![StressMarq/Anti-HO-1 Antibody [1F12-A6]/SMC-131D-APC/100-µg](images/StressMarq/201710/SMC-131_HO-1_Antibody_1F12-A6_ICC-IF_Human_HeLa-Cells_100x_Composite-150x150.png)
Overview:
| Product Name | HO-1 Antibody | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Description | Mouse Anti-Human HO-1 Monoclonal IgG1 Kappa | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Species Reactivity | Dog, Human, Monkey, Mouse, Rat, Bovine, Guinea Pig (Cavia porcellus), Hamster, Pig, Rabbit | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Applications | WB, IHC, ICC/IF, IP, ELISA | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Antibody Dilution | WB (1:1000), IHC (1:100), ICC/IF (1:100); optimal dilutions for assays should be determined by the user. | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Host Species | Mouse | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Immunogen Species | Human | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Immunogen | Human HO-1 synthetic peptide, amino acids 1-30 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 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
StreptavidinProperties:
Streptavidin Datasheet Biotin | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| R-PE (R-Phycoerythrin) | ||
Overview:
R-PE Datasheet |  | Optical Properties: λex = 565 nm λem = 575 nm εmax = 2.0×106 Φf = 0.84 Brightness = 1.68 x 103 Laser = 488 to 561 nm Filter set = TRITC |
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 | 1F12-A6 |
| Isotype | IgG1 Kappa |
| Specificity | Detects 32kDa. Does not cross-react with HO-2. |
| Cite This Product | StressMarq Biosciences Cat# SMC-131, RRID: AB_2264116 |
| Certificate of Analysis | 1 µg/ml was sufficient for detection of HO-1 in 10 µg of mixed human cell line lysate by colorimetric immunoblot analysis using Goat Anti-Mouse IgG:HRP as the secondary. |
Biological Description
| Alternative Names | HSP32 Antibody, HMOX1 Antibody, Heme oxygenase 1 Antibody, HO Antibody, HO1 Antibody |
| Research Areas | Cancer, Oxidative Stress |
| Cellular Localization | Endoplasmic Reticulum, Microsome |
| Accession Number | NP_002124.1 |
| Gene ID | 3162 |
| Swiss Prot | P09601 |
| Scientific Background | Heme-oxygenase is a ubiquitous enzyme that catalyzes the initial and rate-limiting steps in heme catabolism yielding equimolar amounts of biliverdin, iron and carbon monoxide. Biliverdin is subsequently converted to bilirubin and the free iron is sequestered to ferritin (1). These products have important physiological effects as carbon monoxide is a potent vasodilator; biliverdin and bilirubin are potent antioxidants; and the free iron increases oxidative stress and regulates the expression of many mRNAs (2). There are three isoforms of heme-oxygenase, HO-1, HO-2 and HO-3; however HO-1 and HO-2 are the major isoforms as they both have been identified in mammals (3). HO-1, also known as heat shock protein 32, is an inducible isoform activated by most oxidative stress inducers, cytokines, inflammatory agents and heat shock. HO-2 is a constitutive isoform which is expressed under homeostatic conditions. HO-1 is also considered to be a cytoprotective factor in that free heme is highly reactive and cytotoxic, and secondly, carbon monoxide is a mediator inhibiting the inflammatory process and bilirubin is a scavenger for reactive oxygen, both of which are the end products of heme catalyzation (4). It has also been shown that HO-1 deficiency may cause reduced stress defense, a pro-inflammatory tendency (5), susceptibility to atherosclerotic lesion formation (6), endothelial cell injury, and growth retardation (7). Up-regulation of HO-1 is therefore said to be one of the major defense mechanisms of oxidative stress (4). |
| References |
1. Froh M. et al. (2007) World J. Gastroentereol 13(25): 3478-86. 2. Elbirt K.K. and Bonkovsky H.L. (1999) Proc Assoc Am Physicians 111(5): 348-47. 3. Maines M.D., Trakshel G.M., and Kutty R.K. (1986) J Biol Chem 261: 411–419. 4. Brydun A., et al. (2007) Hypertens Res 30(4): 341-8. 5. Poss K.D. and Tonegawa S. (1997). Proc Natl Acad Sci U S A. 94: 10925–10930. 6. Yet S.F., et al. (2003) FASEB J. 17: 1759–1761. 7. Yachie A., et al. (1999) J Clin Invest. 103: 129–135. |
Product Images
Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-HO-1 Monoclonal Antibody, Clone 1F12-A6 (SMC-131). Tissue: HeLa Cells. Species: Human. Fixation: 2% Formaldehyde for 20 min at RT. Primary Antibody: Mouse Anti-HO-1 Monoclonal Antibody (SMC-131) at 1:100 for 12 hours at 4°C. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:200 for 2 hours at RT. Counterstain: DAPI (blue) nuclear stain at 1:40000 for 2 hours at RT. Localization: Microsome. Endoplasmic reticulum. Localizaes to the nucleus upon hypoxia. Magnification: 100x. (A) DAPI (blue) nuclear stain. (B) Anti-HO-1 Antibody. (C) Composite.
Immunohistochemistry analysis using Mouse Anti-HO-1 Monoclonal Antibody, Clone 1F12-A6 (SMC-131). Tissue: backskin. Species: Mouse. Fixation: Bouin’s Fixative and paraffin-embedded. Primary Antibody: Mouse Anti-HO-1 Monoclonal Antibody (SMC-131) at 1:100 for 1 hour at RT. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:50 for 1 hour at RT. Localization: muscle, dermis, hair follicles, epidermis: nuclear everywhere and some cytoplasmic staining.
Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-HO-1 Monoclonal Antibody, Clone 1F12-A6 (SMC-131). Tissue: HeLa Cells. Species: Human. Fixation: 2% Formaldehyde for 20 min at RT. Primary Antibody: Mouse Anti-HO-1 Monoclonal Antibody (SMC-131) at 1:100 for 12 hours at 4°C. Secondary Antibody: R-PE Goat Anti-Mouse (yellow) at 1:200 for 2 hours at RT. Counterstain: DAPI (blue) nuclear stain at 1:40000 for 2 hours at RT. Localization: Microsome. Endoplasmic reticulum. Localizaes to the nucleus upon hypoxia. Magnification: 20x. (A) DAPI (blue) nuclear stain. (B) Anti-HO-1 Antibody. (C) Composite.
Western Blot analysis of Human HeLa cell lysates showing detection of HO-1 protein using Mouse Anti-HO-1 Monoclonal Antibody, Clone 1F12-A6 (SMC-131). Load: 15 µg protein. Block: 1.5% BSA for 30 minutes at RT. Primary Antibody: Mouse Anti-HO-1 Monoclonal Antibody (SMC-131) at 1:1000 for 2 hours at RT. Secondary Antibody: Sheep Anti-Mouse IgG: HRP for 1 hour at RT.
Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-HO-1 Monoclonal Antibody, Clone 1F12-A6 (SMC-131). Tissue: HaCaT cells. Species: Human. Fixation: Cold 100% methanol for 10 minutes at -20°C. Primary Antibody: Mouse Anti-HO-1 Monoclonal Antibody (SMC-131) at 1:100 for 1 hour at RT. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:50 for 1 hour at RT. Localization: Cellcell border staining in epidermis, punctuate nuclear staining. .
Product Citations (4)
Western Blot
Nrf2 Activation Attenuates Both Orthodontic Tooth Movement and Relapse.
Kanzaki, H. et al. (2015) J Dent Res. 94(6):787-94.
PubMed ID: 25795629 Reactivity Mouse Applications: Western Blot
Characterization of the Anti-inflammatory Activity of Enones Based on the Evaluation of Their Heme Oxygenase-1 and Inducible NO Synthase Activity.
Rucker, H. et al. (2015) Universität Regensburg. PhD Dissertation
PubMed ID: N/A Reactivity Mouse Applications: Western Blot
Nuclear Nrf2 Induction by Protein Transduction Attenuates Osteoclastogenesis.
Kanzaki, H. et al. (2014) Free Radic Biol Med. 77:239-48.
PubMed ID: 25224039 Reactivity Mouse Applications: Western Blot
Opening or Closing the Lock? When Reactivity Is the Key to Biological Activity.
Al-Rifai, N., Rücker, H., Amslinger, S. (2013) Chemistry - A Euro J. 19(45):15384-95.
PubMed ID: 24105896 Reactivity Mouse Applications: Western Blot
| APC (Allophycocyanin) | ||
Overview:
APC Datasheet |  | Optical Properties: λex = 650 nm λem = 660 nm εmax = 7.0×105 Φf = 0.68 Brightness = 476 Laser = 594 or 633 nm Filter set = Cy®5 |
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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?
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！
两个CEX方法A和B测定同一单抗，结果碱性峰比例差不多，酸性峰比例相差约7%，相应主峰也差了7%左右。
具体来说，A方法酸性峰高，主峰低，碱性峰稍微低点；B方法酸性峰低，主峰高，碱性峰稍微高点；另外也做了CIEF，结果呢和A方法更接近。
仔细比较起来，AB两个方法的峰性和数量差不多，就不知道为什么会有这么大的差异。两个方法一个用的WCX柱-磷酸缓冲液，一个用SCX柱-MES缓冲液
大家帮我分析下：
1.两个方法哪个方法更准确，是以酸性峰高的为准还是什么？为什么？
2.这显著差异是由方法造成，具体原因是什么？柱子？
3.CIEF的结果和A方法更接近，是不是可以由此证明A方法更好或者CIEF的方法更好（因为CIEF更快更方便）？
欢迎讨论~
纠正下，A方法用的是Tosoh的柱子，B方法用的是SCX柱。TOSOH的柱子是7um的填料，10cm长。SCX是10um的填料。我本人TOSOH的阳离子柱子用的很少，这次信手用用，结果发现差异很大
那我现在就考虑，在以后方法开发过程中，除了通过流动相pH和组成、梯度、柱子选择来获得样品主峰和酸碱性的最大分离，还要关注各峰比例。因为之前比较方法好坏都只看分离度，尤其是主峰和邻近峰的分离度，获得最大分离度，自然可以做到主峰尽可能纯，但从未认真比较过各峰比例。这是一个大疏忽吧！
另外，CIEF和CEX方法原理还是有点差异的，所以分的是不同的异质体，原液放行两个方法肯定是都要做的。问题就是在早期细胞株筛选和工艺开发阶段，哪个方法才是又快又准。CIEF（iCE280)一般15分钟一个样，比CEX快多了。如果CIEF测得主峰要低于CEX结果，是不是真的完全可以取代CEX呢？CEX分离出的峰远比CIEF的多！
欢迎大家继续讨论~
因为是考察不同PH对药物的影响，样品又不好改变其PH值，这种情况怎么办？希望有经验的高手指教。
我的流动相是甲醇-水（90：10）
谢谢赐教！
请进子版按格式发贴，自行修改，谢谢。
这就是说不用酸碱预处理吗？
Whatman的网站上没有DE52最大耐受压力，请问又经验的战友应该是多少？
Whatman的网站上：
DE32DryMicrogranularDEAECellulose
SimilarperformancecharacteristicsafterprecyclingasDE52.
DE52PreswollenMicrogranularDEAECellulose
ProbablythemostwidelyusedDEAEcelluloseintheworld;usedforbiopolymerswithlowtohighnegativecharges;exhibitsexcellentresolutionwithgoodflowrates.
附件是一本图书(MethodsinMolecularMedicine,)的章节,上面说:
WhatmanDEAE52comesalreadypreswollenandonlyneedstobetransferred
totherunningbuffer50mMTE8.
lAntibodiesUsingIonExchangeChromatography.pdf(87.06k)
