SP1 Recombinant Rabbit Monoclonal Antibody [JF0950] - BSA and Azide free
Catalog# HA750330
SP1 Recombinant Rabbit Monoclonal Antibody [JF0950] - BSA and Azide free
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WB
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IF-Cell
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IF-Tissue
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IHC-P
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FC
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ChIP
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IP
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Human
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Mouse
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Rat
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ET1702-02
含抗保成分
-
unconjugated
概述
产品名称
SP1 Recombinant Rabbit Monoclonal Antibody [JF0950] - BSA and Azide free
抗体类型
Recombinant Rabbit monoclonal Antibody
免疫原
Synthetic peptide within Human SP1 aa 544-588 / 785.
种属反应性
Human, Mouse, Rat
验证应用
WB, IF-Cell, IF-Tissue, IHC-P, FC, ChIP, IP
分子量
Predicted band size: 81 kDa
阳性对照
HeLa cell lysate, 293T cell lysate, Jurkat cell lysate, U-2 OS cell lysate, COS-1 cell lysate, HeLa, human breast cancer tissue, human colon cancer tissue, human lung tissue, mouse lung tissue, rat lung tissue, MCF-7.
偶联
unconjugated
克隆号
JF0950
产品特性
形态
Liquid
浓度
存放说明
Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
存储缓冲液
1*PBS (pH7.4).
亚型
IgG
纯化方式
Protein A affinity purified.
应用稀释度
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WB
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1:1,000-1:2,000
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IF-Cell
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1:100-1:500
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IF-Tissue
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1:100-1:500
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IHC-P
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1:200-1:1,000
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FC
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1:500-1:1,000
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ChIP
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Use 0.5~2 μg for 25 μg of chromatin.
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IP
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1-2μg/sample
靶点
功能
Sp1 is a sequence-specific transcription factor that recognizes GGGGCGGGGC and closely related sequences, which are often referred to as GC boxes. Sp1 was initially identified as a HeLa cell-derived factor that selectively activates in vitro transcription from the SV40 promoter and binds to the multiple GC boxes in the 21-bp repeated elements in SV40. The sequence specificity of DNA binding is conferred by Zn (II) fingers, whereas a different region of Sp1 appears to regulate the affinity of DNA binding. Sp1 belongs to a subgroup of transcription factors that are phosphorylated upon binding to promoter sequences. Evidence suggests that the early growth response gene, Erg-1 (also known as Zif268 or NGF1-A), may downregulate certain mammalian gene promoters by competing with Sp1 for binding to an overlapping binding motif. The gene encoding human Sp1 maps to chromosome 12q13.1.
背景文献
1. Guaraldo M et al. Characterization of human mitochondrial ferritin promoter: identification of transcription factors and evidences of epigenetic control. Sci Rep 6:33432 (2016).
2. Deng X et al. Protein arginine methyltransferase 5 functions as an epigenetic activator of the androgen receptor to promote prostate cancer cell growth. Oncogene:1-9 (2016).
序列相似性
Belongs to the Sp1 C2H2-type zinc-finger protein family.
组织特异性
Up-regulated in adenocarcinomas of the stomach (at protein level). Isoform 3 is ubiquitously expressed at low levels.
翻译后修饰
Phosphorylated on multiple serine and threonine residues. Phosphorylation is coupled to ubiquitination, sumoylation and proteolytic processing. Phosphorylation on Ser-59 enhances proteolytic cleavage. Phosphorylation on Ser-7 enhances ubiquitination and protein degradation. Hyperphosphorylation on Ser-101 in response to DNA damage has no effect on transcriptional activity. MAPK1/MAPK3-mediated phosphorylation on Thr-453 and Thr-739 enhances VEGF transcription but, represses FGF2-triggered PDGFR-alpha transcription. Also implicated in the repression of RECK by ERBB2. Hyperphosphorylated on Thr-278 and Thr-739 during mitosis by MAPK8 shielding SP1 from degradation by the ubiquitin-dependent pathway. Phosphorylated in the zinc-finger domain by calmodulin-activated PKCzeta. Phosphorylation on Ser-641 by PKCzeta is critical for TSA-activated LHR gene expression through release of its repressor, p107. Phosphorylation on Thr-668, Ser-670 and Thr-681 is stimulated by angiotensin II via the AT1 receptor inducing increased binding to the PDGF-D promoter. This phosphorylation is increased in injured artey wall. Ser-59 and Thr-681 can both be dephosphorylated by PP2A during cell-cycle interphase. Dephosphorylation on Ser-59 leads to increased chromatin association during interphase and increases the transcriptional activity. On insulin stimulation, sequentially glycosylated and phosphorylated on several C-terminal serine and threonine residues.; Acetylated. Acetylation/deacetylation events affect transcriptional activity. Deacetylation leads to an increase in the expression the 12(s)-lipooxygenase gene though recruitment of p300 to the promoter.; Ubiquitinated. Ubiquitination occurs on the C-terminal proteolytically-cleaved peptide and is triggered by phosphorylation.; Sumoylated with SUMO1. Sumoylation modulates proteolytic cleavage of the N-terminal repressor domain. Sumoylation levels are attenuated during tumorigenesis. Phosphorylation mediates SP1 desumoylation.; Proteolytic cleavage in the N-terminal repressor domain is prevented by sumoylation. The C-terminal cleaved product is susceptible to degradation.; O-glycosylated; Contains 8 N-acetylglucosamine side chains. Levels are controlled by insulin and the SP1 phosphorylation states. Insulin-mediated O-glycosylation locates SP1 to the nucleus, where it is sequentially deglycosylated and phosphorylated. O-glycosylation affects transcriptional activity through disrupting the interaction with a number of transcription factors including ELF1 and NFYA. Also inhibits interaction with the HIV1 promoter. Inhibited by peroxisomome proliferator receptor gamma (PPARgamma).
亚细胞定位
Nucleus, Cytoplasm.
别名
SP 1 antibody
SP1 antibody
Sp1 transcription factor antibody
SP1_HUMAN antibody
Specificity protein 1 antibody
Transcription factor Sp1 antibody
TSFP 1 antibody
TSFP1 antibody
图片
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Western blot analysis of SP1 on different lysates with Rabbit anti-SP1 antibody (HA750330) at 1/1,000 dilution.
Lane 1: HeLa cell lysate
Lane 2: 293T cell lysate
Lane 3: Jurkat cell lysate
Lane 4: U-2 OS cell lysate
Lane 5: COS-1 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 81 kDa
Observed band size: 81 kDa
Exposure time: 4 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750330) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunocytochemistry analysis of HeLa cells labeling SP1 with Rabbit anti-SP1 antibody (HA750330) at 1/200 dilution.
Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-SP1 antibody (HA750330) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-SP1 antibody (HA750330) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750330) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-SP1 antibody (HA750330) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750330) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human lung tissue with Rabbit anti-SP1 antibody (HA750330) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750330) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse lung tissue with Rabbit anti-SP1 antibody (HA750330) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750330) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat lung tissue with Rabbit anti-SP1 antibody (HA750330) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750330) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Flow cytometric analysis of MCF-7 cells labeling SP1.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA750330, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells with SP1 (HA750330) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. 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.
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SP1 was immunoprecipitated from 0.2 mg HeLa cell lysate with HA750330 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA750330 at 1/1,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: HeLa cell lysate (input)
Lane 2: HA750330 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of HA750330 in HeLa cell lysate
Blocking/Dilution buffer: primary antibody dilution (K1803)
Exposure time: 6 seconds; ECL: K1801
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