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Slit2 Recombinant Rabbit Monoclonal Antibody [JE31-87]

Usd: 385

Specification

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Catalog# HA721407

Slit2 Recombinant Rabbit Monoclonal Antibody [JE31-87]

Application

  • WB

  • IF-Cell

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

Slit2 Recombinant Rabbit Monoclonal Antibody [JE31-87]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human Slit2 aa 1,380-1,529 / 1,529.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, FC

Molecular Weight

Predicted band size: 170 kDa

Positive Control

HEK-293 cell lysate, 293T cell lysate, PC-3M cell lysate, Raji cell lysate, HCT 116 cell lysate, HeLa cell lysate, rat pancreas tissue lysate, SH-SY5Y cell lysate, human brain tissue lysate, mouse brain tissue lysate, rat brain tissue lysate, A549.

Conjugation

unconjugated

Clone Number

JE31-87

RRID

AB_3072524

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

  • 1:100

  • FC

  • 1:50-1:1,000

Target

Function

SLIT2 thought to act as molecular guidance cue in cellular migration, and function appears to be mediated by interaction with roundabout homolog receptors. During neural development involved in axonal navigation at the ventral midline of the neural tube and projection of axons to different regions. SLIT1 and SLIT2 seem to be essential for midline guidance in the forebrain by acting as repulsive signal preventing inappropriate midline crossing by axons projecting from the olfactory bulb. In spinal cord development may play a role in guiding commissural axons once they reached the floor plate by modulating the response to netrin. In vitro, silences the attractive effect of NTN1 but not its growth-stimulatory effect and silencing requires the formation of a ROBO1-DCC complex. May be implicated in spinal cord midline post-crossing axon repulsion. In vitro, only commissural axons that crossed the midline responded to SLIT2. In the developing visual system appears to function as repellent for retinal ganglion axons by providing a repulsion that directs these axons along their appropriate paths prior to, and after passage through, the optic chiasm. In vitro, collapses and repels retinal ganglion cell growth cones. Seems to play a role in branching and arborization of CNS sensory axons, and in neuronal cell migration. In vitro, Slit homolog 2 protein N-product, but not Slit homolog 2 protein C-product, repels olfactory bulb (OB) but not dorsal root ganglia (DRG) axons, induces OB growth cones collapse and induces branching of DRG axons. Seems to be involved in regulating leukocyte migration.

Background References

1. Geraldo LH et al. SLIT2/ROBO signaling in tumor-associated microglia and macrophages drives glioblastoma immunosuppression and vascular dysmorphia. J Clin Invest. 2021 Aug

2. Ahirwar DK et al. Slit2 Inhibits Breast Cancer Metastasis by Activating M1-Like Phagocytic and Antifibrotic Macrophages. Cancer Res. 2021 Oct

Subcellular Location

Secreted.

UNIPROT

SwissProt: O94813 Human

SwissProt: Q9R1B9 Mouse

SwissProt: Q9WVC1 Rat

Synonyms

Drad 1 antibody

E030015M03Rik antibody

E130320P19Rik antibody

FLJ14420 antibody

OTTHUMP00000158695 antibody

OTTHUMP00000217852 antibody

OTTHUMP00000217853 antibody

OTTHUMP00000217854 antibody

Slil 3 antibody

Slil3 antibody

Expand

Images

  • Western blot analysis of Slit2 on different lysates with Rabbit anti-Slit2 antibody (<a href="/products/HA721407" style="font-weight: bold;text-decoration: underline;">HA721407</a>) at 1/1,000 dilution.<br /><br />Lane 1: HEK-293 cell lysate (15 µg/Lane)<br />Lane 2: 293T cell lysate (15 µg/Lane)<br />Lane 3: PC-3M cell lysate (15 µg/Lane)<br />Lane 4: Raji cell lysate (15 µg/Lane)<br />Lane 5: HCT 116 cell lysate (15 µg/Lane)<br />Lane 6: HeLa cell lysate (15 µg/Lane)<br />Lane 7: Rat pancreas tissue lysate (30 µg/Lane)<br /><br />Predicted band size: 170 kDa<br />Observed band size: 50/200 kDa<br /><br />Exposure time: 3 minutes;<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/HA721407" style="font-weight: bold;text-decoration: underline;">HA721407</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 Slit2 on different lysates with Rabbit anti-Slit2 antibody (HA721407) at 1/1,000 dilution.

    Lane 1: HEK-293 cell lysate (15 µg/Lane)
    Lane 2: 293T cell lysate (15 µg/Lane)
    Lane 3: PC-3M cell lysate (15 µg/Lane)
    Lane 4: Raji cell lysate (15 µg/Lane)
    Lane 5: HCT 116 cell lysate (15 µg/Lane)
    Lane 6: HeLa cell lysate (15 µg/Lane)
    Lane 7: Rat pancreas tissue lysate (30 µg/Lane)

    Predicted band size: 170 kDa
    Observed band size: 50/200 kDa

    Exposure time: 3 minutes;

    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 (HA721407) 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.

  • Western blot analysis of Slit2 on different lysates with Rabbit anti-Slit2 antibody (<a href="/products/HA721407" style="font-weight: bold;text-decoration: underline;">HA721407</a>) at 1/1,000 dilution.<br /><br />Lane 1: SH-SY5Y cell lysate (15 µg/Lane)<br />Lane 2: Human brain tissue lysate (30 µg/Lane)<br />Lane 3: Mouse brain tissue lysate (30 µg/Lane)<br />Lane 4: Rat brain tissue lysate (30 µg/Lane)<br /><br />Predicted band size: 170 kDa<br />Observed band size: 50/200 kDa<br /><br />Exposure time: 28 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/HA721407" style="font-weight: bold;text-decoration: underline;">HA721407</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 Slit2 on different lysates with Rabbit anti-Slit2 antibody (HA721407) at 1/1,000 dilution.

    Lane 1: SH-SY5Y cell lysate (15 µg/Lane)
    Lane 2: Human brain tissue lysate (30 µg/Lane)
    Lane 3: Mouse brain tissue lysate (30 µg/Lane)
    Lane 4: Rat brain tissue lysate (30 µg/Lane)

    Predicted band size: 170 kDa
    Observed band size: 50/200 kDa

    Exposure time: 28 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 (HA721407) 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 A549 cells labeling Slit2 with Rabbit anti-Slit2 antibody (<a href="/products/HA721407" style="font-weight: bold;text-decoration: underline;">HA721407</a>) at 1/100 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-Slit2 antibody (<a href="/products/HA721407" style="font-weight: bold;text-decoration: underline;">HA721407</a>) at 1/100 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 A549 cells labeling Slit2 with Rabbit anti-Slit2 antibody (HA721407) at 1/100 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-Slit2 antibody (HA721407) at 1/100 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 A549 cells labeling Slit2.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA721407" style="font-weight: bold;text-decoration: underline;">HA721407</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 A549 cells labeling Slit2.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA721407, 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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