Overview:
Product Name | Smad3 Antibody | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Description | Rabbit Anti-Human Smad3 Polyclonal | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Species Reactivity | Human, Mouse, Rat | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Applications | WB, IHC | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Antibody Dilution | WB (1:2000), IHC (1:200); optimal dilutions for assays should be determined by the user. | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Host Species | Rabbit | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Immunogen Species | Human | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Immunogen | A synthesized peptide derived from human Smad3 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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
StreptavidinProperties:
Streptavidin Datasheet BiotinProperties:
Biotin Datasheet HRP (Horseradish peroxidase)Properties:
HRP Datasheet AP (Alkaline Phosphatase)Properties:
AP Datasheet
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Properties
Storage Buffer | PBS pH 7.4, 50% glycerol, 150mM NaCl, 0.02% sodium azide |
Storage Temperature | -20ºC |
Shipping Temperature | Blue Ice or 4ºC |
Purification | Affinity Purified |
Clonality | Polyclonal |
Isotype | IgG |
Specificity | Detects ~48kDa, endogenous levels of total Smad3 |
Cite This Product | StressMarq Biosciences Cat# SPC-1285, RRID: AB_2712935 |
Certificate of Analysis | A 1:1000 dilution of SPC-1285 was sufficient for detection of Smad3 in 10 µg of Hek293 cell lysates by ECL immunoblot analysis using Goat Anti-Rabbit IgG:HRP as the secondary antibody. |
Biological Description
Alternative Names | hMAD 3 Antibody, JV15 2 Antibody, LDS1C Antibody, MAD (mothers against decapentaplegic Drosophila) homolog 3 Antibody, MAD homolog 3 Antibody, Mad3 Antibody, MADH3 Antibody, SMAD 3 Antibody |
Research Areas | Cell Signaling, Cytoplasmic, Epigenetics and Nuclear Signaling, Epigenetics and Nuclear Signalling, Nuclear Signaling Pathways, Signaling Pathways, SMADs, Stem Cells, TGF Beta |
Cellular Localization | Cytoplasm, Nucleus |
Accession Number | NP_005893.1 |
Gene ID | 4088 |
Swiss Prot | P84022 |
Scientific Background | Members of the Smad family of cell signaling molecules are components of a critical intracellular pathway that transmit TGF-β signals from the cell surface into the nucleus. There are three distinct classes of Smads: the receptor-regulated Smads (R-Smads), the common-mediator Smad (co-Smad), and the antagonistic or inhibitory Smads. Following stimulation by TGF-β, Smad2 and Smad3 become phosphorylated at their carboxyl termini (Ser465 and 467 on Smad2; Ser423 and 425 on Smad3) by TGF-β Receptor I. Phosphorylated Smad 2/3 can complex with Smad4, translocate to the nucleus and regulate gene expression. |
References |
1. Heldin C.H. et al. (1997) Nature. 390: 465-471. 2. Attisano L. and Wrana, J.L. (1998) Curr Opin Cell Biol. 10: 188-194. 3. Massagué J. (1998) Annu Rev Biochem. 67: 753-791. 4. Whitman M. (1998) Genes Dev. 12: 2445-2462. 5. Wu G. et al. (2000) Science. 287: 92-97. 6. Abdollah S., et al. (1997) J. Biol. Chem. 272: 27678-27685. 7. Souchelnytskyi S., et al. (1997) J. Biol. Chem. 272: 28107-28115. 8. Liu X., et al. (1997) Proc. Natl. Acad. Sci. USA. 94: 10669-10674. |
Product Images
Western blot analysis of Human Hek293 cell lysates showing detection of ~58kDa Smad3 protein using Rabbit Anti-Smad3 Polyclonal Antibody (SPC-1285). Lane 1: Human Hek293 treated with the immunizing peptide. Lane 2: Human Hek293. Primary Antibody: Rabbit Anti-Smad3 Polyclonal Antibody (SPC-1285) at 1:1000. Predicted/Observed Size: ~58kDa.
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1.直接用固体磷酸钠配制成50mM的磷酸钠溶液,再调pH到7.4;(我们试着用这个做了下,发现挂不上柱)
2.配置磷酸钠盐缓冲液:按NaH2PO4:Na2HPO4以19:81的摩尔比配制成pH7.4的缓冲液?(附一张百度出来的配方
)
3.如果是磷酸钠盐缓冲液,可以直接将50mM的NaH2PO4的水溶液用NaOH调成pH7.4吗?
再者,2和3这两个方法配制的磷酸钠盐缓冲液有什么区别?最终效果是一样的吗?如果不一样,有什么理论的知识支撑呢?个人感觉是分析化学中酸碱理论中的缓冲液那里的知识。求帮忙解答这些疑问。
另外,我还想问一下,pH对于Ni柱对His-tagged的蛋白的分离纯化影响大吗?是怎么影响的?谢谢大家了!
由弱酸及其盐、弱碱及其盐组成的混合溶液,能在一定程度上抵消、减轻外加强酸或强碱对溶液酸碱度的影响,从而保持溶液的pH值相对稳定。这种溶液称为缓冲溶液。
是否可以理解为纯化水得PH范围为6.3-7.6?能否直接用pH计测量?谢谢!
pH(1)=pKa+lg[c(CH₃COONa)/c(CH₃COOH)]=pKa=4.74
通HCl后,溶液是c(CH₃COOH)=0.2mol/L、c(NaCl)=0.1mol/L的混合溶液,溶液pH按照弱酸溶液pH的求法求.
c(H⁺)=√[Ka*c(CH₃COOH)]=√(10^-4.74*0.2)=0.00191(mol/L)(采用了近似公式)
pH(2)=-lg{c(H⁺)}=2.72
两个pH求得,那么pH的变化量也就可得了.pH的变化量=|pH(2)-pH(1)|=|2.72-4.74|=2.02
1)PH缓冲溶液作用原理和pH值
当往某些溶液中加入一定量的酸和碱时,有阻碍溶液pH变化的作用,称为缓冲作用,这样的溶液叫做缓冲溶液.弱酸及其盐的混合溶液(如HAc与NaAc),弱碱及其盐的混合溶液(如NH3·H2O与NH4Cl)等都是缓冲溶液.
由弱酸HA及其盐NaA所组成的缓冲溶液对酸的缓冲作用,是由于溶液中存在足够量的碱A-的缘故.当向这种溶液中加入一定量的强酸时,H离子基本上被A-离子消耗:
所以溶液的pH值几乎不变;当加入一定量强碱时,溶液中存在的弱酸HA消耗OH-离子而阻碍pH的变化.
2)PH缓冲溶液的缓冲能力
在缓冲溶液中加入少量强酸或强碱,其溶液pH值变化不大,但若加入酸,碱的量多时,缓冲溶液就失去了它的缓冲作用.这说明它的缓冲能力是有一定限度的.
缓冲溶液的缓冲能力与组成缓冲溶液的组分浓度有关.0.1mol·L-1HAc和0.1mol·L-1NaAc组成的缓冲溶液,比0.01mol·L-1HAc和0.01mol·L-1NaAc的缓冲溶液缓冲能力大.关于这一点通过计算便可证实.但缓冲溶液组分的浓度不能太大,否则,不能忽视离子间的作用.
组成缓冲溶液的两组分的比值不为1∶1时,缓冲作用减小,缓冲能力降低,当c(盐)/c(酸)为1∶1时△pH最小,缓冲能力大.不论对于酸或碱都有较大的缓冲作用.缓冲溶液的pH值可用下式计算:
此时缓冲能力大.缓冲组分的比值离1∶1愈远,缓冲能力愈小,甚至不能起缓冲作用.对于任何缓冲体系,存在有效缓冲范围,这个范围大致在pKaφ(或pKbφ)两侧各一个pH单位之内.
弱酸及其盐(弱酸及其共轭碱)体系pH=pKaφ±1
弱碱及其盐(弱碱及其共轭酸)体系pOH=pKbφ±1
例如HAc的pKaφ为4.76,所以用HAc和NaAc适宜于配制pH为3.76~5.76的缓冲溶液,在这个范围内有较大的缓冲作用.配制pH=4.76的缓冲溶液时缓冲能力最大,此时(c(HAc)/c(NaAc)=1.
3)PH缓冲溶液的配制和应用
为了配制一定pH的缓冲溶液,首先选定一个弱酸,它的pKaφ尽可能接近所需配制的缓冲溶液的pH值,然后计算酸与碱的浓度比,根据此浓度比便可配制所需缓冲溶液.
以上主要以弱酸及其盐组成的缓冲溶液为例说明它的作用原理、pH计算和配制方法.对于弱碱及其盐组成的缓冲溶液可采用相同的方法.
PH缓冲溶液在物质分离和成分分析等方面应用广泛,如鉴定Mg2离子时,可用下面的反应:
白色磷酸铵镁沉淀溶于酸,故反应需在碱性溶液中进行,但碱性太强,可能生成白色Mg(OH)2沉淀,所以反应的pH值需控制在一定范围内,因此利用NH3·H2O和NH4Cl组成的缓冲溶液,保持溶液的pH值条件下,进行上述反应.
:)
我在做一细菌不同酸碱度生长状况时,发现这些奇怪现象:pH=3的培养基灭菌(TSB液体培养基)灭菌后pH上升到到9.2!而原来pH=9.0的降到8.7(基本没多少变化),请问各位大侠,这是什么原因?
一般做不同酸碱度生长实验时,该如何才能防止pH在湿热灭菌后基本不变化?
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