Polyclonal Antibody to Erythropoietin (EPO)
EP; Epoetin; Erythropoetin; Hematopoietin; Hemopoietin
- Product No.PAA028Ra01
- Organism SpeciesRattus norvegicus (Rat) Same name, Different species.
- All
- Human
- Mouse
- Rat
- Cavia
- Rabbit
- Simian
- Caprine
- Ovine
- Equine
- Bovine
- Porcine
- Gallus
- Canine
- Others
- Multi-species
- Pan-species
- SourcePolyclonal antibody preparation
- HostRabbit
- Potencyn/a
- Ig TypeIgG
- PurificationAntigen-specific affinity chromatography followed by Protein A affinity chromatography
- LabelNone
- ImmunogenRPA028Ra01-Recombinant Erythropoietin (EPO)
- Buffer FormulationPBS, pH7.4, containing 0.02% NaN3, 50% glycerol.
- TraitsLiquid
- Concentration500µg/mL
- Organism Species Moren/a
- ApplicationsWB; IHC; ICC; IP.If the antibody is used in flow cytometry, please check FCM antibodies.
- DownloadInstruction Manual
- UOM20µl100µl200µl1ml10ml
- FOBUS$ 103 For more details, please contact local distributors!US$ 239 For more details, please contact local distributors!US$ 342 For more details, please contact local distributors!US$ 855 For more details, please contact local distributors!US$ 3420 For more details, please contact local distributors!
SPECIFITY of the Polyclonal Antibody to Erythropoietin (EPO)
The antibody isa rabbit polyclonal antibody raised against EPO. It has been selected for its ability to recognize EPO in immunohistochemical staining and western blotting.
USAGE of the Polyclonal Antibody to Erythropoietin (EPO)
Western blotting: 0.5-2ug/mlImmunocytochemistry in formalin fixed cells: 5-20ug/mlImmunohistochemistry in formalin fixed frozen section: 5-20ug/mlImmunohistochemistry in paraffin section: 5-20ug/mlEnzyme-linked Immunosorbent Assay: 0.05-2ug/mlOptimal working dilutions must be determined by end user.
STORAGE of the of the Polyclonal Antibody to Erythropoietin (EPO)
Store at 4°C for frequent use. Stored at -20°C in a manual defrost freezer for two year without detectable loss of activity. Avoid repeated freeze-thaw cycles.
STABILITY of the Polyclonal Antibody to Erythropoietin (EPO)
The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.
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INCREMENT SERVICES
Antibody Labeling Customized ServiceProtein A/G Purification ColumnStaining Solution for Cells and TissuePositive Controlfor AntibodyTissue/Sections Customized ServicePhosphorylated Antibody Customized ServiceWestern Blot (WB) Experiment ServiceImmunohistochemistry (IHC) Experiment ServiceImmunocytochemistry (ICC) Experiment ServiceFlow Cytometry (FCM) Experiment ServiceImmunoprecipitation (IP) Experiment ServiceImmunofluorescence (IF) Experiment ServiceBufferDAB Chromogen KitSABC KitLong-arm Biotin Labeling KitReal Time PCR Experimental Service
Related products
Catalog No. | Organism species: Rattus norvegicus (Rat) | Applications (RESEARCH USE ONLY!) |
APA028Ra61 | Active Erythropoietin (EPO) | Cell culture; Activity Assays. |
EPA028Ra61 | Eukaryotic Erythropoietin (EPO) | Positive Control; Immunogen; SDS-PAGE; WB. |
RPA028Ra01 | Recombinant Erythropoietin (EPO) | Positive Control; Immunogen; SDS-PAGE; WB. |
PAA028Ra01 | Polyclonal Antibody to Erythropoietin (EPO) | WB; IHC; ICC; IP. |
MAA028Ra21 | Monoclonal Antibody to Erythropoietin (EPO) | WB; IHC; ICC; IP. |
SEA028Ra | ELISA Kit for Erythropoietin (EPO) | Enzyme-linked immunosorbent assay for Antigen Detection. |
SCA028Ra | CLIA Kit for Erythropoietin (EPO) | Chemiluminescent immunoassay for Antigen Detection. |
KSA028Ra01 | ELISA Kit DIY Materials for Erythropoietin (EPO) | Main materials for "Do It (ELISA Kit) Yourself". |
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虽然没有具体用过检测这个抗原的抗体,但是如果希望抗体特异性好点,能检测丰度低的蛋白,还是选Abcam吧,Santa有时候全凭人品了,内参抗体什么的还能凑活,其他的比较勉强。
我觉得封闭这一步时间长短多少并不能减少背景…背景过深多半还是抗体浓度过大…不管一抗还是二抗过大都会使背景加深…一抗的浓度不一定是条带清晰就是最佳了…联系进一步优化一下
这个主要还是抗体的问题,WB最主要的还是要选择一个特异性较好的抗体,抗体是整个实验的重中之重
各位大神,有谁做WB,选用球虫的GADPH作为内参?知道哪个公司有针对球虫内参的抗体?
所以免疫组化在组织定位和定性上比较准确,但是在定量上不够准确。
western blot也是利用抗体和抗原之间的结合具有高度的特异性。经过聚丙烯酰胺凝胶电泳分离的蛋白质样品,转移到固相载体上,固相载体以非共价键形式吸附蛋白质,且能保持电泳分离的多肽类型及其生物学活性不变。以固相载体上的蛋白质或多肽作为抗原,与对应的抗体起免疫反应,再与酶或同位素标记的第二抗体起反应。
western blot因为有内参标定,所以在定量上要更加准确。但是在组织定位上不如免疫组化
一.样本种属来源:
首先要考虑的就是实验样本来源于什么物种。
1、哺乳动物的组织或者细胞样本,通常选择β-actin、β-tubulin、GAPDH、Lamin B、Histone H3、Na,K atpase等。
2、植物来源实验样本,则可以选择plantactin、Rubisco等。
3、其他来源样本研究较少,所以就应该参照文献报导,选择合适的蛋白作为内参。
二.目的蛋白分子量:
选择内参抗体时,应该考虑目的蛋白分子量的大小。通常应该保证目的蛋白与内参蛋白分子量相差5KD以上。比如目的蛋白分子量为45KD,此时不适宜选择β-actin作为内参,可以考虑选择GAPDH或者β-tubulin作为内参。
三.目的蛋白表达部位:
就一般的蛋白检测来说,β-actin、β-Tubulin抗体等就可以了,而针对于核蛋白的定量,特别是样本蛋白就是核蛋白时,选择恰当的核蛋白内参则更能体现内部参照的价值。常用的核内参抗体有Lamin A、Lamin B、Histone H3,除此之外,其它常见的核蛋白内参还有PCNA、K70、K80等,在一些文献报道中,Erk2、TATA binding protein(TBP)以及c-Jun、c-Fos等都有使用。而对于膜蛋白检测,常用的内参抗体为Na,K ATPase。对于线粒体蛋白的检测,常用VDAC1和COX IV作为内参抗体。
以上几条原则只是针对通常情况,但是需要注意的问题是——内参的选择需要考虑实际的试验环境,比如某些细胞中,由于组织缺氧、糖尿病等因素会导致GAPDH的表达增高,不适合做内参。比如在涉及细胞增殖相关试验中,c-Jun由于自身表达变化就不适合做内参;而在凋亡实验时,TBP、Lamin等也不适合作为内参。因此设计实验方案的时候应该考虑这些因素并查询相应文献,在实验过程中也应该注意如果内参表达出现异常,应考虑这方面因素。