of supply- Animal-free productionFor more information see here.

Our RabMAb technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb patents.

We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated \'PUR\' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

存放说明 Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
存储溶液 Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
The Abpromise guarantee

Abpromise™承诺保证使用ab181616于以下的经测试应用

\"应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Key regulator of entry into cell division that acts as a tumor suppressor. Promotes G0-G1 transition when phosphorylated by CDK3/cyclin-C. Acts as a transcription repressor of E2F1 target genes. The underphosphorylated, active form of RB1 interacts with E2F1 and represses its transcription activity, leading to cell cycle arrest. Directly involved in heterochromatin formation by maintaining overall chromatin structure and, in particular, that of constitutive heterochromatin by stabilizing histone methylation. Recruits and targets histone methyltransferases SUV39H1, KMT5B and KMT5C, leading to epigenetic transcriptional repression. Controls histone H4 \'Lys-20\' trimethylation. Inhibits the intrinsic kinase activity of TAF1. Mediates transcriptional repression by SMARCA4/BRG1 by recruiting a histone deacetylase (HDAC) complex to the c-FOS promoter. In resting neurons, transcription of the c-FOS promoter is inhibited by BRG1-dependent recruitment of a phospho-RB1-HDAC1 repressor complex. Upon calcium influx, RB1 is dephosphorylated by calcineurin, which leads to release of the repressor complex (By similarity). In case of viral infections, interactions with SV40 large T antigen, HPV E7 protein or adenovirus E1A protein induce the disassembly of RB1-E2F1 complex thereby disrupting RB1\'s activity.
The Pocket domain binds to the threonine-phosphorylated domain C, thereby preventing interaction with heterodimeric E2F/DP transcription factor complexes.
翻译后修饰 Phosphorylated by CDK6 and CDK4, and subsequently by CDK2 at Ser-567 in G1, thereby releasing E2F1 which is then able to activate cell growth. Dephosphorylated at the late M phase. SV40 large T antigen, HPV E7 and adenovirus E1A bind to the underphosphorylated, active form of pRb. Phosphorylation at Thr-821 and Thr-826 promotes interaction between the C-terminal domain C and the Pocket domain, and thereby inhibits interactions with heterodimeric E2F/DP transcription factor complexes. Dephosphorylated at Ser-795 by calcineruin upon calcium stimulation. CDK3/cyclin-C-mediated phosphorylation at Ser-807 and Ser-811 is required for G0-G1 transition. Phosphorylated by CDK1 and CDK2 upon TGFB1-mediated apoptosis.
N-terminus is methylated by METTL11A/NTM1 (By similarity). Monomethylation at Lys-810 by SMYD2 enhances phosphorylation at Ser-807 and Ser-811, and promotes cell cycle progression. Monomethylation at Lys-860 by SMYD2 promotes interaction with L3MBTL1.
Acetylation at Lys-873 and Lys-874 regulates subcellular localization, at least during keratinocytes differentiation.
Exon 17 tumor GOS561 substitution mutation causes premature stop antibody GOS563 exon 17 substitution mutation causes premature stop antibody OSRC antibody
Prepro retinoblastoma associated protein antibody Protein phosphatase 1 regulatory subunit 130 antibody Rb antibody RB transcriptional corepressor 1 antibody RB_HUMAN antibody RB1 antibody RB1 gene antibody Retinoblastoma 1 antibody Retinoblastoma suspectibility protein antibody Retinoblastoma-associated protein antibody see all

Immunohistochemical analysis of paraffin-embedded Human lung tissue labeling Rb with ab181616 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on Human lung is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Lane 1: Wild-type HAP1 whole cell lysate (20 g)
Lane 2: Rb knockout HAP1 whole cell lysate (20 g)
Lane 3: HeLa whole cell lysate (20 g)
Lane 4: HEK293 whole cell lysate (20 g)

Lanes 1 - 4: Merged signal (red and green). Green - ab181616 observed at 120 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab181616 was shown to recognize Rb in wild-type HAP1 cells as signal was lost at the expected MW in Rb knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and RB1 knockout samples were subjected to SDS-PAGE. Ab181616 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4 C at 1/2000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H L (IRDye 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H L (IRDye 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MCF7 (Human breast adenocarcinoma cell line) cells labeling Rb with ab181616 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing nuclear staining on MCF7 cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor 594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:
-ve control 1: ab181616 at 1/500 dilution followed by ab150120 (AlexaFluor 594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor 488 Goat Anti-Rabbit IgG H L) at 1/500 dilution.

ab181616 staining Rbin the human cell line MCF-7(human breast carcinoma) by flow cytometry. Cells were fixed with 4% paraformaldehyde, permeabilized with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/140. A goat anti rabbit IgG (Alexa Fluor 488) at a dilution of 1/2000 was used as the secondary antibody.

Isoytype control: Rabbit monoclonal IgG (Black)

Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

All lanes : Anti-Rb antibody [EPR17512] (ab181616) at 1/1000 dilution

Lane 1 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates prepared in RIPA lysis method with 5% NFDM/TBST
Lane 2 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates prepared in 1%SDS Hot lysis method with 5% NFDM/TBST
Lane 3 : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysates prepared in RIPA lysis method with 5% NFDM/TBST
Lane 4 : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysates prepared in 1%SDS Hot lysis method with 5% NFDM/TBST

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

Predicted band size: 105 kDa

Exposure time:

Lane 1 and 2: 100 seconds
Lane 3 and 4: 10 seconds

We recommend to use 1%SDS Hot lysis method to get clear band.
We are unsure how to define the extra bands.

Rb was immunoprecipitated from 1mg of MCF7 (Human breast adenocarcinoma cell line) whole cell lysate with ab181616 at 1/80 dilution. Western blot was performed from the immunoprecipitate using ab181616 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

Lane 1: MCF7 whole cell lysate 10 g (Input). Lane 2: ab181616 IP in MCF7 whole cell lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab181616 in MCF7 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.

All lanes : Anti-Rb antibody [EPR17512] (ab181616) at 1/20000 dilution

Lane 1 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate
Lane 2 : K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysate
Lane 3 : WEHI-3 (Mouse leukemia) whole cell lysate
Lane 4 : COS-1 (African green monkey kidney fibroblast-like cell line) whole cell lysate
Lane 5 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 105 kDa
Observed band size: 105 kDa


Exposure time: 5 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.

The lysates were all prepared using 1%SDS Hot lysis method.

All lanes : Anti-Rb antibody [EPR17512] (ab181616) at 1/10000 dilution

Lane 1 : F9 (Mouse embyro testicular cancer cell line) whole cell lysate
Lane 2 : Mouse brain lysate
Lane 3 : Mouse lung lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 105 kDa
Observed band size: 105 kDa


Exposure time: 1 minute

Blocking/Dilution buffer: 5% NFDM/TBST.

The lysates were all prepared using 1%SDS Hot lysis method.

Anti-Rb antibody [EPR17512] (ab181616) at 1/2000 dilution + Human fetal brain lysate at 10 µg

Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 105 kDa
Observed band size: 105 kDa


Exposure time: 1 minute

Blocking/Dilution buffer: 5% NFDM/TBST.

The lysates were all prepared using 1%SDS Hot lysis method.

Immunohistochemical analysis of paraffin-embedded Human breast cancertissue labeling Rb with ab181616 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on cancer cells of Human breast canceris observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling Rb with ab181616 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on mouse lung is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Immunohistochemical analysis of paraffin-embedded Mouse cerebral cortextissue labeling Rb with ab181616 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on neuron of mouse cerebral cortexis observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

To download a Certificate of Compliance, please enter your Lot number below:

Lot number

发表研究结果有使用 ab181616？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab181616 被引用在 39 文献中.

Wang L et al. A genetically defined disease model reveals that urothelial cells can initiate divergent bladder cancer phenotypes. Proc Natl Acad Sci U S A 117:563-572 (2020).PubMed: 31871155 Wang Z et al. Nuclear receptor HNF4a performs a tumor suppressor function in prostate cancer via its induction of p21-driven cellular senescence. Oncogene 39:1572-1589 (2020).PubMed: 31695151 Li S et al. Pan-cancer analysis reveals synergistic effects of CDK4/6i and PARPi combination treatment in RB-proficient and RB-deficient breast cancer cells. Cell Death Dis 11:219 (2020).PubMed: 32249776 Ma Y et al. Fli-1 Activation through Targeted Promoter Activity Regulation Using a Novel 3\', 5\'-diprenylated Chalcone Inhibits Growth and Metastasis of Prostate Cancer Cells. Int J Mol Sci 21:N/A (2020).PubMed: 32210104 Ji W et al. c-myc regulates the sensitivity of breast cancer cells to palbociclib via c-myc/miR-29b-3p/CDK6 axis. Cell Death Dis 11:760 (2020).PubMed: 32934206 View all Publications for this product

There are currently no Customer reviews or Questions for ab181616.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES
For licensing inquiries, please contact partnerships@abcam.com