Revision 6

#97800Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IF-IC, ChIP, ChIP-seq, C&R, C&T

REACTIVITY:

H M Mk

SENSITIVITY:

Endogenous

MW (kDa):

50-75

Source/Isotype:

Rabbit IgG

UniProt ID:

#O75030

Entrez-Gene Id:

4286

Product Information

Product Usage Information

For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.

The CUT&Tag dilution was determined using CUT&Tag Assay Kit #77552.

Application Dilution
Western Blotting 1:1000
Immunofluorescence (Immunocytochemistry) 1:100 - 1:200
Chromatin IP 1:50
Chromatin IP-seq 1:50
CUT&RUN 1:50
CUT&Tag 1:50

Specificity / Sensitivity

MITF (D3B4T) Rabbit mAb recognizes endogenous levels of total MITF protein.

Species Reactivity:

Human, Mouse, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the carboxy terminus of human MITF protein.

Background

Microphthalmia-associated transcription factor (MITF) is a basic helix-loop-helix leucine zipper transcription factor that is most widely known for its roles in melanocyte, ophthalmic, and osteoclast development (1-3). In humans, MITF can function as a melanoma oncogene (4) and mutations in the corresponding MITF gene are associated with Waardenburg syndrome type 2, an auditory-pigmentary syndrome characterized by developmental defects in cells derived from neural crest (5). At least 12 isoforms of MITF have been identified, which exhibit differential patterns of expression among cell and tissue types (6).

  1. Hershey, C.L. and Fisher, D.E. (2004) Bone 34, 689-96.
  2. Nomura, S. et al. (2001) J Bone Miner Metab 19, 183-7.
  3. Widlund, H.R. and Fisher, D.E. (2003) Oncogene 22, 3035-41.
  4. Levy, C. et al. (2006) Trends Mol Med 12, 406-14.
  5. Read, A.P. and Newton, V.E. (1997) J Med Genet 34, 656-65.
  6. Tachibana, M. (2000) Pigment Cell Res 13, 230-40.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IF-IC: Immunofluorescence (Immunocytochemistry) ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq C&R: CUT&RUN C&T: CUT&Tag

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Revision 6
#97800

MITF (D3B4T) Rabbit mAb

Western Blotting Image 1: MITF (D3B4T) Rabbit mAb Expand Image
使用 MITF (D3B4T) Rabbit mAb(上)和 β-Actin (D6A8) Rabbit mAb #8457(下),对不同细胞系提取物进行蛋白质印迹分析。
Immunofluorescence Image 1: MITF (D3B4T) Rabbit mAb Expand Image
使用 MITF (D3B4T) Rabbit mAb(绿色)、DyLight 650 Phalloidin #12956(红色)和 DAPI #4083(蓝色)对 SK-MEL-28 细胞(左图,阳性)和 HCT 116 细胞(右图,阴性)进行共聚焦免疫荧光分析。
Chromatin Immunoprecipitation Image 1: MITF (D3B4T) Rabbit mAb Expand Image
使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003,对 SK-MEL-28 细胞的交联染色质和 MITF (D3B4T) Rabbit mAb 进行染色质免疫沉淀。使用 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。结果图显示在 BEST1/VMD2 内结合,BEST1/VMD2 是已知的 MITF 靶标基因(参见包含 ChIP-qPCR 数据的其他结果图)。如需了解其他 ChIP-seq 情况,请下载产品说明书。
Chromatin Immunoprecipitation Image 2: MITF (D3B4T) Rabbit mAb Expand Image
使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003,对 SK-MEL-28 细胞的交联染色质和 10 μl 的 MITF (D3B4T) Rabbit mAb 进行染色质免疫沉淀分析。使用 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。结果图显示在染色体 11(上图)内的结合,包括 MITF 的已知靶标基因 BEST1/VMD2(下图)(参见包含 ChIP-qPCR 数据的其他结果图)。
Chromatin Immunoprecipitation Image 3: MITF (D3B4T) Rabbit mAb Expand Image
使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003,对 SK-MEL-28 细胞中提取的交联染色质,在加入 MITF (D3B4T) Rabbit mAb 或 Normal Rabbit IgG #2729 后,进行染色质免疫沉淀。使用 SimpleChIP® Human Tyrosinase Promoter Primers #12970、Human VMD2 Intron 1 Primers 和 SimpleChIP® Human α Satellite Repeat Primers #4486 并通过实时 PCR 对富集的 DNA 进行定量。将每份样品中免疫沉淀的 DNA 的量表现为相对于所输入染色质总量(等于 1)的信号。
CUT and RUN Image 1: MITF (D3B4T) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652,通过 SM-MEL-28 细胞和 MITF (D3B4T) Rabbit mAb进行 CUT&RUN。‚使用 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。图形显示在 TYR 内结合,TYR是一种已知的 MITF 靶标基因(参见包含 CUT&RUN-qPCR 数据的其他图形)。
CUT and RUN Image 2: MITF (D3B4T) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652,通过 SM-MEL-28 细胞和 MITF (D3B4T) Rabbit mAb进行 CUT&RUN。‚使用 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。结果图显示在染色体 11(上图)内的结合,包括 MITF 的已知靶标基因 TYR(下图)(参见包含 CUT&RUN-qPCR 数据的其他结果图)。
CUT and RUN Image 3: MITF (D3B4T) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652,通过€‚ SM-MEL-28 细胞和 MITF (D3B4T) Rabbit mAb 或 Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362进行 CUT&RUN。使用 SimpleChIP® Human Tyrosinase Promoter Primers #12970 和 SimpleChIP® Human α Satellite Repeat Primers #4486,通过实时 PCR 定量富集的 DNA。将每份样品中免疫沉淀的 DNA 的量表现为相对于所输入染色质总量(等于 1)的信号。
CUT and Tag Image 1: MITF (D3B4T) Rabbit mAb Expand Image
CUT&Tag was performed with SK-MEL-28 cells and MITF (D3B4T) Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figure shows binding across BEST1/VMD2, a known target gene of MITF (see our ChIP-qPCR figure).
CUT and Tag Image 2: MITF (D3B4T) Rabbit mAb Expand Image
CUT&Tag was performed with SK-MEL-28 cells and MITF (D3B4T) Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figures show binding across chromosome 11 (upper), including BEST1/VMD2 (lower), a known target gene of MITF (see our ChIP-qPCR figure).