Anti-Nanog Antibody Picoband™

Product Name

Anti-Nanog Antibody Picoband™

View all Nanog Antibodies

SKU/Catalog Number

A00153-4

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-Nanog Antibody Picoband™ catalog # A00153-4. Tested in ELISA, Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Mouse, Rat.

Storage & Handling

At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.

Cite This Product

Anti-Nanog Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00153-4)

Host

Rabbit

Contents

Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived mouse Nanog recombinant protein (Position: M1-A191).

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

A00153-4 is reactive to Nanog in Mouse, Rat

Applications

A00153-4 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

37 kDa

Calculated molecular weight

34.24kDa

Background of Nanog

NANOG (pron. nanOg) is a transcription factor critically involved with self-renewal of undifferentiated embryonic stem cells. In humans, this protein is encoded by the NANOG gene. It is mapped to 12p13.31. NANOG is thought to be a key factor in maintaining pluripotency. Moreover, NANOG is also thought to function in concert with other factors such as POU5F1 (Oct-4) and SOX2 to establish ESC identity. The NANOG protein has been found to be a transcriptional activator for the Rex1 promoter, playing a key role in sustaining Rex1 expression. Knockdown of NANOG in embryonic stem cells results in a reduction of Rex1 expression, while forced expression of NANOG stimulates Rex1 expression.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.25-0.5 μg/ml, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Rat
Flow Cytometry, 1-3 μg/1x10⁶ cells, Mouse
Direct ELISA, 0.1-0.5 μg/ml, Mouse

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of Nanog using anti-Nanog antibody (A00153-4).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: rat testis tissue lysates,
Lane 2: rat C6 whole cell lysates,
Lane 3: rat PC-12 whole cell lysates,
Lane 4: mouse testis tissue lysates,
Lane 5: mouse RAW264.7 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Nanog antigen affinity purified polyclonal antibody (Catalog # A00153-4) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Nanog at approximately 37 kDa. The expected band size for Nanog is at 37 kDa.

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Figure 2. IF analysis of Nanog using anti-Nanog antibody (A00153-4).
Nanog was detected in an immunocytochemical section of NRK cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-Nanog Antibody (A00153-4) overnight at 4°C. DyLight®594 Conjugated Goat Anti-Rabbit IgG (BA1142) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

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Figure 3. Flow Cytometry analysis of RAW264.7 cells using anti-Nanog antibody (A00153-4).
Overlay histogram showing RAW264.7 cells stained with A00153-4 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Nanog Antibody (A00153-4, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Gene/Protein Information For Nanog (Source: Uniprot.org, NCBI)

Alternative Names

FLJ12581; FLJ40451; hNanog; homeobox protein NANOG; Homeobox transcription factor Nanog; homeobox transcription factor Nanog-delta 48; Nanog homeobox; Nanog Nanog|2410002E02Rik, EN, ENK, ecat, ecat4|Nanog homeobox|homeobox protein NANOG|ES cell-associated protein 4|early embryo specific expression NK family|early embryo specific expression NK-type homeobox protein|homeobox transcription factor Nanog

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In this infographic, you will see the following information for Nanog: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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