KLF4 Polyclonal Antibody

货号： PA1-095

规格：200 µL

价格：5467

产品类型：免疫组化一抗

品牌：Thermo Fisher

抗原：KLF4

物种：人/小鼠

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：	Recombinant N-terminal fragment (amino acids 1-118) of human KLF4	用法：	1-2.0 µg/mL(ICC);1-µg/mL(IF);1:500-1:2000(WB)

Product Specific InformationThe predicted molecular weight of KLF4 is ~55kD. The migration of this protein by SDS-PAGE was observed slightly higher (~60kD) with both PA1-095 and a benchmark antibody.PA1-095 detects a predominant band at ~60kD, and a nonspecific band of unknown origin at ~80kD in both whole cell lysates and nuclear extracts.For best results, StartingBlock T20 (TBS) Blocking Buffer (Product # 37543) is recommended.Target InformationKLF4 (Kruppel-like factor 4 (gut)) is a transcription factor that can act both as an activator and as a repressor. KLF4 binds to the 5'-CACCC-3' core sequence, and binds to the promoter region of its own gene and it can activate its own transcription. It regulates the expression of key transcription factors during embryonic development. KLF4 plays an important role in maintaining embryonic stem cells, and in preventing their differentiation. It is required for establishing the barrier function of the skin and for postnatal maturation and maintenance of the ocular surface. It is also involved in the differentiation of epithelial cells and may also function in skeletal and kidney development. Diseases associated with KLF4 include skin sequamous cell carcinoma and familial adenomatous polyposis.For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

数据

KLF4 Antibody (PA1-095) in IFImmunofluorescent analysis of KLF4 (red) in human embryonic stem cell H9 line grown on irradiated MEF-feeder layer. The cells were fixed with 4% paraformaldehyde at room temperature for 10 min and permeabilized with 0.25% Triton-X 100 for 5 min and blocked with the 10% BSA in PBS for 30 min at 37°C. Cells were stained with a KLF4 polyclonal antibody (Product # PA1-095) at a dilution of 1:200 in 3% BSA/PBS blocking buffer overnight at 4°C, and then incubated with a RRX-conjugated donkey anti-rabbit IgG secondary antibody at a dilution of 1:500 for 1 hour at room temperature. Nucleus DNA (blue) was stained with DAPI (Product # D1306). KLF4 Antibody (PA1-095) in IFImmunofluorescence analysis of KLF4 was done on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with KLF4 Rabbit Polyclonal Antibody (Product # PA1-095) at 1 µg/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing nuclear localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.KLF4 Antibody (PA1-095) in WBWestern blot analysis was performed on whole cell extracts (20 µg lysate) of HT-29 (Lane 1), HeLa (Lane 2), A-431 (Lane 3), PC-3 (lane 4), Jurkat (lane 5), HCT 116 (lane 6), K562 (lane 7), COLO 205 (lane 8), PC-12 (lane 9) and A549 (lane 10). The blots were probed with Anti-KLF4 Rabbit Polyclonal Antibody (Product # PA1-095, 1:500-1:1500 dilution) 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 54 kDa band corresponding to KLF4 was observed across cell lines tested expect for PC-12. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX), 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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