Phospho-Rad17 (Ser656) Antibody (YA2640)

Cat. No.: HY-P82895: Data Sheet COA
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Phospho-Rad17 (Ser656) Antibody (YA2640) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to Phospho-Rad17 (Ser656).

For research use only. We do not sell to patients.

Size	Stock	Quantity
10 μL	Get quote 2 - 3 Weeks 1 - 2 Weeks 3 - 4 Weeks 2 - 3 weeks	Estimated Time of Arrival: December 31
50 μL	Get quote 2 - 3 Weeks 1 - 2 Weeks 3 - 4 Weeks 2 - 3 weeks	Estimated Time of Arrival: December 31
100 μL	Get quote 2 - 3 Weeks 1 - 2 Weeks 3 - 4 Weeks 2 - 3 weeks	Estimated Time of Arrival: December 31
250 μL	Get quote

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Phospho-Rad17 (Ser656) Antibody (YA2640) Featured Recommendations:

WB: Western Blot;
IHC-P: Immunohistochemistry-Paraffin;
IHC-F: Immunohistochemistry-Frozen;
ICC/IF: Immunocytochemistry/Immunofluorescence;
IF-Tissue: Immunofluorescence-Tissue;
mIHC: Multiplex Immunohistochemical;
IP: Immunoprecipitation;
ChIP: Chromatin Immunoprecipitation;
FC: Flow Cytometry;
ELISA: Enzyme Linked Immunosorbent Assay

Product Detail
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Background
Documentation

Description

Phospho-Rad17 (Ser656) Antibody (YA2640) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to Phospho-Rad17 (Ser656).

Host

Rabbit

Clonality

Recombinant,Monoclonal

Molecular Weight

Predicted band size: 77 kDa;

Observed band size: 80 kDa

Note: Due to possible protein modifications or aggregation, the molecular weight should be confirmed by actual measurement, and the predicted value is for reference only.

Species Reactivity

Human

SwissProt ID

O75943

Gene ID

5884 [NCBI]

Immunogen

A synthetic phosphopeptide corresponding to residues surrounding Ser656 of human Rad17

Application &
Dilution Ratio

Application	Dilution Ratio
WB WB: Western Blot	1:500-1:1000
IHC-P IHC-P: Immunohistochemistry-Paraffin	1:50-1:100
IHC-F IHC-F: Immunohistochemistry-Frozen	1:50-1:100
ICC/IF ICC/IF: Immunocytochemistry/Immunofluorescence	1:50-1:200

Sensitivity	Endogenous	Purity	Affinity Purified
Conjugation	Non-conjugated	Modification	Phosphorylated
Isotype	IgG

Appearance

Liquid

Formulation

Supplied in 50mM Tris-Glycine(pH 7.4), 0.15M NaCl, 40% Glycerol, 0.01% Sodium azide and 0.05% BSA

Storage & Stability

Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

Shipping

Shipping with blue ice.

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ALL WB IHC-P mIHC

Western blot analysis was performed on extracts from Hela (lane 1, 15 μg), Hela+CA (lane 2, 15 μg), HepG2 (lane 3, 15 μg), Jurkat (lane 4, 15 μg), and Jurkat+Etoposide (lane 5, 15 μg) using Phospho-Rad17 (Ser656) Rabbit mAb.Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST at 4°C overnight.The primary antibody (1:1000 dilution) and the loading control antibody (beta-Tubulin, HY-P80955, 1:5000 dilution) was incubated in 5% non-fat milk in TBST for 1 hour at 37°C.Goat Anti-Rabbit IgG-HRP Secondary Antibody (1:20000 dilution) was then applied for 40 minutes at 37°C.
Immunohistochemical analysis of paraffin-embedded human Testis‌ tissue using Phospho-Rad17 (Ser656) antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P82895, 1:50 dilution) at room temperature for 60 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.
Immunohistochemical analysis of paraffin-embedded human Testis tissue using Phospho-Rad17 (Ser656) antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P82895, 1:50 dilution) at room temperature for 60 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.
Immunohistochemical analysis of paraffin-embedded human Breast Cancer‌ tissue using Phospho-Rad17 (Ser656) antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P82895, 1:50 dilution) at room temperature for 120 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.
Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma‌ tissue using Phospho-Rad17 (Ser656) antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P82895, 1:50 dilution) at room temperature for 120 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.
Immunohistochemical analysis of paraffin-embedded human Cervical cancer tissue using Phospho-Rad17 (Ser656) antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P82895, 1:50 dilution) at room temperature for 120 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.
Immunohistochemical analysis of paraffin-embedded human Testis‌ tissue using Phospho-Rad17 (Ser656) antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P82895, 1:50 dilution) at room temperature for 120 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.
Tyramide signaling amplification based immunofluorescence was performed on paraffin-embedded human Breast Cancer ‌ tissue using Phospho-Rad17 (Ser656) antibody. Antigen retrieval was performed in EDTA buffer pH 9.0 (95 °C, 20 min) followed by cooling to RT. Then incubated with primary antibody (HY-P82895, 1:50 dilution) at room temperature for 60 minutes and HRP conjugated secondary antibody for 10 minutes. Fluorescence was then developed with Vari Fluor 532 TSA (200×)（HY-D1832）. The tissues were counterstained with DAPI and mounted with Anti-fade mounting medium.
Tyramide signaling amplification based immunofluorescence was performed on paraffin-embedded human Breast Cancer ‌ tissue using Phospho-Rad17 (Ser656) antibody. Antigen retrieval was performed in EDTA buffer pH 9.0 (95 °C, 20 min) followed by cooling to RT. Then incubated with primary antibody (HY-P82895, 1:50 dilution) at room temperature for 60 minutes and HRP conjugated secondary antibody for 10 minutes. Fluorescence was then developed with Vari Fluor 532 TSA (200×)（HY-D1832）. The tissues were counterstained with DAPI and mounted with Anti-fade mounting medium.
Tyramide signaling amplification based immunofluorescence was performed on paraffin-embedded human Breast Cancer ‌ tissue using Phospho-Rad17 (Ser656) antibody. Antigen retrieval was performed in EDTA buffer pH 9.0 (95 °C, 20 min) followed by cooling to RT. Then incubated with primary antibody (HY-P82895, 1:50 dilution) at room temperature for 60 minutes and HRP conjugated secondary antibody for 10 minutes. Fluorescence was then developed with Vari Fluor 532 TSA (200×)（HY-D1832）. The tissues were counterstained with DAPI and mounted with Anti-fade mounting medium.
Tyramide signaling amplification based immunofluorescence was performed on paraffin-embedded human Testis tissue using Phospho-Rad17 (Ser656) antibody. Antigen retrieval was performed in EDTA buffer pH 9.0 (95 °C, 20 min) followed by cooling to RT. Then incubated with primary antibody (HY-P82895, 1:50 dilution) at room temperature for 60 minutes and HRP conjugated secondary antibody for 10 minutes. Fluorescence was then developed with Vari Fluor 532 TSA (200×)（HY-D1832）. The tissues were counterstained with DAPI and mounted with Anti-fade mounting medium.
Tyramide signaling amplification based immunofluorescence was performed on paraffin-embedded human Testis tissue using Phospho-Rad17 (Ser656) antibody. Antigen retrieval was performed in EDTA buffer pH 9.0 (95 °C, 20 min) followed by cooling to RT. Then incubated with primary antibody (HY-P82895, 1:50 dilution) at room temperature for 60 minutes and HRP conjugated secondary antibody for 10 minutes. Fluorescence was then developed with Vari Fluor 532 TSA (200×)（HY-D1832）. The tissues were counterstained with DAPI and mounted with Anti-fade mounting medium.
Tyramide signaling amplification based immunofluorescence was performed on paraffin-embedded human Testis tissue using Phospho-Rad17 (Ser656) antibody. Antigen retrieval was performed in EDTA buffer pH 9.0 (95 °C, 20 min) followed by cooling to RT. Then incubated with primary antibody (HY-P82895, 1:50 dilution) at room temperature for 60 minutes and HRP conjugated secondary antibody for 10 minutes. Fluorescence was then developed with Vari Fluor 532 TSA (200×)（HY-D1832）. The tissues were counterstained with DAPI and mounted with Anti-fade mounting medium.

Background

Function：Essential for sustained cell growth, maintenance of chromosomal stability, and ATR-dependent checkpoint activation upon DNA damage (PubMed:10208430, PubMed:11418864, PubMed:11687627, PubMed:11799063, PubMed:12672690, PubMed:14624239, PubMed:15235112). Has a weak ATPase activity required for binding to chromatin (PubMed:10208430, PubMed:11418864, PubMed:11687627, PubMed:11799063, PubMed:12672690, PubMed:14624239, PubMed:15235112). Participates in the recruitment of the 9-1-1 (RAD1-RAD9-HUS1) complex and RHNO1 onto chromatin, and in CHEK1 activation (PubMed:21659603). Involved in homologous recombination by mediating recruitment of the MRN complex to DNA damage sites (PubMed:24534091). May also serve as a sensor of DNA replication progression (PubMed:12578958, PubMed:14500819, PubMed:15538388)

Subcellular Localization：Nucleus; Chromosome

Expression：
Tissue_specificity:Overexpressed in various cancer cell lines and in colon carcinoma (at protein level) . Isoform 2 and isoform 3 are the most abundant isoforms in non irradiated cells (at protein level) . Ubiquitous at low levels. Highly expressed in testis, where it is expressed within the germinal epithelium of the seminiferous tubuli. Weakly expressed in seminomas (testicular tumors)

Induction:Induced by X-ray irradiation; Induced by X-ray irradiation; Induced by X-ray irradiation

Isoforms & Post-Translational Modification：O75943 has 4 isomers: O75943-1: 77055 Da (predicted); O75943-2: 75836 Da (predicted); O75943-3: 57244 Da (predicted); O75943-4: 66156 Da (predicted).
Phosphorylation on Ser-646 and Ser-656 is cell cycle-regulated, enhanced by genotoxic stress, and required for activation of checkpoint signaling (PubMed:11418864, PubMed:14500819, PubMed:14871926). Phosphorylation is mediated by ATR upon UV or replication arrest, whereas it may be mediated both by ATR and ATM upon ionizing radiation (PubMed:11418864, PubMed:11799063). Phosphorylation on both sites is required for interaction with RAD1 but dispensable for interaction with RFC3 or RFC4 (PubMed:11687627). Phosphorylation at Thr-633 by ATM in response to DNA damage promotes interaction with NBN and recruitment of the MRN complex to DNA damage sites (PubMed:24534091)

Subunit：Part of a DNA-binding complex containing RFC2, RFC3, RFC4 and RFC5 (PubMed:10884395, PubMed:11572977, PubMed:11687627, PubMed:12400013, PubMed:14624239). Interacts with RAD1 and RAD9 within the 9-1-1 (RAD1-RAD9-HUS1) complex (PubMed:10884395, PubMed:11418864, PubMed:12578958, PubMed:36841485, PubMed:9660800). Interacts with RAD9B, POLE, SNU13 and MCM7 (PubMed:10593953, PubMed:14500819, PubMed:14611806, PubMed:15538388). DNA damage promotes interaction with ATR or ATM and disrupts interaction with the 9-1-1 (RAD1-RAD9-HUS1) complex (PubMed:10884395, PubMed:11418864, PubMed:12578958). Interacts (when phosphorylated) with NBN; promoting recruitment of the MRN complex to DNA damage sites (PubMed:24534091)

Database

Entrez Gene: 5884 Human

SwissProt: O75943 Human

OMIM: 603139 Human

Research Field

Cell Biology

Synonyms

RAD17; R24L; Cell cycle checkpoint protein RAD17; hRad17; RF-C/activator 1 homolog

Documentation

Select Batch:

Data Sheet (235 KB) SDS (252 KB)

COA (206 KB)

User Guide for Antibodies (1077 KB)

Phospho-Rad17 (Ser656) Antibody (YA2640) Related Classifications

Help & FAQs

Do most proteins show cross-species activity?
Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

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Phospho-Rad17 (Ser656) Antibody (YA2640)

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