AF1759

antibody from R&D Systems

Targeting: POU5F1 MGC22487, OCT3, Oct4, OTF3

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Data presented on provider website)

Chromatin Immunoprecipitation (Supportive data in Antibodypedia)

Antibody Data

Product number

AF1759

Provider

R&D Systems

Product name

Human/Mouse Oct-3/4 Antibody

Antibody type

Polyclonal

Description

Antigen Affinity-purified. Detects human Oct-3/4 in direct ELISAs and Western blots.

Reactivity

Human, Mouse

Host

Goat

Conjugate

Unconjugated

Antigen sequence

Q01860


Isotype

IgG

Vial size

100 ug

Concentration

LYOPH

Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 12 months from date of receipt, -20 to -70 °C as supplied. 1 month, 2 to 8 °C under sterile conditions after reconstitution. 6 months, -20 to -70 °C under sterile conditions after reconstitution.

References

Submitted references

SOX2 and SOX9 are markers of clinically aggressive disease in metastatic high-grade serous carcinoma.

Sherman-Samis M, Onallah H, Holth A, Reich R, Davidson B
Gynecologic oncology 2019 Jun;153(3):651-660

---

The BAF and PRC2 Complex Subunits Dpf2 and Eed Antagonistically Converge on Tbx3 to Control ESC Differentiation.

Zhang W, Chronis C, Chen X, Zhang H, Spalinskas R, Pardo M, Chen L, Wu G, Zhu Z, Yu Y, Yu L, Choudhary J, Nichols J, Parast MM, Greber B, Sahlén P, Plath K
Cell stem cell 2019 Jan 3;24(1):138-152.e8

---

O-GlcNAc transferase activates stem-like cell potential in hepatocarcinoma through O-GlcNAcylation of eukaryotic initiation factor 4E.

Cao B, Duan M, Xing Y, Liu C, Yang F, Li Y, Yang T, Wei Y, Gao Q, Jiang J
Journal of cellular and molecular medicine 2019 Apr;23(4):2384-2398

---

Small non-coding RNA landscape of extracellular vesicles from human stem cells.

Kaur S, Abu-Shahba AG, Paananen RO, Hongisto H, Hiidenmaa H, Skottman H, Seppänen-Kaijansinkko R, Mannerström B
Scientific reports 2018 Oct 19;8(1):15503

---

Disrupted alternative splicing for genes implicated in splicing and ciliogenesis causes PRPF31 retinitis pigmentosa.

Buskin A, Zhu L, Chichagova V, Basu B, Mozaffari-Jovin S, Dolan D, Droop A, Collin J, Bronstein R, Mehrotra S, Farkas M, Hilgen G, White K, Pan KT, Treumann A, Hallam D, Bialas K, Chung G, Mellough C, Ding Y, Krasnogor N, Przyborski S, Zwolinski S, Al-Aama J, Alharthi S, Xu Y, Wheway G, Szymanska K, McKibbin M, Inglehearn CF, Elliott DJ, Lindsay S, Ali RR, Steel DH, Armstrong L, Sernagor E, Urlaub H, Pierce E, Lührmann R, Grellscheid SN, Johnson CA, Lako M
Nature communications 2018 Oct 12;9(1):4234

---

Generation of nine induced pluripotent stem cell lines as an ethnic diversity panel.

Gao X, Yourick JJ, Sprando RL
Stem cell research 2018 Aug;31:193-196

---

Dinaciclib potently suppresses MCL-1 and selectively induces the cell death in human iPS cells without affecting the viability of cardiac tissue.

Alsayegh K, Matsuura K, Sekine H, Shimizu T
Scientific reports 2017 Mar 31;7:45577

---

Generation of a TLE3 heterozygous knockout human embryonic stem cell line using CRISPR-Cas9.

Bara AM, Messana A, Herring A, Hazelbaker DZ, Eggan K, Barrett LE
Stem cell research 2016 Sep;17(2):441-443

---

Generation of a TLE1 homozygous knockout human embryonic stem cell line using CRISPR-Cas9.

Herring A, Messana A, Bara AM, Hazelbaker DZ, Eggan K, Barrett LE
Stem cell research 2016 Sep;17(2):430-432

---

Brain tissue banking for stem cells for our future.

Palmero E, Palmero S, Murrell W
Scientific reports 2016 Dec 19;6:39394

---

Semaphorin 3A induces mesenchymal-stem-like properties in human periodontal ligament cells.

Wada N, Maeda H, Hasegawa D, Gronthos S, Bartold PM, Menicanin D, Fujii S, Yoshida S, Tomokiyo A, Monnouchi S, Akamine A
Stem cells and development 2014 Sep 15;23(18):2225-36

---

Effects of antioxidants on the quality and genomic stability of induced pluripotent stem cells.

Luo L, Kawakatsu M, Guo CW, Urata Y, Huang WJ, Ali H, Doi H, Kitajima Y, Tanaka T, Goto S, Ono Y, Xin HB, Hamano K, Li TS
Scientific reports 2014 Jan 21;4:3779

---

Pluripotent stem cells derived from mouse and human white mature adipocytes.

Jumabay M, Abdmaulen R, Ly A, Cubberly MR, Shahmirian LJ, Heydarkhan-Hagvall S, Dumesic DA, Yao Y, Boström KI
Stem cells translational medicine 2014 Feb;3(2):161-71

---

Genome-wide chromatin interactions of the Nanog locus in pluripotency, differentiation, and reprogramming.

Apostolou E, Ferrari F, Walsh RM, Bar-Nur O, Stadtfeld M, Cheloufi S, Stuart HT, Polo JM, Ohsumi TK, Borowsky ML, Kharchenko PV, Park PJ, Hochedlinger K
Cell stem cell 2013 Jun 6;12(6):699-712

---

Generation of corneal epithelial cells from induced pluripotent stem cells derived from human dermal fibroblast and corneal limbal epithelium.

Hayashi R, Ishikawa Y, Ito M, Kageyama T, Takashiba K, Fujioka T, Tsujikawa M, Miyoshi H, Yamato M, Nakamura Y, Nishida K
PloS one 2012;7(9):e45435

---

A novel role for an RNA polymerase III subunit POLR3G in regulating pluripotency in human embryonic stem cells.

Wong RC, Pollan S, Fong H, Ibrahim A, Smith EL, Ho M, Laslett AL, Donovan PJ
Stem cells (Dayton, Ohio) 2011 Oct;29(10):1517-27

---

Glial cell line-derived neurotrophic factor and endothelial cells promote self-renewal of rabbit germ cells with spermatogonial stem cell properties.

Kubota H, Wu X, Goodyear SM, Avarbock MR, Brinster RL
FASEB journal : official publication of the Federation of American Societies for Experimental Biology 2011 Aug;25(8):2604-14

---

SIP1 mediates cell-fate decisions between neuroectoderm and mesendoderm in human pluripotent stem cells.

Chng Z, Teo A, Pedersen RA, Vallier L
Cell stem cell 2010 Jan 8;6(1):59-70

---

miR-302b maintains "stemness" of human embryonal carcinoma cells by post-transcriptional regulation of Cyclin D2 expression.

Lee NS, Kim JS, Cho WJ, Lee MR, Steiner R, Gompers A, Ling D, Zhang J, Strom P, Behlke M, Moon SH, Salvaterra PM, Jove R, Kim KS
Biochemical and biophysical research communications 2008 Dec 12;377(2):434-440

---

Regulation of self-renewal and pluripotency by Sox2 in human embryonic stem cells.

Fong H, Hohenstein KA, Donovan PJ
Stem cells (Dayton, Ohio) 2008 Aug;26(8):1931-8

---

Differential requirements for hematopoietic commitment between human and rhesus embryonic stem cells.

Rajesh D, Chinnasamy N, Mitalipov SM, Wolf DP, Slukvin I, Thomson JA, Shaaban AF
Stem cells (Dayton, Ohio) 2007 Feb;25(2):490-9

---

A comparison of NIH-approved human ESC lines.

Ware CB, Nelson AM, Blau CA
Stem cells (Dayton, Ohio) 2006 Dec;24(12):2677-84

---

Controlled-rate freezing of human ES cells.

Ware CB, Nelson AM, Blau CA
BioTechniques 2005 Jun;38(6):879-80, 882-3

Western blot

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

Detection of Human and Mouse Oct-3/4 by Western Blot. Western blot shows lysates of BG01V human embryonic stem cells and D3 mouse embryonic stem cell line. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human/Mouse Oct-3/4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1759) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Oct-3/4 at approximately 48 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

Detection of Human Oct-3/4 by Simple Western^TM. Simple Western lane view shows lysates of BG01V human embryonic stem cells and NTera-2 human testicular embryonic carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Oct-3/4 at approximately 50 kDa (as indicated) using 20 µg/mL of Goat Anti-Human/Mouse Oct-3/4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1759) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Immunocytochemistry

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

Alkaline Phosphatase and Oct-3/4 in BG01V Human Stem Cells. Alkaline phosphatase (ALPL) and Oct-3/4 were detected in human BG01V embryonic stem cells using 10 µg/mL Human/Mouse/Rat ALPL Monoclonal Antibody (Catalog # MAB1448) and 10 µg/mL Goat Anti-Human/Mouse Oct-3/4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1759). Cells were incubated with primary antibodies for 3 hours at room temperature. Cells were stained for ALPL using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (pseudo-stained green; Catalog # NL007), and stained for Oct-3/4 using the NorthernLights 637-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL002). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Chromatin Immunoprecipitation

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

Detection of Oct-3/4-regulated Genes by Chromatin Immunoprecipitation. BG01V human embryonic stem cells were fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. Oct-3/4/DNA complexes were immunoprecipitated using 5 μg Goat Anti-Human/Mouse Oct-3/4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1759) or control antibody (Catalog # AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (Catalog # BAF109). Immunocomplexes were captured using 50 μL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. The nanog promoter was detected by standard PCR.