Stem Cell Markers: Antibodies to Induced Pluripotent Stem Cells - FocusOn 055

Introduction

Induced pluripotent stem cells (iPS cells or iPSCs) are produced from non-pluripotent cells, via programmed expression of specific genes via human intervention. The extent of match between natural pluripotent stem cells and those derived artificially via gene manipulation is still under determination, but congruence exists for a number of signatures such as morphology, chromatin methylation patterns, stem cell markers, telomerase activity, doubling rates, teratoma formation, and embryoid body formation.

Advantages of iPSCs are the possibility to avoid immune rejection by patients since they are produced from the patients’ own adult somatic cells, and to circumvent the controversial use of embryos in stem cell research. Disadvantages include the propensity of iPSCs to form tumors.

The genes demonstrated to date to promote iPSC formation have all been associated with cancer by one means or another. Alternative methods to produce iPSCs, including non-genetic methods, as well as to increase efficiency of iPSC formation, are under active research to address significant barriers to safe clinical application in regenerative medicine.

iPS cell generation is crucially dependent on the role of specific genes, with high interested for targets identified by current research as follows:

Oct3/4

Oct3/4, also known as POU5F1, is a transcription factor essential to maintaining pluripotency. The presence of Oct3/4 permits pluripotency and differentiation potential of embryonic stem cells, while its absence blocks such potential.

Sox family

The Sox family of transcription factors is also associated with maintaining pluripotency. Sox1 and Sox2 yield iPS cells with similar efficiency. Sox3, Sox15, and Sox18 also generate iPS cells, although with relative decreased efficiency.

Fig. 1: Formalin-fixed paraffin-embedded (FFPE) human prostate stained with Oct3/4 antibody Cat.-No. AP07188PU-N
Fig. 1: Formalin-fixed paraffin-embedded (FFPE) human prostate stained with Oct3/4 antibody Cat.-No. AP07188PU-N
Fig. 2: Human skin (FFPE) stained with anti-Sox2 antibody Cat.-No. AP15787PU-S
Fig. 2: Human skin (FFPE) stained with anti-Sox2 antibody Cat.-No. AP15787PU-S

Klf family

The Klf (Krueppel-like factor) family of transcription factors have been scrutinized extensively for their roles in cell proliferation, differentiation and survival, particularly with respect to cancer. Klf4 was initially identified for its role in the artificial generation of mouse iPS cells – further research has not proven the necessity of Klf genes in the generation of human iPS cells. Klf2 shows similar efficiency in iPS production, while family members Klf1 and Klf5 produce iPS with greatly reduced efficiency.

Fig. 3: Immunofluorescence analysis of Sox2 monoclonal antibody Cat.-No. AM11050PU-N in HeLa cells at 25µg/ml followed by Alexa-Fluor-546-conjugated secondary antibody (orange; blue counterstaining with DAPI)
Fig. 3: Immunofluorescence analysis of Sox2 monoclonal antibody Cat.-No. AM11050PU-N in HeLa cells at 25µg/ml followed by Alexa-Fluor-546-conjugated secondary antibody (orange; blue counterstaining with DAPI)
Fig. 4: Colon (FFPE) stained with Klf4 antibody Cat.-No. AP08292PU-N
Fig. 4: Colon (FFPE) stained with Klf4 antibody Cat.-No. AP08292PU-N

Myc family

The Myc family of genes (c-Myc, n-Myc, and l-Myc) are protooncogenes strongly tied to cancer. C-Myc is a factor identified in the generation of mouse iPS cells; like Klf family members, research is not proven the requirement for Myc genes in the generation of human iPS cells. A quarter of mice transplanted with c-Myc-induced iPS cells develop lethal teratomas, which has spurred research into methods to avoid the use of Myc genes in production of induced pluripotent stem cells.

Nanog

In embryonic stem cells, Nanog is necessary, along with Oct-3/4 and Sox2, to promote pluripotency. Research has shown it is possible to manufacture iPS cells with Nanog as a contributing factor, while other research has observed that Nanog is unnecessary for induction.

Lin28

Lin28 is an mRNA binding protein expressed in embryonic stem cells and embryonic carcinoma cells associated with differentiation and proliferation. It has been demonstrated as a factor in iPSC generation, although its presence is not necessary.

Further readings

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Primary Antibodies

Catalog No. Host Iso. Clone Pres. React. Applications  

c-Myc antibody

c-Myc Mouse IgG1 33 Purified Av, Hu, Ms, Rt C, IP, WB
0.1 mg / €360.00
  Acris Antibodies GmbH

c-Myc antibody

Western blot analysis of extracts from NIH/3T3 cells treated with EGF using Myc antibody. Rabbit Aff - Purified Hu, Ms, Rt WB
0.1 ml / €270.00
  Acris Antibodies GmbH

c-Myc antibody

Western blot analysis of extracts from NIH/3T3 cells treated with EGF using Myc antibody. Rabbit Aff - Purified Hu, Ms, Rt WB
50 µl / €230.00
  Acris Antibodies GmbH

KLF4 (1-170) antibody

Immunohistochemistry: Paraffin embedded sections of human gastric cancer tissue were incubated with anti-human KLF4 (1:50) for 2 hours at room temperature. Antigen retrieval was performed in 0.1 M sodium citrate buffer. Diaminobenzidine (DAB) was used for detection. Mouse IgG1 AT4E6 Purified Hu E, F, P, WB
0.1 ml / €310.00
  Acris Antibodies GmbH

KLF4 (1-170) antibody

Immunohistochemistry: Paraffin embedded sections of human gastric cancer tissue were incubated with anti-human KLF4 (1:50) for 2 hours at room temperature. Antigen retrieval was performed in 0.1 M sodium citrate buffer. Diaminobenzidine (DAB) was used for detection. Mouse IgG1 AT4E6 Purified Hu E, F, P, WB
50 µl / €230.00
  Acris Antibodies GmbH

KLF4 antibody

Figure 2. Immunoflurescence analysis of KLF4 antibody AM11052PU-N (5 ug/ml). Hela cells transfected with pMX constructs of human KLF4 (top) and NIH3T3 cells transfected with pMX constructs of mouse KLF4 (bottom) were analyzed at approximately 62 hours after transfection. Mouse IgG1 56CT5.1.6 Purified Hu ICC/IF, WB
0.4 ml / €370.00
  Acris Antibodies GmbH

KLF4 Control Peptide

KLF4
50 µg / €160.00
  Acris Antibodies GmbH

LIN28 antibody

Figure 2. Formalin-fixed and paraffin-embedded human testis tissue reacted with LIN28A Monoclonal Antibody (Cat.#AM11048PU-N), which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry. Clinical relevance has not been evaluated. Mouse IgG1 55CT58.12.1 Purified Hu P, WB

Regular Price: 0.4 ml / €370.00

Special Price: 0.4 ml / €231.00

  Acris Antibodies GmbH

LIN28 Control Peptide

LIN28
50 µg / €160.00
  Acris Antibodies GmbH

NANOG (1-154) antibody

Immunohistochemistry: Paraffin Embedded Sections of Human seminoma tissue were incubated with anti-Human Nanog (1/50) for 2 hours at RT. Antigen retrieval was performed in 0.1M sodium citrate buffer and detected using Diaminobenzidine (DAB). Mouse IgG2a 5A10 Purified Hu E, P, WB
0.1 ml / €310.00
  Acris Antibodies GmbH

NANOG antibody

Immunohistochemistry: Paraffin Embedded Sections of Human seminoma tissue were incubated with anti-Human Nanog (1/50) for 2 hours at RT. Antigen retrieval was performed in 0.1M sodium citrate buffer and detected using Diaminobenzidine (DAB). Mouse IgG2a 5A10 Purified Hu E, P, WB
50 µl / €230.00
  Acris Antibodies GmbH

NANOG antibody

Immunohistochemistry analysis in formalin fixed and paraffin embedded human testis tissue using anti-NANOG monoclonal antibody Cat.-No AM11049SU-N followed by peroxidase conjugation of the secondary antibody and DAB staining.This data demonstrates the use of NANOG Antibody for immunohistochemistry. Clinical relevance has not been evaluated. Mouse IgG1 60CT70.5.12 Purified Hu P, WB
0.4 ml / €370.00
  Acris Antibodies GmbH

OCT3/4 antibody

AP15694PU Oct3/Oct4 antibody staining of Human Seminoma. Rabbit Purified Hu P
1 ml / €520.00
  Acris Antibodies GmbH

OCT3/4 antibody

AP15694PU Oct3/Oct4 antibody staining of Human Seminoma. Rabbit Purified Hu P
0.1 ml / €190.00
  Acris Antibodies GmbH

OCT3/4 antibody

Formalin fixed paraffin embedded Testicular Seminoma stained with OCT4 antibody Cat.-No AM03173PU (Clone NRG1.1) Mouse IgG2b NRG1.1 Purified Hu P
1 ml / €610.00
  Acris Antibodies GmbH

OCT3/4 antibody

Formalin fixed paraffin embedded Testicular Seminoma stained with OCT4 antibody Cat.-No AM03173PU (Clone NRG1.1) Mouse IgG2b NRG1.1 Purified Hu P
0.5 ml / €410.00
  Acris Antibodies GmbH

OCT3/4 Control Peptide

OCT3/4 Purified
50 µg / €190.00
  Acris Antibodies GmbH

OCT3/4 antibody

Western blot analysis of Oct-3/4 in rat kidney tissue lysate using AP00249PU-N. Rabbit Aff - Purified Bov, Hu, Ms, Por, Rt IP, P, WB
0.1 mg / €460.00
  Acris Antibodies GmbH

OCT3/4 antibody

WB using Anti-Oct4 antibody shows detection of endogenous Oct4 in mouse embryonic fibroblast (MEF) cell lysate (lane 1) and HeLa nuclear extract (HN) (lane 2). The band at ~39 kDa (arrow head) corresponds to Oct4. After transfer, the membrane was blocked with 5% BSA. Primary antibody was used at a 1:1,000 dilution in PBS containing 5% BSA. The specificity of the antibody was confirmed by peptide competition which blocks reaction of the antibody with Oct4 (lanes 3 and 4, respectively). Rabbit IgG Aff - Purified Chimp, Eq, Hu, Macaque, Ms E, WB
25 µl / €190.00
  Acris Antibodies GmbH

OCT3/4 (N-term) antibody

Immunofluorescence analysis of OCT3 (OCT4) antibody (N-term) (AP11910PU-N) in HeLa cells. 0.025 mg/ml primary antibody was followed by Alexa-Fluor-546-conjugated donkey anti-rabbit lgG (H+L). Alexa-Fluor-546 emits orange fluorescence. Blue counterstaining is DAPI. Rabbit Ig Purified Hu ICC/IF, WB
0.4 ml / €370.00
  Acris Antibodies GmbH

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