Anti-Nucleolin Monoclonal Antibody (4E2)

Product Name

Anti-Nucleolin Monoclonal Antibody (4E2)

View all Nucleolin Antibodies

SKU/Catalog Number

M00228-1

Size

100µg

Form

Liquid. In PBS containing 50% glycerol.

Description

Boster Bio Anti-Nucleolin Monoclonal Antibody (4E2) catalog # M00228-1. Tested in IF, IHC, WB applications. This antibody reacts with Human.

Storage & Handling

Store at -20°C for one year. For short term storage and frequent use, store at 4°C for up to one month. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-Nucleolin Monoclonal Antibody (4E2) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M00228-1)

Host

Mouse

Contents

Liquid. In PBS containing 50% glycerol.

Clonality

Monoclonal

Clone Number

4E2

Isotype

IgG1, kappa

Immunogen

Human nucleolin from Raji cell extract.

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

Reactive Species

M00228-1 is reactive to NCL in Human

Applications

M00228-1 is guaranteed for IF, IHC, WB Boster Guarantee

Observed Molecular Weight

Calculated molecular weight

76.614kDa

Background of Nucleolin

Recognizes a protein of 70kDa, which is identified as CD86 (HLDA V; WS Code BP BP072. HLDA V; WS Code A A109. HLDA VI; WS Code BP 95. HLDA VI; WS Code B CD86.9). CD86 is a type I transmembrane glycoprotein and a member of the immunoglobulin superfamily of cell surface receptors. It is expressed at high levels on resting peripheral monocytes and dendritic cells and at very low density on resting B and T lymphocytes. CD86 expression is rapidly upregulated by B cell specific stimuli with peak expression at 18 to 42 hours after stimulation. CD86, along with CD80/B71, is an important accessory molecule in T cell co-stimulation via its interaction with CD28 and CD152/CTLA4. Since CD86 has rapid kinetics of induction, it is believed to be the major CD28 ligand expressed early in the immune response. It is also found on malignant Hodgkin and Reed Sternberg (HRS) cells in Hodgkin's disease.

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

Restore with deionized water (or equivalent) for reconstitution volume of 100 µL

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.

Flow Cytometry (5-10µg/ml final concentration)
Immunocytochemistry (1-10µg/ml)
Immunohistochemistry (1-10µg/ml)
Western Blot (1µg/ml, ECL)
Suggested dilutions/conditions may not be available for all applications.
Optimal conditions must be determined individually for each application.

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of NCL using anti-NCL antibody (M00228-1).
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 50ug of sample under reducing conditions.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-NCL antigen affinity purified polyclonal antibody (Catalog # M00228-1) at 0.5 ug/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-Mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # SA1021) with Tanon 5200 system. A specific band was detected for NCL.

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Figure 2. IHC analysis of NCL using anti-NCL antibody (M00228-1).
NCL was detected in paraffin-embedded section. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-NCL Antibody (M00228-1) overnight at 4°C. Biotinylated goat anti Mouse IgG antibody was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.

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

Alternative Names

C23; FLJ45706; FLJ59041; nucleolin; Protein C23 NCL C23, Nsr1 nucleolin nucleolin

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