Cyclophilin B Polyclonal Antibody

货号： PA1-027A

规格：100 µg

价格：4050

产品类型：流式抗体

品牌：Thermo Fisher

物种：人/小鼠/大鼠

宿主：兔

抗体亚型：IgG

荧光染料：其它

类型:	多抗	同型对照：	IgG
免疫原性：	Synthetic peptide corresponding to residues C(194) G K I E V E K P F A I A K E(208) of human CyPB	用法：	3-5 µg/1x10^6 cells(Flow);1:100-1:1000(IF);1:100-1:1000(ICC);1-2µg/mL(WB)

Product Specific InformationPA1-027A detects cyclophilin B (CyPB) in mouse, rat, canine, and human tissues.PA1-027A has been successfully used in Western blot procedures, IF/ICC and IHC (P). By Western blot, this antibody detects a ~19 kDa protein representing CyPB from rat liver extract.The PA1-027A immunogen is a synthetic peptide corresponding to residues C(194) G K I E V E K P F A I A K E(208) of human CyPB. This sequence is almost completely conserved between human, mouse, rat, bovine, and chicken. PA1-027A immunizing peptide (Cat. # PEP-058) is available for use in neutralization and control experiments.Target InformationImmunophilins are a family of soluble cytosolic receptors capable of binding to one of two major immunosuppressant agents - cyclosporin A (CsA) or FK506. Proteins which bind FK506 are termed FK506 Binding Proteins (FKBP's) and those which bind cyclosporin A are called cyclophilins (CyP).Both CyP:CsA and FKBP:FK506 complexes have been shown to inhibit calcineurin, a calcium and calmodulin dependent protein phosphatase which has been implicated as an important signaling enzyme in T-cell activation. Thus, providing a possible mechanism of immunosuppression by CsA and FK506. Immunophilins function as peptidyl prolyl cis-trans-isomerases (PPIase) whose activity is inhibited by their respective immunosuppressant compounds. As PPIase's, immunophilins accelerate folding of some proteins both in vivo and in vitro by catalyzing slow steps in the initial folding and rearrangement of proline containing proteins.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

数据

Cyclophilin B Antibody (PA1-027A) in IFImmunofluorescent analysis of Cyclophilin B (green) showing staining in the in the cytoplasm of A431 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Cyclophilin B polyclonal antibody (Product # PA1-027A) in 3% BSA-PBS at a dilution of 1:500 and incubated overnight at 4°C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x. Cyclophilin B Antibody (PA1-027A) in IHC (P)Immunohistochemistry analysis of Cyclophilin B showing staining in the cytoplasm of paraffin-embedded human hepatocarcinoma (right) compared with a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Cyclophilin B polyclonal antibody (Product # PA1-027A) diluted in 3% BSA-PBS at a dilution of 1:200 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting. Cyclophilin B Antibody (PA1-027A) in WBWestern blot analysis was performed on whole cell extracts (30 µg lysate) of U-87 MG (Lane 1), IMR-32 (Lane 2), PC-3 (Lane 3), U-2 OS (Lane 4), SH-SY5Y (Lane 5), Jurkat (Lane 6), and MCF 7 (Lane 7). The blots were probed with Anti-Cyclophilin B Rabbit Polyclonal Antibody (Product # PA1-027A, 1-2 µg/mL) and detected by chemiluminescence using Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (Product # G-21234, 1:5000 dilution). A ~ 23 kDa band corresponding to Cyclophilin B was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0302BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

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参考文献：

1.Molecular and cellular endocrinologyExpression of Peroxisome Proliferator-Activated Receptor alpha (PPARα) in somatotropinomas: Relationship with Aryl hydrocarbon receptor Interacting Protein (AIP) and in vitro effects of fenofibrate in GH3 cells."PA1-027A was used in western blot to determine the relationship with aryl hydrocarbon receptor interacting protein (AIP) and in vitor effects of fenofibrate in GH3 cells to study expression of peroxisome proliferator-activated receptor alpha (PPAR-alpha)"AuthorsRotondi S,Modarelli A,Oliva MA,Rostomyan L,Sanita P,Ventura L,Daly AF,Esposito V,Angelucci A,Arcella A,Giangaspero F,Beckers A,Jaffrain-Rea ML2.American journal of physiology. Endocrinology and metabolismThyroid hormone and estradiol have overlapping effects on kidney glutathione S-transferase-α gene expression."PA1-027A was used in western blot to compare the effects of thyroid hormone and estradiol on kidney GST-? expression"AuthorsFaustino LC,Almeida NA,Pereira GF,Ramos RG,Soares RM,Morales MM,Pazos-Moura CC,Ortiga-Carvalho TM3.Molecular pharmaceuticsSmall interfering RNA knocks down the molecular target of alendronate, farnesyl pyrophosphate synthase, in osteoclast and osteoblast cultures."PA1-027A was used in western blot to evaluate a small interfering RNA against farnesyl pyrophosphate synthase"AuthorsWang Y,Panasiuk A,Grainger DW4.The Journal of endocrinologyThyroid hormone contributes to the hypolipidemic effect of polyunsaturated fatty acids from fish oil: in vivo evidence for cross talking mechanisms."PA1-027A was used in western blot to investigate the role of thyroid hormone signaling in the hypolipidemic effect of n-3 polyunsaturated fatty acidsn-3"AuthorsSouza LL,Cordeiro A,Oliveira LS,de Paula GS,Faustino LC,Ortiga-Carvalho TM,Oliveira KJ,Pazos-Moura CC5.Nature cell biologyAnalysis of the myosin-II-responsive focal adhesion proteome reveals a role for β-Pix in negative regulation of focal adhesion maturation."PA1-027A was used in western blot to study the role of myosin II activity in focal adhesion using proteomics"AuthorsKuo JC,Han X,Hsiao CT,Yates JR,Waterman CM

技术参数

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