Phospho-Retinoblastoma (Ser780) Antibody | Affinity Biosciences LTD|亲科生物官网

产品:	磷酸化 Retinoblastoma (Ser780) 抗体
货号:	AF3103
描述:	Rabbit polyclonal antibody to Phospho-Retinoblastoma (Ser780)
应用:	WB IHC IF/ICC
反应:	Human, Mouse, Rat
预测:	Pig, Bovine, Horse, Sheep, Rabbit, Dog, Chicken, Xenopus
分子量:	106kDa; 106kD(Calculated).
蛋白号:	P06400
RRID:	AB_2834540

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阻断肽(封闭肽)是特异性结合目标抗体和阻断抗体结合的多肽。这些多肽包含抗体识别的抗原表位。与阻断肽结合的抗体不再与靶蛋白上的表位结合。例如，在免疫印迹(WB)和免疫组化(IHC)中，通过比较被阻断抗体和未阻断抗体的染色，可以看到哪种染色是特异性的。

规格	价格	库存
50ul	RMB¥ 1300	现货
100ul	RMB¥ 2400	现货
200ul	RMB¥ 3200	现货

货期: 当天发货

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实验方案

产品描述

来源:

Rabbit

应用:

WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: 适用于变性蛋白样本的免疫印迹检测. IHC: 适用于组织样本的石蜡(IHC-p)或冰冻(IHC-f)切片样本的免疫组化/荧光检测. IF/ICC: 适用于细胞样本的荧光检测. ELISA(peptide): 适用于抗原肽的ELISA检测.

反应:

Human,Mouse,Rat

预测:

Pig(100%), Bovine(100%), Horse(91%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(91%), Xenopus(90%)

克隆:

Polyclonal

特异性:

Phospho-Retinoblastoma (Ser780) Antibody detects endogenous levels of Retinoblastoma only when phosphorylated at Serine 780.

RRID:

AB_2834540
引用格式: Affinity Biosciences Cat# AF3103, RRID:AB_2834540.

偶联:

Unconjugated.

纯化:

The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.

保存:

Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.

别名:

展开/折叠

Exon 17 tumor GOS561 substitution mutation causes premature stop; GOS563 exon 17 substitution mutation causes premature stop; OSRC; Osteosarcoma; p105-Rb; P105RB; PP105; pp110; PPP1R130; pRb; Prepro retinoblastoma associated protein; Protein phosphatase 1 regulatory subunit 130; Rb; RB transcriptional corepressor 1; RB_HUMAN; RB1; RB1 gene; Retinoblastoma 1; Retinoblastoma suspectibility protein; Retinoblastoma-associated protein;

抗原和靶标

免疫原:

Uniprot:

P06400(RB_HUMAN) >>Visit HPA database.

基因/基因ID:

RB1(5925)

表达:

P06400 RB_HUMAN:

Expressed in the retina. Expressed in foreskin keratinocytes (at protein level) (PubMed:20940255).

描述:

Retinoblastoma (RB) is an embryonic malignant neoplasm of retinal origin. It almost always presents in early childhood and is often bilateral. Spontaneous regression ('cure') occurs in some cases.

序列:

P06400 RB_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

MPPKTPRKTAATAAAAAAEPPAPPPPPPPEEDPEQDSGPEDLPLVRLEFEETEEPDFTALCQKLKIPDHVRERAWLTWEKVSSVDGVLGGYIQKKKELWGICIFIAAVDLDEMSFTFTELQKNIEISVHKFFNLLKEIDTSTKVDNAMSRLLKKYDVLFALFSKLERTCELIYLTQPSSSISTEINSALVLKVSWITFLLAKGEVLQMEDDLVISFQLMLCVLDYFIKLSPPMLLKEPYKTAVIPINGSPRTPRRGQNRSARIAKQLENDTRIIEVLCKEHECNIDEVKNVYFKNFIPFMNSLGLVTSNGLPEVENLSKRYEEIYLKNKDLDARLFLDHDKTLQTDSIDSFETQRTPRKSNLDEEVNVIPPHTPVRTVMNTIQQLMMILNSASDQPSENLISYFNNCTVNPKESILKRVKDIGYIFKEKFAKAVGQGCVEIGSQRYKLGVRLYYRVMESMLKSEEERLSIQNFSKLLNDNIFHMSLLACALEVVMATYSRSTSQNLDSGTDLSFPWILNVLNLKAFDFYKVIESFIKAEGNLTREMIKHLERCEHRIMESLAWLSDSPLFDLIKQSKDREGPTDHLESACPLNLPLQNNHTAADMYLSPVRSPKKKGSTTRVNSTANAETQATSAFQTQKPLKSTSLSLFYKKVYRLAYLRLNTLCERLLSEHPELEHIIWTLFQHTLQNEYELMRDRHLDQIMMCSMYGICKVKNIDLKFKIIVTAYKDLPHAVQETFKRVLIKEEEYDSIIVFYNSVFMQRLKTNILQYASTRPPTLSPIPHIPRSPYKFPSSPLRIPGGNIYISPLKSPYKISEGLPTPTKMTPRSRILVSIGESFGTSEKFQKINQMVCNSDRVLKRSAEGSNPPKPLKKLRFDIEGSDEADGSKHLPGESKFQQKLAEMTSTRTRMQKQKMNDSMDTSNKEEK

种属预测

种属预测:

score>80的预测可信度较高，可尝试用于WB检测。*预测模型主要基于免疫原序列比对，结果仅作参考，不作为质保凭据。

Species

Results

Score

Pig

100

Bovine

100

Sheep

100

Dog

100

Rabbit

100

Horse

Chicken

Xenopus

Zebrafish

Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

翻译修饰 - P06400 作为底物

P06400

Site	PTM Type	Enzyme	Source
	Phosphorylation		Uniprot
T5	Phosphorylation	P11802 (CDK4)	Uniprot
S37	Phosphorylation		Uniprot
K63	Ubiquitination		Uniprot
K65	Ubiquitination		Uniprot
K94	Ubiquitination		Uniprot
K136	Ubiquitination		Uniprot
T140	Phosphorylation		Uniprot
K143	Ubiquitination		Uniprot
S149	Phosphorylation		Uniprot
S163	Phosphorylation		Uniprot
S230	Phosphorylation	P24941 (CDK2)	Uniprot
Y239	Phosphorylation		Uniprot
T241	Phosphorylation		Uniprot
S249	Phosphorylation	P24941 (CDK2) , P11802 (CDK4) , P05771 (PRKCB) , P06493 (CDK1)	Uniprot
T252	Phosphorylation	P06493 (CDK1) , P05771 (PRKCB) , P24941 (CDK2) , P11802 (CDK4)	Uniprot
K265	Ubiquitination		Uniprot
K279	Ubiquitination		Uniprot
K289	Ubiquitination		Uniprot
K319	Ubiquitination		Uniprot
Y321	Phosphorylation		Uniprot
Y325	Phosphorylation		Uniprot
K327	Ubiquitination		Uniprot
K341	Ubiquitination		Uniprot
T345	Phosphorylation		Uniprot
S347	Phosphorylation		Uniprot
S350	Phosphorylation		Uniprot
T353	Phosphorylation		Uniprot
T356	Phosphorylation	P24941 (CDK2) , P11802 (CDK4)	Uniprot
K359	Ubiquitination		Uniprot
S360	Phosphorylation		Uniprot
T373	Phosphorylation	P06493 (CDK1) , P24941 (CDK2) , P11802 (CDK4)	Uniprot
K420	Ubiquitination		Uniprot
K427	Acetylation		Uniprot
K427	Ubiquitination		Uniprot
K432	Ubiquitination		Uniprot
Y454	Phosphorylation		Uniprot
K537	Ubiquitination		Uniprot
K548	Acetylation		Uniprot
S567	Phosphorylation	Q16539 (MAPK14) , P24941 (CDK2)	Uniprot
K574	Ubiquitination		Uniprot
T583	Phosphorylation		Uniprot
S588	Phosphorylation		Uniprot
T601	Phosphorylation		Uniprot
Y606	Phosphorylation		Uniprot
S608	Phosphorylation	P24941 (CDK2) , P11802 (CDK4)	Uniprot
S612	Phosphorylation	O96017 (CHEK2) , O14757 (CHEK1) , Q00534 (CDK6) , P11802 (CDK4) , P24941 (CDK2)	Uniprot
S618	Phosphorylation		Uniprot
S624	Phosphorylation		Uniprot
T625	Phosphorylation		Uniprot
K640	Ubiquitination		Uniprot
K720	Sumoylation		Uniprot
Y771	Phosphorylation		Uniprot
S773	Phosphorylation		Uniprot
T774	Phosphorylation		Uniprot
R775	Methylation		Uniprot
T778	Phosphorylation		Uniprot
S780	Phosphorylation	P11802 (CDK4) , P30279 (CCND2) , P17612 (PRKACA) , Q00534 (CDK6) , Q96GD4 (AURKB) , P05771 (PRKCB)	Uniprot
R787	Methylation		Uniprot
S788	Phosphorylation	Q00534 (CDK6) , P11802 (CDK4)	Uniprot
Y790	Phosphorylation		Uniprot
K791	Ubiquitination		Uniprot
S794	Phosphorylation		Uniprot
S795	Phosphorylation	P05771 (PRKCB) , P11802 (CDK4) , P50750 (CDK9) , Q00534 (CDK6) , Q00535 (CDK5)	Uniprot
R798	Methylation		Uniprot
Y805	Phosphorylation	P00519 (ABL1)	Uniprot
S807	Phosphorylation	P24941 (CDK2) , P50750 (CDK9) , Q00534 (CDK6) , P11802 (CDK4) , Q00526 (CDK3) , P06493 (CDK1) , Q00535 (CDK5)	Uniprot
K810	Methylation		Uniprot
K810	Ubiquitination		Uniprot
S811	Phosphorylation	Q00534 (CDK6) , P06493 (CDK1) , P24941 (CDK2) , P11802 (CDK4) , Q00526 (CDK3) , P50750 (CDK9) , Q00535 (CDK5) , Q13131 (PRKAA1)	Uniprot
Y813	Phosphorylation		Uniprot
K814	Ubiquitination		Uniprot
S816	Phosphorylation		Uniprot
T821	Phosphorylation	P24941 (CDK2) , Q00534 (CDK6) , P05771 (PRKCB)	Uniprot
T823	Phosphorylation		Uniprot
K824	Ubiquitination		Uniprot
T826	Phosphorylation	Q00534 (CDK6) , P11802 (CDK4) , P24941 (CDK2)	Uniprot
S829	Phosphorylation		Uniprot
S834	Phosphorylation		Uniprot
S838	Phosphorylation		Uniprot
T841	Phosphorylation		Uniprot
S842	Phosphorylation		Uniprot
K844	Ubiquitination		Uniprot
K847	Ubiquitination		Uniprot
S855	Phosphorylation		Uniprot
R857	Methylation		Uniprot
K860	Methylation		Uniprot
K860	Ubiquitination		Uniprot
S862	Phosphorylation		Uniprot
S866	Phosphorylation		Uniprot
K870	Ubiquitination		Uniprot
K873	Acetylation		Uniprot
K874	Acetylation		Uniprot
S882	Phosphorylation		Uniprot
S895	Phosphorylation		Uniprot
K896	Acetylation		Uniprot
K896	Ubiquitination		Uniprot
K900	Ubiquitination		Uniprot
T905	Phosphorylation		Uniprot
S919	Phosphorylation		Uniprot
K925	Acetylation		Uniprot

研究背景

P06400_RB_HUMAN

功能:

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).

(Microbial infection) 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.

翻译修饰:

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 (By similarity).

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.

Acetylated during keratinocyte differentiation. Acetylation at Lys-873 and Lys-874 regulates subcellular localization. Can be deacetylated by SIRT1.

细胞定位:

Nucleus.
Note: During keratinocyte differentiation, acetylation by KAT2B/PCAF is required for nuclear localization.

组织特异性:

Expressed in the retina. Expressed in foreskin keratinocytes (at protein level).

亚基结构:

Interacts with ATAD5. Interacts with PRMT2, CDK1 and CDK2 (By similarity). The hypophosphorylated form interacts with and sequesters the E2F1 transcription factor. Interacts with heterodimeric E2F/DP transcription factor complexes containing TFDP1 and either E2F1, E2F3, E2F4 or E2F5, or TFDP2 and E2F4. The unphosphorylated form interacts with EID1, ARID3B, KDM5A, SUV39H1, MJD2A/JHDM3A and THOC1. Interacts with the N-terminal domain of TAF1. Interacts with SNW1, AATF, DNMT1, LIN9, LMNA, KMT5B, KMT5C, PELP1, UHRF2 and TMPO-alpha. May interact with NDC80. Interacts with GRIP1 and UBR4. Interacts with ARID4A and KDM5B. Interacts with E4F1 and LIMD1. Interacts with SMARCA4/BRG1 AND HDAC1 (By similarity). Interacts with PSMA3 and USP4. Interacts (when methylated at Lys-860) with L3MBTL1. Interacts with CHEK2; phosphorylates RB1. Interacts with CEBPA. P-TEFB complex interacts with RB1; promotes phosphorylation of RB1. Interacts with RBBP9; the interaction disrupts RB1 binding to E2F1 (By similarity). Interacts with KAT2B/PCAF and EP300/P300 (By similarity).

(Microbial infection) Interacts with adenovirus E1A protein.

(Microbial infection) Interacts with HPV E7 protein.

(Microbial infection) Interacts with SV40 large T antigen.

(Microbial infection) Interacts with human cytomegalovirus/HHV-5 protein UL123.

(Microbial infection) Interacts with molluscum contagiosum virus protein MC007.

蛋白家族:

The Pocket domain binds to the threonine-phosphorylated domain C, thereby preventing interaction with heterodimeric E2F/DP transcription factor complexes.

Belongs to the retinoblastoma protein (RB) family.

研究领域

· Cellular Processes > Cell growth and death > Cell cycle. (View pathway)

· Cellular Processes > Cell growth and death > Cellular senescence. (View pathway)

· Human Diseases > Drug resistance: Antineoplastic > Endocrine resistance.

· Human Diseases > Infectious diseases: Viral > Hepatitis B.

· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.

· Human Diseases > Infectious diseases: Viral > HTLV-I infection.

· Human Diseases > Infectious diseases: Viral > Epstein-Barr virus infection.

· Human Diseases > Cancers: Overview > Pathways in cancer. (View pathway)

· Human Diseases > Cancers: Overview > Viral carcinogenesis.

· Human Diseases > Cancers: Specific types > Pancreatic cancer. (View pathway)

· Human Diseases > Cancers: Specific types > Glioma. (View pathway)

· Human Diseases > Cancers: Specific types > Prostate cancer. (View pathway)

· Human Diseases > Cancers: Specific types > Melanoma. (View pathway)

· Human Diseases > Cancers: Specific types > Bladder cancer. (View pathway)

· Human Diseases > Cancers: Specific types > Chronic myeloid leukemia. (View pathway)

· Human Diseases > Cancers: Specific types > Small cell lung cancer. (View pathway)

· Human Diseases > Cancers: Specific types > Non-small cell lung cancer. (View pathway)

· Human Diseases > Cancers: Specific types > Breast cancer. (View pathway)

· Human Diseases > Cancers: Specific types > Hepatocellular carcinoma. (View pathway)

· Human Diseases > Cancers: Specific types > Gastric cancer. (View pathway)

文献引用

1). Blockage of EGFR/AKT and mevalonate pathways synergize the antitumor effect of temozolomide by reprogramming energy metabolism in glioblastoma. Cancer communications (London, England), 2023 (PubMed: 37920878) [IF=16.2]

Application: WB Species: Human Sample: TBD0220, U-87 MG, and U-87 MG-EGFR-vIII cells

FIGURE 3 EGFR/AKT pathway regulates mitochondrial respiration and proliferation in GBM cells. (A-B) TBD0220 (A), and U-87 MG-EGFR-vIII cells (B) were treated with DMSO or 5 μmol/L MK-2206 for 24 h. The mitochondrial functions were monitored by Seahorse XF Cell Mito Stress test. The OCR, basal respiration, proton leak, and ATP production rates were measured as illustrated. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001 (independent-sample Student's t-test for TBD0220; one-way ANOVA for U-87 MG). (C) ATP levels in TBD0220, U-87 MG, and U-87 MG-EGFR-vIII cells were analyzed after 24 h of treatments with DMSO or 5 μmol/L of MK-2206. ∗∗P < 0.01, ∗∗∗P < 0.001 (independent-sample Student's t-test for TBD0220; one-way ANOVA for U-87 MG). (D) The cell growth assay for TBD0220, U-87 MG, and U-87 MG-EGFR-vIII lines treated with DMSO, 5 μmol/L MK-2206, or 5 μmol/L MK-2206 plus 50 μmol/L ATP were performed. ∗∗∗P < 0.001 (two-way ANOVA). (E) The colony formation assay of TBD0220, U-87 MG, and U-87 MG-EGFR-vIII lines treated with DMSO or 1 μmol/L MK-2206. (F) Cell cycle distributions were analyzed by flow cytometry in TBD0220, U-87 MG, and U-87 MG-EGFR-vIII cells treated with DMSO or 5 μmol/L MK-2206. (G) Western blotting to show changes in expressions of CDK2, CDK4, CDK6, Cyclin D, RB, p-RB, and GAPDH in TBD0220, U-87 MG, and U-87 MG-EGFR-vIII cells treated with DMSO or MK-2206. Abbreviations: EGFR, epidermal growth factor receptor; AKT, AKT serine/threonine kinase 1; GBM, glioblastoma; ANOVA, analysis of variance; DMSO, dimethyl sulfoxide; CDK2, cyclin-dependent kinase 2; CDK4, cyclin-dependent kinase 4; CDK6, cyclin-dependent kinase 6; OCR, oxygen consumption rate.

2). TCA-phospholipid-glycolysis targeted triple therapy effectively suppresses ATP production and tumor growth in glioblastoma. Theranostics, 2023 (PubMed: 36276638) [IF=12.4]

Application: WB Species: Human Sample: TBD0220 and U87MG cells

Figure 3 Reduction in ATP production hinders cell proliferation and contributes to G1/S arrest in GBM. (A-B) The relative viability of TBD0220 (A) and U87MG (B) cells was measured using a CCK-8 kit (n = 3). (C-D) Colony formation assay to detect GBM cell growth (D), and the quantification of colony numbers (C) (n = 3). (E) Cell cycle analysis using flow cytometry after incubation with different treatments like EPIC (20 µM), AA (25 µM), or EPIC (20 µM) + AA (25 µM) for 48 h, and the results are plotted as a histogram (n = 3) (F) Representative western blotting showing the expression of p21 and Rb and their downstream targets. The results were normalized to Tubulin with the control group as 1. Protein expression was quantified by ImageJ. (G) Representative confocal images of CDK6 after the treatment with AA (25 µM), EPIC (20 µM), or EPIC (20 µM) + AA (25 µM) for 48 h (n = 6). (H) The quantitative analysis of fluorescence images of CDK6 (n = 6). All data are shown as the mean values ± SD, and p values are based on one-way or two-way ANOVA. ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05. Scale bar = 20 μm.

3). Combination LSD1 and HOTAIR-EZH2 inhibition disrupts cell cycle processes and induces apoptosis in glioblastoma cells. PHARMACOLOGICAL RESEARCH, 2021 (PubMed: 34246782) [IF=9.3]

4). Metformin as a senostatic drug enhances the anticancer efficacy of CDK4/6 inhibitor in head and neck squamous cell carcinoma. Cell Death & Disease, 2020 (PubMed: 33116117) [IF=9.0]

Application: WB Species: Human Sample:

Fig. 2 CDK4/6 inhibitor combined with metformin promoted cell cycle arrest. A, B Flow cytometry results showed that the combination of the CDK4/6 inhibitor LY2835219 (HSC3: 0.1 μM; HSC6: 0.3 μM; Cal27: 1.25 μM) with metformin (HSC3: 10 mM; HSC6: 1 mM; Cal27: 10 mM) induced cell cycle arrest in the G0/G1 phase. C Western blot results showed that LY2835219 upregulated the expression of p16 and p21 and downregulated the expression of pRb. D Western blot results showed that metformin upregulated the expression of p16 and p21 and downregulated the expression of pRb. E Compared with LY2835219 monotherapy, the combination treatment significantly upregulated p21 expression and suppressed p-Rb expression. F Immunohistochemistry results (400×) showed that LY2835219 and metformin synergistically upregulated p21 expression and downregulated pRb expression in HSC6 xenograft tumors. Bar: 100 μm. *P

Application: IHC Species: Human Sample:

5). Combination of N, N′-dicyclohexyl-N-arachidonic acylurea and tacrolimus prolongs cardiac allograft survival in mice. Immunology and Cell Biology, 2020 (PubMed: 32162358) [IF=4.0]

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Phospho-Retinoblastoma (Ser780) Antibody - #AF3103

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