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LFNG Recombinant Rabbit Monoclonal Antibody [JE66-62]

Usd: 385

Specification

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Catalog# HA721367

LFNG Recombinant Rabbit Monoclonal Antibody [JE66-62]

Application

  • WB

  • IF-Cell

  • FC

Reactivity

  • Human

  • Mouse

Conjugation

  • unconjugated

Overview

Product Name

LFNG Recombinant Rabbit Monoclonal Antibody [JE66-62]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human LFNG aa 330-379 / 379.

Species Reactivity

Human, Mouse

Validated Applications

WB, IF-Cell, FC

Molecular Weight

Predicted band size: 42 kDa

Positive Control

Hela cell lysate, K-562 cell lysate, HL-60 cell lysate, MCF7 cell lysate, HT-29 cell lysate, C2C12 cell lysate, PC-12 cell lysate, mouse testis tissue lysate, HUVEC, MCF7, K-562.

Conjugation

unconjugated

Clone Number

JE66-62

RRID

AB_3072484

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IF-Cell

  • 1:500

  • FC

  • 1:1,000

Target

Function

Glycosyltransferase that initiates the elongation of O-linked fucose residues attached to EGF-like repeats in the extracellular domain of Notch molecules. Modulates NOTCH1 activity by modifying O-fucose residues at specific EGF-like domains resulting in inhibition of NOTCH1 activation by JAG1 and enhancement of NOTCH1 activation by DLL1 via an increase in its binding to DLL1 (By similarity).

Background References

1. K Shimizu et al. Manic fringe and lunatic fringe modify different sites of the Notch2 extracellular region, resulting in different signaling modulation. J Biol Chem 276(28):25753-8 (2001)

2. Riley MF et al. Mir-125a-5p-mediated regulation of Lfng is essential for the avian segmentation clock. Dev Cell 24(5) (2013)

Subcellular Location

Golgi apparatus membrane.

UNIPROT

SwissProt: Q8NES3 Human

SwissProt: O09010 Mouse

Synonyms

Beta-1,3-N-acetylglucosaminyltransferase lunatic fringe

O-fucosylpeptide 3-beta-N-acetylglucosaminyltransferase

Images

  • Western blot analysis of LFNG on different lysates with Rabbit anti-LFNG antibody (<a href="/products/HA721367" style="font-weight: bold;text-decoration: underline;">HA721367</a>) at 1/1,000 dilution.<br /><br />Lane 1: Hela cell lysate (15 µg/Lane)<br />Lane 2: K-562 cell lysate (15 µg/Lane)<br />Lane 3: HL-60 cell lysate (15 µg/Lane)<br />Lane 4: MCF7 cell lysate (15 µg/Lane)<br />Lane 5: HT-29 cell lysate (15 µg/Lane)<br />Lane 6: C2C12 cell lysate (15 µg/Lane)<br />Lane 7: PC-12 cell lysate (15 µg/Lane)<br />Lane 8: Mouse testis tissue lysate (30 µg/Lane)<br /><br />Predicted band size: 42 kDa<br />Observed band size: 42 kDa<br /><br />Exposure time: 7 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721367" style="font-weight: bold;text-decoration: underline;">HA721367</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:100,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of LFNG on different lysates with Rabbit anti-LFNG antibody (HA721367) at 1/1,000 dilution.

    Lane 1: Hela cell lysate (15 µg/Lane)
    Lane 2: K-562 cell lysate (15 µg/Lane)
    Lane 3: HL-60 cell lysate (15 µg/Lane)
    Lane 4: MCF7 cell lysate (15 µg/Lane)
    Lane 5: HT-29 cell lysate (15 µg/Lane)
    Lane 6: C2C12 cell lysate (15 µg/Lane)
    Lane 7: PC-12 cell lysate (15 µg/Lane)
    Lane 8: Mouse testis tissue lysate (30 µg/Lane)

    Predicted band size: 42 kDa
    Observed band size: 42 kDa

    Exposure time: 7 seconds;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721367) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of HUVEC cells labeling LFNG with Rabbit anti-LFNG antibody (<a href="/products/HA721367" style="font-weight: bold;text-decoration: underline;">HA721367</a>) at 1/500 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-LFNG antibody (<a href="/products/HA721367" style="font-weight: bold;text-decoration: underline;">HA721367</a>) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HUVEC cells labeling LFNG with Rabbit anti-LFNG antibody (HA721367) at 1/500 dilution.

    Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-LFNG antibody (HA721367) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunocytochemistry analysis of MCF7 cells labeling LFNG with Rabbit anti-LFNG antibody (<a href="/products/HA721367" style="font-weight: bold;text-decoration: underline;">HA721367</a>) at 1/500 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-LFNG antibody (<a href="/products/HA721367" style="font-weight: bold;text-decoration: underline;">HA721367</a>) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of MCF7 cells labeling LFNG with Rabbit anti-LFNG antibody (HA721367) at 1/500 dilution.

    Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-LFNG antibody (HA721367) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Flow cytometric analysis of K-562 cells labeling LFNG.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA721367" style="font-weight: bold;text-decoration: underline;">HA721367</a>, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of K-562 cells labeling LFNG.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA721367, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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