OCT4 Recombinant Rabbit Monoclonal Antibody (3H8L6)

货号：701756

规格：100 µg

价格：4809

产品类型：流式抗体

品牌：Thermo Fisher

抗原：OCT4

物种：人/小鼠

宿主：兔

抗体亚型：IgG

克隆号：3H8L6

荧光染料：其它

类型:	单抗	同型对照：	IgG
免疫原性：	Protein corresponding to Human POU5F1 (aa 1-360)	用法：	2.5 µg x 10^6 cells(Chip);1.0-2.0 µg/mL(ICC);1.0-2.0 µg/mL(IF);1.0-2.0 µg/mL(WB)

Product Specific InformationThis antibody is predicted to react with Monkey, Goat, Cat and HorseRecombinant rabbit monoclonal antibodies are produced using in vitro expression systems. The expression systems are developed by cloning in the specific antibody DNA sequences from immunoreactive rabbits. Then, individual clones are screened to select the best candidates for production. The advantages of using recombinant rabbit monoclonal antibodies include: better specificity and sensitivity, lot-to-lot consistency, animal origin-free formulations, and broader immunoreactivity to diverse targets due to larger rabbit immune repertoire.Target InformationOct4 is a maternally expressed octamer-binding protein that was the first transcription factor described for the early stages of development. Oct4 is one of the four Yamaka factors, necessary for embryonic stem cell pluripotency. Pluripotent stem (iPS) cells, can be created by expressing Oct4 and the transcription factors Sox2, Klf4 and Lin28 along with c-Myc in mouse fibroblasts. More recently, experiments have demonstrated that iPS cells could be generated using expression plasmids expressing Oct4, Sox2, KlfF4 and c-Myc, eliminating the need for virus introduction, thereby addressing a safety concern for potential use of iPS cells in regenerative medicine. A translocation of this gene with the Ewing's sarcoma gene, t(6;22)(p21;q12), has been linked to tumor formation.For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

数据

OCT4 Antibody (701756) in IFFor immunofluorescence analysis, NTERA-2 cells were fixed and permeabilized for detection of endogenous OCT4 using ABfinity™ Anti- OCT4 Recombinant Rabbit Monoclonal Antibody (Product # 701756, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of OCT4 protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating nuclear localization of OCT4. Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification. OCT4 Antibody (701756) in WBWestern blot analysis was performed on whole cell extracts (30 µg lysate) of NTERA-2 (Lane 1), NCCIT (Lane 2), F9 (Lane 3), HEK-293 (Lane 4), A431 (Lane 5) and HeLa (Lane 6). The blots were probed with ABfinity™ Anti-OCT4 rabbit Monoclonal Antibody (Product # 701756, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 38 kDa band corresponding to OCT4 was observed only in specific cell lines. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), 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® 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). OCT4 Antibody (701756) in ChIPEnrichment of endogenous OCT4 protein at specific gene loci using ABfinity™ Anti-OCT4 Recombinant Rabbit Monoclonal Antibody: Chromatin Immunoprecipiation (ChIP) was performed using ABfinity™ Anti-OCT4 Recombinant Rabbit Monoclonal Antibody (Product # 701756, 5 µg) on sheared chromatin from 2 million NTERA2 cells using the "MAGnify ChIP system" kit (Product # 49-2024). Normal Rabbit IgG (1 µg) was used as a negative IP control. The purified DNA was analyzed by 7500 Fast qPCR system (Product # 4351106) with optimized PCR primer pairs for the promoters of the active SOX4, FOXA2, TNC region used as positive control target genes, and the region of the inactive MYOD, SAT2 satellite repeat, used as negative control target gene. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

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