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#25501 SET1/COMPASS Antibody Sampler Kit

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希望納入価格 (円)
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2019年1月17日15時10分 現在
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キット内容 容量 用途 種交差性 ホモロジー† 検出分子量 アイソタイプ
SET1A (D3V9S) Rabbit mAb #61702 20 µl WB, IP, ChIP, ChIP-seq H 300 Rabbit IgG
SET1B (D1U5D) Rabbit mAb #44922 20 µl WB, IP H, M, Mk B, Pg, Hr 320 Rabbit IgG
MLL1 (D2M7U) Rabbit mAb (Amino-terminal Antigen) #14689 20 µl WB, IP H, M, R, Mk 300 Rabbit IgG
MLL1 (D6G8N) Rabbit mAb (Carboxy-terminal Antigen) #14197 20 µl WB, IP, IF-IC H, M, R, Mk 180 Rabbit IgG
MLL2/KMT2B (D6X2E) Rabbit mAb (Carboxy-terminal Antigen) #63735 20 µl WB, IP, IF-IC H 80 Rabbit IgG
WDR5 (D9E1I) Rabbit mAb #13105 20 µl WB, ChIP, ChIP-seq H, M, R, Mk 37 Rabbit IgG
WDR82 (D2I3B) Rabbit mAb #99715 20 µl WB, IP H, M, Mk Hm, Hr 30 Rabbit IgG
Menin (D45B1) XP® Rabbit mAb #6891 20 µl WB, IF-IC H, M, R, Mk B, Pg, Hr 68 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl WB Goat

†Species predicted to react based on 100% sequence homology.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, ChIP=Chromatin IP, ChIP-seq=Chromatin IP-seq, IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key: H=Human, M=Mouse, Mk=Monkey, R=Rat

貯法
-20℃
社内データ

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 various cell lines using Menin (D45B1) XP® Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using WDR5 (D9E1I) Rabbit mAb.


Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using MLL1 (D2M7U) Rabbit mAb (Amino-terminal Antigen).

Western Blotting

Western Blotting

Western blot analysis of extracts from MLL1 wild-type (WT) and knockout (KO) mouse embryonic fibroblasts (MEF) using MLL1 (D6G8N) Rabbit mAb (Carboxy-terminal Antigen) (upper) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower). MLL1 WT and KO MEF were kindly provided by Dr. Ali Shilatifard of Northwestern University.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T and HeLa cells using SET1A (D3V9S) Rabbit mAb.


Western Blotting

Western Blotting

Western blot analysis of extracts from HCT 116, NIH/3T3, and COS-7 cell lines using WDR82 (D2I3B) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using MLL2/KMT2B (D6X2E) Rabbit mAb (Carboxy-terminal Antigen) (upper) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower). As expected, HCT 116 cells are negative for MLL2 expression.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T, F9, and COS-7 cell lines using SET1B (D1U5D) Rabbit mAb.


Chromatin IP-seq

Chromatin IP-seq

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and SET1A (D3V9S) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across GAPDH, a known target gene of SET1A (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.

Chromatin IP-seq

Chromatin IP-seq

Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and WDR5 (D9E1I) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across NANOG, a known target gene of WDR5 (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.

IF-IC

IF-IC

Confocal immunofluorescent analysis of COS-7 cells using Menin (D45B1) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).


Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and either WDR5 (D9E1I) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using human nanog promoter primers, SimpleChIP® Human Oct-4 Promoter Primers #4641, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using MLL1 (D6G8N) Rabbit mAb (Carboxy-terminal Antigen).

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either SET1A (D3V9S) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by Real-Time PCR using SimpleChIP® Human β-Actin Promoter Primers #13653, SimpleChIP® Human GAPDH Promoter Primers #4471, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.


IP

IP

Immunoprecipitation of SET1A from 293T cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is SET1A (D3V9S) Rabbit mAb. Western blot analysis was performed using SET1A (D3V9S) Rabbit mAb.

IP

IP

Immunoprecipitation of MLL2/KMT2B from NCCIT cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is MLL2/KMT2B (D6X2E) Rabbit mAb (Carboxy-terminal Antigen). Western blot analysis was performed using MLL2/KMT2B (D6X2E) Rabbit mAb (Carboxy-terminal Antigen).

IP

IP

Immunoprecipitation of SET1B from HeLa cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is SET1B (D1U5D) Rabbit mAb. Western blot analysis was performed using SET1B (D1U5D) Rabbit mAb.


IF-IC

IF-IC

Confocal immunofluorescent analysis of MEF wild-type (left, positive) and MEF MLL (-/-) (right, negative) cells using MLL (D6G8N) Rabbit mAb (Carboxy-terminal Antigen) (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). MLL1 WT and KO MEF were kindly provided by Dr. Ali Shilatifard of Northwestern University.

IF-IC

IF-IC

Confocal immunofluorescent analysis of A549 cells (left, positive) and HCT 116 cells (right, negative) using MLL2/KMT2B (D6X2E) Rabbit mAb (Carboxy-terminal Antigen) (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red).

バックグラウンド

The Set1 histone methyltransferase protein was first identified in yeast as part of the Set1/COMPASS histone methyltransferase complex, which methylates histone H3 on lysine 4 and functions as a transcriptional co-activator (1). While yeast contain only one known Set1 protein, mammals contain six Set1-related proteins: SET1A, SET1B, MLL1, MLL2, MLL3 and MLL4, all of which methylate histone H3 on lysine 4 (2,3). These Set1-related proteins are each found in distinct protein complexes, all of which share the common core structural subunits WDR5, RBBP5 and ASH2L (2-6). WDR82 is a core subunit specific to SET1A and SET1B complexes, while Menin is a core subunit specific to the MLL complexes (4,5,7).

Like yeast Set1, all six Set1-related mammalian proteins methylate histone H3 on lysine 4 (2-6). SET1A, SET1B, MLL1 and MLL2 mediate di- and tri-methylation of histone H3 Lys4 at gene promoters to facilitate transcription activation. MLL3 and MLL4 function primarily to mono-methylate histone H3 Lys4 at gene enhancers. MLL1 and MLL2 function as master regulators of both embryogenesis and hematopoiesis, and are required for proper expression of Hox genes (8-10). MLL1 is a large approximately 4000 amino acid protein that is cleaved by the Taspase 1 threonine endopeptidase to form N-terminal (MLL1-N) and C-terminal MLL1 (MLL1-C) fragments, both of which are subunits of the functional MLL1/COMPASS complex (11,12). MLL1 translocations are found in a large number of hematological malignancies, suggesting that Set1 histone methyltransferase complexes play a critical role in leukemogenesis (6). Like MLL1, MLL2 is also a large, approximately 2700 amino acid protein that is cleaved by the Taspase 1 threonine endopeptidase to form N-terminal (MLL2-N) and C-terminal (MLL2-C) fragments, both of which are subunits of the functional MLL2/COMPASS complex. MLL2 has also been implicated as a modulator of hematological malignancies (13). MLL3 and MLL4 proteins are not cleaved by Taspase 1.

使用例
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関連製品
13105   WDR5 (D9E1I) Rabbit mAb
14197   MLL1 (D6G8N) Rabbit mAb (Carboxy-terminal Antigen)
14689   MLL1 (D2M7U) Rabbit mAb (Amino-terminal Antigen)
44922   SET1B (D1U5D) Rabbit mAb
61702   SET1A (D3V9S) Rabbit mAb
63735   MLL2/KMT2B (D6X2E) Rabbit mAb (Carboxy-terminal Antigen)
6891   Menin (D45B1) XP® Rabbit mAb
7003   20X LumiGLO® Reagent and 20X Peroxide
7071   Phototope-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
7074   Anti-rabbit IgG, HRP-linked Antibody
7727   Biotinylated Protein Ladder Detection Pack
99715   WDR82 (D2I3B) Rabbit mAb
9999   Nonfat Dry Milk

Illumina is a registered trademark of Illumina, Inc.
Tween is a registered trademark of ICI Americas, Inc.
DRAQ5 is a registered trademark of Biostatus Limited.
XP is a registered trademark of Cell Signaling Technology, Inc.
SimpleChIP 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.
DyLight is a trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.

本製品は試験研究用です。

SET1/COMPASS Antibody Sampler Kit

Immune Cell Signaling Pathways

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