AF1638

antibody from R&D Systems

Targeting: GNL3 C77032, E2IG3, MGC800, NS

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

AF1638

Provider

R&D Systems

Product name

Human/Mouse/Rat Nucleostemin Antibody

Antibody type

Polyclonal

Description

Antigen Affinity-purified. Detects human, mouse, rat Nucleostemin in direct ELISAs and Western blots. In direct ELISAs, less than 1% cross-reactivity with recombinant rat Nucleostemin C-terminal peptide (aa 281-538) is observed. Rat Nucleostemin specific IgG was purified by first passing the sera over a rat Nucleostemin aa 2-538 column and then passing the bound fraction over a rat Nucleostemin aa 281-538 column to removed rat Nucleostemin C-terminal specific IgG.

Reactivity

Human, Mouse, Rat

Host

Goat

Conjugate

Unconjugated

Antigen sequence

Q811S9


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

Phenotypic plasticity in normal breast derived epithelial cells.

Sauder CA, Koziel JE, Choi M, Fox MJ, Grimes BR, Badve S, Blosser RJ, Radovich M, Lam CC, Vaughan MB, Herbert BS, Clare SE
BMC cell biology 2014 Jun 10;15:20

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Overexpression of nucleostemin contributes to an advanced malignant phenotype and a poor prognosis in oral squamous cell carcinoma.

Yoshida R, Nakayama H, Nagata M, Hirosue A, Tanaka T, Kawahara K, Nakagawa Y, Matsuoka Y, Sakata J, Arita H, Hiraki A, Shinohara M, Ito T
British journal of cancer 2014 Dec 9;111(12):2308-15

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Nucleostemin is indispensable for the maintenance and genetic stability of hematopoietic stem cells.

Yamashita M, Nitta E, Nagamatsu G, Ikushima YM, Hosokawa K, Arai F, Suda T
Biochemical and biophysical research communications 2013 Nov 8;441(1):196-201

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Reactive oxygen species regulate nucleostemin oligomerization and protein degradation.

Huang M, Whang P, Chodaparambil JV, Pollyea DA, Kusler B, Xu L, Felsher DW, Mitchell BS
The Journal of biological chemistry 2011 Apr 1;286(13):11035-46

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Depletion of guanine nucleotides leads to the Mdm2-dependent proteasomal degradation of nucleostemin.

Huang M, Itahana K, Zhang Y, Mitchell BS
Cancer research 2009 Apr 1;69(7):3004-12

Western blot

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

Detection of Human Nucleostemin by Western Blot. Western blot shows lysates of PC-3 human prostate cancer cell line and SW480 human colorectal adenocarcinoma cell line. PVDF membrane was probed with 0.25 µg/mL of Goat Anti-Human/Mouse/Rat Nucleostemin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1638) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for Nucleostemin at approximately 70 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 Nucleostemin by Simple Western^TM. Simple Western lane view shows lysates of PC-3 human prostate cancer cell line, loaded at 0.2 mg/mL. A specific band was detected for Nucleostemin at approximately 83 kDa (as indicated) using 5 µg/mL of Goat Anti-Human/Mouse/Rat Nucleostemin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1638) 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

Nucleostemin in U2OS Human Cell Line. Nucleostemin was detected in immersion fixed U2OS human osteosarcoma cell line using 10 µg/mL Goat Anti-Human/Mouse/Rat Nucleostemin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1638) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red, upper panel; Catalog # NL001) and counterstained with DAPI (blue, lower panel). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.