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#9931 Phospho-Chk1/2 Antibody Sampler Kit

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2019年3月20日11時40分 現在
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#9931T1 Kit115,000
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キット内容 容量 用途 種交差性 検出分子量 アイソタイプ
Phospho-Chk1 (Ser317) (D12H3) XP® Rabbit mAb #12302 20 µl WB, IP, IF-IC H, M, Mk 56 Rabbit IgG
Phospho-Chk1 (Ser345) (133D3) Rabbit mAb #2348 20 µl WB, IF-IC, F H, M, R, Mk 56 Rabbit IgG
Phospho-Chk1 (Ser296) Antibody #2349 20 µl WB H, Mk 56 Rabbit
Chk1 (2G1D5) Mouse mAb #2360 20 µl WB H, M, R, Mk 56 Mouse IgG1
Chk2 (D9C6) XP® Rabbit mAb #6334 20 µl WB, IP, IHC-P, IF-IC H 62 Rabbit IgG
Phospho-Chk2 (Ser19) Antibody #2666 20 µl WB, IHC-P H 62 Rabbit
Phospho-Chk2 (Ser33/35) Antibody #2665 20 µl WB H, Mk 62 Rabbit
Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb #2197 20 µl WB, IP, IHC-P, F H 62 Rabbit IgG
Phospho-Chk2 (Ser516) Antibody #2669 20 µl WB H 62 Rabbit
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl WB Goat

Applications Key: W=Western Blotting, IP=Immunoprecipitation, IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry, IHC-P=Immunohistochemistry (Paraffin)
Reactivity Key: H=Human, M=Mouse, Mk=Monkey, R=Rat

貯法
-20℃
※括弧付きの動物種は配列が100%相同であるため反応すると推定されます。
社内データ

Western blot analysis of extracts from Hela and Cos cells, untreated or treated with 100 mJ/cm2 UV light with 1 hour recovery, using Phospho-Chk1 (Ser296) Antibody, #2349.

Western Blotting

Western Blotting

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

Western Blotting

Western Blotting

Western blot analysis of extracts from COS cells, untransfected (lane 1) or transfected with Wild-type Chk2 (lane 2), Chk2 (S19A) (lane 3), Chk2 (T26S28A) (lane 4), Chk2 (S33S35A) (lane 5) or Chk2 (T68A) (lane 6), using Phospho-Chk2 (Ser33/35) Antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from COS cells, untransfected (lane 1) or transfected with Wild-type Chk2 (lane 2), Chk2 (S19A) (lane 3), Chk2 (T26S28A) (lane 4), Chk2 (S33S35A) (lane 5) or Chk2 (T68A) (lane 6), using Phospho-Chk2 (Ser19) Antibody.


Western Blotting

Western Blotting

Western blot analysis of extracts from 293 cells, untreated or UV-treated (50 mJ/cm2, 2hrs), using Phospho-Chk2 (Ser516) Antibody (upper) or Chk2 Antibody #2662 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa, COS, NIH/3T3 and C6 cells, untreated or UV-treated, using Phospho-Chk1 (Ser345) (133D30) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from Hela and Cos cells, untreated or treated with 100 mJ/cm2 UV light with 1 hour recovery, using Phospho-Chk1 (Ser296) Antibody.


Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Chk1 (2G1D5) Mouse mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, untreated or UV-treated, using Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Chk2 (D9C6) XP® Rabbit mAb.


Western Blotting

Western Blotting

Western blot analysis of extracts from 293 and NIH/3T3 cells, untreated (-) or UV-treated (100 mJ, 1 hr recovery; +), using Phospho-Chk1 (Ser317) (D12H3) XP® Rabbit mAb. The blot on the right was treated with calf intestinal phosphatase (CIP) before western blot.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells treated with UV for the indicated times, using Phospho-Chk2 (Ser33/35) Antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells treated with UV for the indicated times, using Phospho-Chk2 (Ser19) Antibody.


IP

IP

Immunoprecipitation of phospho-chk2 from UV-treated HT29 cells using Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb followed by western blot using the same antibody.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HeLa cells, untreated (blue) and UV-treated (green), using Phospho-Chk1 (Ser345) (133D3) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® Chk1 siRNA I #6241 or SignalSilence® Chk1 siRNA II (+), using Chk1 (2G1D5) Mouse mAb #2360 and β-Actin (13E5) Rabbit mAb #4970. Chk1 (2G1D5) Mouse mAb confirms silencing of Chk1 expression and β-Actin (13E5) Rabbit mAb is used to control for loading and specificity of Chk1 siRNA.


IP

IP

Immunoprecipitation of Chk2 from 293 cell extracts using Chk2 (D9C6) XP® Rabbit mAb (lane 2). Western blot detection was performed using the same antibody. Lane 1 is 10% input.

IP

IP

Immunoprecipitation of phospho-Chk1 (Ser317) from 293 cell extracts treated with UV (100 mJ, 1 hr recovery) using Phospho-Chk1 (Ser317) (D12H3) XP® Rabbit mAb (lane 2) or Rabbit (D1AG) mAb IgG XP® Isotype Control #3900 (lane 3). Lane 1 is 10% input.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, showing nuclear localization, using Phospho-Chk2 (Ser19) Antibody.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of C2C12 cells, untreated (left) or UV-treated (right), using Phospho-Chk1 (Ser345) (133D3) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcioma using Chk2 (D9C6) XP® Rabbit mAb.


IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left), UV-treated (center), or UV and λ phosphatase-treated (right), using Phospho-Chk1 (Ser317) (D12H3) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, control (left) or λ phosphatase-treated (right), using Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human ovarian serous adenocarcinoma using Chk2 (D9C6) XP® Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded cell pellets, HCT 116 (left) or MDA-MB-231 (right), using Chk2 (D9C6) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded HT-29 cell pellets, control (left) or UV-treated (right), using Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb.


IF-IC

IF-IC

Confocal immunofluorescent analysis of HCT 116 (left) and HCT-15 (right) cells using Chk2 (D9C6) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of untreated Jurkat cells, using Phospho-Chk2 (Thr68) (C13C1) Rb mAb versus propidium iodide (DNA content). The boxed population indicates phospho-Chk2 (Thr68)-positive cells.

バックグラウンド

Chk1 kinase acts downstream of ATM/ATR kinase and plays an important role in DNA damage checkpoint control, embryonic development, and tumor suppression (1). Activation of Chk1 involves phosphorylation at Ser317 and Ser345 by ATM/ATR, followed by autophosphorylation of Ser296. Activation occurs in response to blocked DNA replication and certain forms of genotoxic stress (2). While phosphorylation at Ser345 serves to localize Chk1 to the nucleus following checkpoint activation (3), phosphorylation at Ser317 along with site-specific phosphorylation of PTEN allows for re-entry into the cell cycle following stalled DNA replication (4). Chk1 exerts its checkpoint mechanism on the cell cycle, in part, by regulating the cdc25 family of phosphatases. Chk1 phosphorylation of cdc25A targets it for proteolysis and inhibits its activity through 14-3-3 binding (5). Activated Chk1 can inactivate cdc25C via phosphorylation at Ser216, blocking the activation of cdc2 and transition into mitosis (6). Centrosomal Chk1 has been shown to phosphorylate cdc25B and inhibit its activation of CDK1-cyclin B1, thereby abrogating mitotic spindle formation and chromatin condensation (7). Furthermore, Chk1 plays a role in spindle checkpoint function through regulation of aurora B and BubR1 (8). Research studies have implicated Chk1 as a drug target for cancer therapy as its inhibition leads to cell death in many cancer cell lines (9).

Chk2 is the mammalian homologue of the budding yeast Rad53 and fission yeast Cds1 checkpoint kinases (5-7). The amino-terminal domain of Chk2 contains a series of seven serine or threonine residues (Ser19, Thr26, Ser28, Ser33, Ser35, Ser50 and Thr68) followed by glutamine (SQ or TQ motif). These are known to be preferred sites for phosphorylation by ATM/ATR kinases (8). Indeed, after DNA damage by ionizing radiation (IR), UV irradiation and DNA replication blocked by hydroxyurea, Thr68 and other sites in this region become phosphorylated by ATM/ATR (9-11). The SQ/TQ cluster domain, therefore, seems to have a regulatory function. Phosphorylation at Thr68 is a prerequisite for the subsequent activation step, which is attributable to autophosphorylation of Chk2 on residues Thr383 and Thr387 in the activation loop of the kinase domain (12).

使用例
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DRAQ5 is a registered trademark of Biostatus Limited.
XP is a registered trademark of Cell Signaling Technology, Inc.
LumiGLO is a registered trademark of Kirkegaard & Perry Laboratories.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

本製品は試験研究用です。

Phospho-Chk1/2 Antibody Sampler Kit

Immune Cell Signaling Pathways

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