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StressMarq/Anti-Dnmt3L Antibody [S117-9]/SMC-341D-A633/100-µg
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StressMarq/Anti-Dnmt3L Antibody [S117-9]/SMC-341D-A633/100-µg
品牌 / 
StressMarq
货号 / 
SMC-341D-A633
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Overview:

Product Name Dnmt3L Antibody
Description

Mouse Anti-Human Dnmt3L Monoclonal IgG1

Species Reactivity Human, Mouse
Applications WB, IHC, ICC/IF, IP, AM
Antibody Dilution WB (1:1000), IHC (1:1000), ICC/IF (1:100); optimal dilutions for assays should be determined by the user.
Host Species Mouse
Immunogen Species Human
Immunogen Fusion protein amino acids 1-387 of human DNMT3L
Concentration 1 mg/ml
Conjugates Alkaline Phosphatase, APC, ATTO 390, ATTO 488, ATTO 565, ATTO 594, ATTO 633, ATTO 655, ATTO 680, ATTO 700, Biotin, FITC, HRP, PE/ATTO 594, PerCP, RPE, Streptavidin, Unconjugated

Properties

Storage Buffer PBS pH7.4, 50% glycerol, 0.09% sodium azide
Storage Temperature -20ºC
Shipping Temperature Blue Ice or 4ºC
Purification Protein G Purified
Clonality Monoclonal
Clone Number S117-9
Isotype IgG1
Specificity Detects ~45kDa. No cross-reactivity against DNMT3a or DNMT3B.
Cite This Product StressMarq Biosciences Cat# SMC-341, RRID: AB_2094142
Certificate of Analysis 1 µg/ml of SMC-341 was sufficient for detection of DNMT3L in 10 µg of COS cell transiently transfected with Flag-tagged DNMT3L lysate by colorimetric immunoblot analysis using goat anti-mouse IgG:HRP as the secondary antibody.

Biological Description

Alternative Names Cytosine 5 methyltransferase 3 like protein antibody, DNA (cytosine 5 ) methyltransferase 3 like antibody, DNA (cytosine-5)-methyltransferase 3-like antibody, DNA cytosine 5 methyltransferase 3 like protein antibody, DNA methyltransferase 3 like protein antibody, DNM3L_HUMAN antibody, Dnmt 3L antibody, Dnmt3l antibody, Human cytosine 5 methyltransferase 3 like protein antibody, MGC1090 antibody
Research Areas Cell Signaling, Epigenetics, Epigenetics and Nuclear Signaling
Cellular Localization Nucleus
Accession Number NP_037501.2
Gene ID 29947
Swiss Prot Q9UJW3
Scientific Background Methylation of DNA at cytosine residues plays an important role in regulation of gene expression, genomic imprinting and is essential for mammalian development. Hypermethylation of CpG islands in tumor suppressor genes or hypomethylation of bulk genomic DNA may be linked with development of cancer. To date, 3 families of mammalian DNA methyltransferase genes have been identified which include Dnmt1, Dnmt2 and Dnmt3. Dnmt1 is constitutively expressed in proliferating cells and inactivation of this gene causes global demethylation of genomic DNA and embryonic lethality. Dnmt2 is expressed at low levels in adult tissues and its inactivation does not affect DNA methylation or maintenance of methylation. The Dnmt3 family members, Dnmt3a and Dnmt3b, are strongly expressed in ES cells but their expression is down regulated in differentiating ES cells and is low in adult somatic tissue (1-6). Studies show that DNMT3L regulates the activity of DNMT3A and DNMT3B and stimulates their catalytic activities (7). DNMT3L has specifically been linked to the process of carcinogenesis, thru its role in nuclear programming (8).
References 1. Bestor T., et al.(1988) J.Mol. Biol. 203: 971-983.
2. Yen R.W., Vertino P.M., Nelkin B.D. et al. (1992) Nucl. Acids Res. 20: 2287-2291.
3. Xie S. et al. (1999) Gene 236: 87-95.
4. Okano M., Bell D.W., Haber D.A. and Li E. (1999) Cell 99: 247-257.
5. Reik W. et al. (1999) J. Nat. Genet 23: 380-382.
6. Robertson K.D., et al. (2000) Nat Genet 25(3):338-342.
7. Kareta MS., et al. (2006) J Biol Chem. 281(36): 25893-25902.
8. Gokul G., Ramakrishna G., and Khosla S. (2009) Epigenetics. 4(5): 322-329.

Product Images

<p>Immunohistochemistry analysis using Mouse Anti-Dnmt3L Monoclonal Antibody, Clone S117-9 (SMC-341). Tissue: backskin. Species: Mouse. Fixation: Bouin’s Fixative and paraffin-embedded. Primary Antibody: Mouse Anti-Dnmt3L Monoclonal Antibody (SMC-341) at 1:100 for 1 hour at RT. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:50 for 1 hour at RT. Localization: Lower layers of the epidermis have positive cell staining in the cytoplasm.</p>

Immunohistochemistry analysis using Mouse Anti-Dnmt3L Monoclonal Antibody, Clone S117-9 (SMC-341). Tissue: backskin. Species: Mouse. Fixation: Bouin’s Fixative and paraffin-embedded. Primary Antibody: Mouse Anti-Dnmt3L Monoclonal Antibody (SMC-341) at 1:100 for 1 hour at RT. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:50 for 1 hour at RT. Localization: Lower layers of the epidermis have positive cell staining in the cytoplasm.

<p>Western Blot analysis of Human tissue lysate showing detection of Dnmt3L protein using Mouse Anti-Dnmt3L Monoclonal Antibody, Clone S117-9 (SMC-341). Load: 15 µg protein. Block: 1.5% BSA for 30 minutes at RT. Primary Antibody: Mouse Anti-Dnmt3L Monoclonal Antibody (SMC-341) at 1:1000 for 2 hours at RT. Secondary Antibody: Sheep Anti-Mouse IgG: HRP for 1 hour at RT.</p>

Western Blot analysis of Human tissue lysate showing detection of Dnmt3L protein using Mouse Anti-Dnmt3L Monoclonal Antibody, Clone S117-9 (SMC-341). Load: 15 µg protein. Block: 1.5% BSA for 30 minutes at RT. Primary Antibody: Mouse Anti-Dnmt3L Monoclonal Antibody (SMC-341) at 1:1000 for 2 hours at RT. Secondary Antibody: Sheep Anti-Mouse IgG: HRP for 1 hour at RT.

<p>Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-Dnmt3L Monoclonal Antibody, Clone S117-9 (SMC-341). Tissue: HaCaT cells. Species: Human. Fixation: Cold 100% methanol for 10 minutes at -20°C. Primary Antibody: Mouse Anti-Dnmt3L Monoclonal Antibody (SMC-341) at 1:100 for 1 hour at RT. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:50 for 1 hour at RT. Localization: Staining in the nucleus, and accumulated aroudn the nucleus. Clear nuclear demarcations.</p>

Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-Dnmt3L Monoclonal Antibody, Clone S117-9 (SMC-341). Tissue: HaCaT cells. Species: Human. Fixation: Cold 100% methanol for 10 minutes at -20°C. Primary Antibody: Mouse Anti-Dnmt3L Monoclonal Antibody (SMC-341) at 1:100 for 1 hour at RT. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:50 for 1 hour at RT. Localization: Staining in the nucleus, and accumulated aroudn the nucleus. Clear nuclear demarcations.

Product Citations (2)

Other Citations

Biomarker Analysis with Grating Coupled Surface Plasmon Coupled Fluorescence.

Mendoza, A., Dias, J.A., Zeltner, T. and Lawrence, D.A. (2014) J Adv Bio & Biotech. 1(1): 1-22.

PubMed ID: N/A Reactivity Human Applications: Antibody Microarray

Biomarker Analysis with Grating Coupled Surface Plasmon Coupled Fluorescence.

Mendoza, A., Dias, J.A., Zeltner, T. and Lawrence, D.A. (2014) J Adv Bio & Biotech. 1(1): 1-22.

PubMed ID: N/A Reactivity Mouse Applications: Antibody Microarray

 ATTO 633
Overview:

  • High fluorescence yield
  • High thermal and photostability
  • Moderately hydrophilic
  • Good solubility in polar solvents
  • Stable at pH 4 – 11
  • Cationic dye, perchlorate salt
  • Molar Mass: 652.2 g/mol

ATTO 633 Datasheet

ATTO 633 Fluorophore Absorption and Emission SpectrumOptical Properties:

λex = 629 nm

λem = 657 nm

εmax = 1.3×105

Φf = 0.64

τfl = 3.2 ns

Brightness = 83.2

Laser = 633 nm

Filter set = Cy®5

 

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上样缓冲液也有多种,一般是80ul的样本加20ul的上样缓冲液,这样基本上上样50ug的体积在10ul左右。
最好是测浓度,一般不超过30-40ug,上样缓冲液一般4×或者5×,这个没太大区别。
作为指示剂,因为溴酚蓝呈蓝色,而蛋白质是白色,在SDS-凝胶中不明显,且溴酚蓝的相对分子量比蛋白质(绝大部分)小,电泳时速度比蛋白质稍快,因此当溴酚蓝到达电泳槽底部(可看蓝色调带),则电泳结束。
50×TAE Buffer 配制方法:
1。称量氨基丁三醇 242g,Na2EDTA.2H2O 37.2g 于1L烧杯中;
2。向烧杯中加入约600ml去离子水,充分搅拌均匀;
3。加入57.1ml的冰乙酸,充分溶解;
4。用NaOH调pH至8.3,加去离子水定容至1L后,室温保存。
使用时稀释50倍 即1×TAE Buffer
10×TBE Buffer配制方法:
1。称量氨基丁三醇 108g,Na2EDTA.2H2O 7.44g,硼酸55g 于1L烧杯中;
2。加入约700ml去离子水,搅拌均匀;
3。用NaOH调pH至8.3,加去离子水定容至1L后,室温保存。
使用时稀释10倍 即1×TBE Buffer
4×蛋白上样缓冲液使用说明123
songjunlai19922016-03-22

请问有人知道蛋白上样缓冲液应该怎么配制吗?请赐教

蛋白质(经硫酸铵和等电点沉淀后用1MKCL复溶)一加2x或6X上样缓冲液就产生沉淀,试问何故?怎样解决?
经搜索发现类似的问题,但人家的裂解液中含盐酸胍,而我的溶液中是不含含盐酸胍啊。
请做蛋白质的行家帮忙解答,谢谢。
不太好操作,里面有小分子的SDS、β巯基乙醇,较难浓缩,不是有4X的买么,还可以自己配啊,网上都有现成的配方,溴酚蓝、SDS、β巯基乙醇。。。
2ul 5×上样缓冲液中添加3ul水,混匀,即得2×上样缓冲液
(以下内容仅供参考)
2乘SDS-PAGE上样缓冲液(20ml体系)
1.0mol/L Tris-HCL (pH 6.8) 2ml
1.0mol/L DTT 4ml
SDS 0.8g
溴芬蓝 0.04g
甘油 4ml
dd水 定容至20ml
4℃冰箱保存,可以不分装,我们实验室一般用三个月以上
先配1.0mol/L的DTT,双蒸无菌水溶解4℃冰箱保存,配上样缓冲液的时候加入就可以了,都混匀保存也没问题的
提取蛋白,western上样前,加上样缓冲液煮,煮之前是澄清透明的,煮完就有挺多的沉淀生成,不知道什么原因,怎么解决,求帮忙!
WB试剂配方123
异鸣23692021-07-25
10 mmol/L Tris-HCl(pH 8.0) 1 mmol/L EDTA(pH 8.0) 因为含有以上两种物质,所以称为TE。
配制分三步: 1) 1 M Tris-HCl (pH 8.0) 50 ml的配制:称取Tris碱6.06 g,加超纯水40 ml溶解,滴加浓HCl约2.1 ml调pH至8.0,定容至50 ml。
上样缓冲液中甘油与溴酚蓝的左右分别是
① 上样缓冲液中包括甘油,溴酚蓝,SDS等,甘油主要作用是防止样品漂浮出点样孔,溴酚蓝作为电泳指示剂,用来观察电泳进度,SDS用于一些核酸结合蛋白的变性解离.
②核酸染料与凝胶中核酸结合,以便于在紫外光下显示核酸条带.
sds电泳上样缓冲液如何配置