Revision 7

#9389Store 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, IP, IHC-P, FC-FP, ChIP, ChIP-seq, C&R

REACTIVITY:

H Mk

SENSITIVITY:

Endogenous

MW (kDa):

90

Source/Isotype:

Rabbit IgG

UniProt ID:

#P08047

Entrez-Gene Id:

6667

Product Information

Product Usage Information

For optimal ChIP and ChIP-seq results, use 5 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.
Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Immunohistochemistry (Paraffin) 1:1000 - 1:4000
Flow Cytometry (Fixed/Permeabilized) 1:400 - 1:1600
Chromatin IP 1:100
Chromatin IP-seq 1:100
CUT&RUN 1:50

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #84386.

Specificity / Sensitivity

SP1 (D4C3) Rabbit mAb recognizes endogenous levels of total SP1 protein. It is predicted to detect all three known isoforms.

Species Reactivity:

Human, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro593 of human SP1 protein (Isoform 1).

Background

Specificity protein 1 (SP1) is a ubiquitously expressed transcription factor belonging to the family of C2H2-type zinc finger containing DNA-binding proteins. SP1 binds GC-rich motifs with high affinity and regulates the expression of numerous mammalian genes (1,2). It interacts with many other transcription factors, such as c-Myc, EGR1, and Stat1, and with basal transcription machinery components. SP1 interacts with chromatin-modifying factors, such as histone deacetylases (HDACs) and p300 in chromatin remodeling. Transcriptional activity and stability of SP1 are regulated by post-translational modification, including phosphorylation, acetylation, ubiquitination, and glycosylation (3). Glycosylation of SP1 following insulin treatment leads to increased nuclear localization, while glucagon treatment increases cytoplasmic SP1 levels (4-6). Investigators have found high levels of SP1 in patients with Alzheimer's disease (7).

  1. Kadonaga, J.T. et al. (1987) Cell 51, 1079-90.
  2. Song, J. et al. (2003) Int J Mol Med 11, 547-53.
  3. Tan, N.Y. and Khachigian, L.M. (2009) Mol Cell Biol 29, 2483-8.
  4. Majumdar, G. et al. (2003) Am J Physiol Endocrinol Metab 285, E584-91.
  5. Majumdar, G. et al. (2006) J Biol Chem 281, 3642-50.
  6. Solomon, S.S. et al. (2008) Life Sci 83, 305-12.
  7. Citron, B.A. et al. (2008) J Neurosci Res 86, 2499-504.

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 IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) FC-FP: Flow Cytometry (Fixed/Permeabilized) ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq C&R: CUT&RUN

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.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
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 7
#9389

SP1 (D4C3) Rabbit mAb

Western Blotting Image 1: SP1 (D4C3) Rabbit mAb Expand Image
使用 SP1 (D4C3) Rabbit mAb,对不同细胞系的提取物进行蛋白质印迹分析。
No image available
Immunohistochemistry Image 1: SP1 (D4C3) Rabbit mAb Expand Image
使用 SP1 (D4C3) Rabbit mAb, 对石蜡包埋的人肺癌组织进行免疫组织化学分析。
Immunohistochemistry Image 2: SP1 (D4C3) Rabbit mAb Expand Image
在对照肽(左图)或抗原特异性肽(右图)存在的情况下,使用 SP1 (D4C3) Rabbit mAb 对石蜡包埋的人结肠癌组织进行免疫组织化学分析。
Immunohistochemistry Image 3: SP1 (D4C3) Rabbit mAb Expand Image
使用 SP1 (D4C3) Rabbit mAb 对石蜡包埋的人前列腺癌组织进行免疫组织化学分析。
Flow Cytometry Image 1: SP1 (D4C3) Rabbit mAb Expand Image
以浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)作为对照,使用 SP1 (D4C3) Rabbit mAb(实线)对 Hela 细胞进行流式细胞分析。Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 用作二抗。
Chromatin Immunoprecipitation Image 1: SP1 (D4C3) Rabbit mAb Expand Image
使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9005,对 293 细胞中提取的交联染色质,在加入 SP1 (D4C3) Rabbit mAb 后,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。图形显示了一种已知的 SP1 靶基因 TINF2/TIN2 之间的结合(参阅包含 ChIP-qPCR 数据的其他图)。
Chromatin Immunoprecipitation Image 2: SP1 (D4C3) Rabbit mAb Expand Image
使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9005,对 293 细胞中提取的交联染色质,在加入 SP1 (D4C3) Rabbit mAb 后,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。结果图显示在染色体14(上图)内的结合,包括 TINF2/TIN2 的已知靶基因 SP1(下图)(参见包含 ChIP-qPCR 数据的其他结果图)。
Chromatin Immunoprecipitation Image 3: SP1 (D4C3) Rabbit mAb Expand Image
使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003 对 293 细胞的交联染色质,在加入 SP1 (D4C3) Rabbit mAb 或 Normal Rabbit IgG #2729 进行染色质免疫沉淀。使用 SimpleChIP® Human DHFR Intron 1 Primers #7531、human TIN2 intron 1 primers 以及 SimpleChIP® Human α Satellite Repeat Primers #4486,通过实时 PCR 对富集的 DNA 进行定量分析。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: SP1 (D4C3) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652,以 293 细胞和 SP1 (D4C3) Rabbit mAb 进行 CUT&RUN。使用 DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。该图显示结合作用遍及STK11IP 基因(SP1 的已知靶标基因)(参见包含 CUT&RUN-qPCR 数据的其他结果图)。
CUT and RUN Image 2: SP1 (D4C3) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652,以 293 细胞和 SP1 (D4C3) Rabbit mAb 进行 CUT&RUN。使用 DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。该图显示结合作用遍及染色体 2(上图),包括 SP1 的已知靶标基因 STK11IP(下图)(参见包含 CUT&RUN-qPCR 数据的其他结果图)。
CUT and RUN Image 3: SP1 (D4C3) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652,以 293 细胞和 SP1 (D4C3) Rabbit mAb 或 Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362 进行 CUT&RUN 。使用人 STK11IP 启动子引物和人 FAT4 启动子引物,通过实时 PCR 对富集的 DNA 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。