Product Details

Uniprot No.

P19338

Target Names

Alternative Names

C23 antibody; FLJ45706 antibody; MS1116 antibody; NCL antibody; Nucl antibody; NUCL_HUMAN antibody; Nucleolin antibody; Protein C23 antibody

Species Reactivity

Human, Rat

Immunogen

A synthesized peptide from human NCL protein

Immunogen Species

Homo sapiens (Human)

Conjugate

Non-conjugated

Clonality

Monoclonal

Isotype

Rabbit IgG

Clone No.

25D2

Purification Method

Affinity-chromatography

Concentration

It differs from different batches. Please contact us to confirm it.

Buffer

Rabbit IgG in 10mM phosphate buffered saline , pH 7.4, 150mM sodium chloride, 0.05% BSA, 0.02% sodium azide and 50% glycerol.

Form

Liquid

Tested Applications

ELISA, WB, IHC, IF, FC

Recommended Dilution

Application	Recommended Dilution
WB	1:500-1:5000
IHC	1:50-1:200
IF	1:50-1:200
FC	1:50-1:200

Protocols

ELISA Protocol
Western Blotting(WB) Protocol
Immunohistochemistry (IHC) Protocol
Immunofluorescence (IF) Protocol
Flow Cytometry (FC) Protocol

Troubleshooting and FAQs

Antibody FAQs

Storage

Upon receipt, store at -20°C or -80°C. Avoid repeated freeze.

Lead Time

Basically, we can dispatch the products out in 1-3 working days after receiving your orders. Delivery time maybe differs from different purchasing way or location, please kindly consult your local distributors for specific delivery time.

Description

Nucleolin is a multifunctional phosphoprotein predominantly localized in the nucleolus, where it plays essential roles in ribosome biogenesis, chromatin remodeling, and cell proliferation. Beyond its nuclear functions, nucleolin also shuttles to the cell surface in many cell types, making it a valuable marker for studying cellular growth dynamics and a target of interest in cancer research, where its expression is frequently upregulated.

This recombinant rabbit monoclonal antibody, clone 25D2, offers the reproducibility and consistency that demanding experimental workflows require. Generated against a synthetic peptide derived from human nucleolin and produced using recombinant technology, this antibody provides sequence-defined specificity and eliminates the lot-to-lot variability often encountered with traditional hybridoma-derived reagents. Affinity-purified to ensure high purity, it delivers reliable performance across multiple applications.

Validation studies demonstrate robust detection in western blot analysis across diverse human cell lines including HeLa, HEK293, A549, THP-1, and U-87MG, as well as rat brain tissue, confirming cross-species reactivity. The observed band at approximately 100 kDa, compared to the predicted 77 kDa molecular weight, reflects the extensive post-translational modifications characteristic of nucleolin, particularly phosphorylation and glycosylation. Immunohistochemistry validation in paraffin-embedded human liver cancer tissue shows clear staining, while immunofluorescence studies in HeLa cells reveal the expected nucleolar localization pattern. Flow cytometry analysis using Jurkat cells further demonstrates the antibody's versatility for quantitative single-cell applications.

Whether investigating ribosomal biology, examining proliferation markers, or exploring nucleolin's emerging roles in cancer cell biology, this antibody provides the multi-platform flexibility researchers need to advance their studies with confidence.

Usage

For Research Use Only. Not for use in diagnostic or therapeutic procedures.

Code	CSB-RA997430A0HU
Size	US$210
	Order now
Image	Western Blot Positive WB detected in: Hela whole cell lysate(30µg), HEK293 whole cell lysate(30µg), A549 whole cell lysate(30µg), THP-1 whole cell lysate(30µg), U-87MG whole cell lysate(30µg), Rat brain tissue lysate(30µg) All lanes: NCL antibody at 1:1000 Secondary Goat polyclonal to rabbit IgG at 1/40000 dilution Predicted band size: 77 kDa Observed band size: 100 kDa Exposure time：10s IHC image of CSB-RA997430A0HU diluted at 1:100 and staining in paraffin-embedded human liver cancer performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a Goat anti-rabbit polymer IgG labeled by HRP and visualized using 0.05% DAB. Immunofluorescence staining of HeLa cell with CSB-RA997430A0HU at 1:50 , counter-stained with DAPI. The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4°C. The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Rabbit IgG(H+L). Overlay Peak curve showing jurkat cells stained with CSB-RA997430A0HU (red line) at 1:100. The cells were fixed in 4% formaldehyde and permeated by 0.2% TritonX-100 for 10min. Then 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (1ug/110⁶cells) for 45min at 4℃. The secondary antibody used was FITC-conjugated goat anti-rabbit IgG (H+L) at 1/200 dilution for 35min at 4℃.Control antibody (green line) was Rabbit IgG (1ug/110⁶cells) used under the same conditions. Acquisition of >10, 000 events was performed.
Have Questions?	Leave a Message or Start an on-line Chat

NCL Recombinant Monoclonal Antibody

Product Details

Related Products

Customer Reviews and Q&A

Target Background