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ARF5 Recombinant Rabbit Monoclonal Antibody [JE53-83]

Usd: 385

Specification

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Catalog# ET7110-46

ARF5 Recombinant Rabbit Monoclonal Antibody [JE53-83]

Application

  • WB

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

ARF5 Recombinant Rabbit Monoclonal Antibody [JE53-83]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human ARF5 aa 60-180 / 180.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P

Molecular Weight

Predicted band size: 21 kDa

Positive Control

HeLa cell lysate, HepG2 cell lysate, A549 cell lysate, PC-12 cell lysate, mouse brain tissue lysate, human liver tissue, human breast carcinoma tissue, mouse lung tissue.

Conjugation

unconjugated

Clone Number

JE53-83

RRID

AB_3071011

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. 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

  • 1:2,000

  • IHC-P

  • 1:50-1:200

Target

Function

The ADP-ribosylation factor (ARF) protein family are structurally and functionally conserved members of the Ras superfamily of regulatory GTP-binding proteins. ARFs influence vesicle trafficking and signal transduction in eukaryotic cells. ARF-dependent regulatory mechanisms include the coordination of spectrin interactions with golgi membranes and the association of actin to the golgi via rho family-dependent G-protein localization (Rac, CDC42) and WASP/Arp2/3 complexes. Additionally, ARFs play a central role in maintenance of organelle integrity, assembly of coat proteins, and activation of phospho-lipase D. The ARF proteins are categorized as class I (ARF1, ARF2, and ARF3), class II (ARF4 and ARF5) and class III (ARF6); members of each class share a common gene organization.

Background References

1. Liu Z. et. al. ARF2-ARF4 and ARF5 are Essential for Female and Male Gametophyte Development in Arabidopsis. Plant Cell Physiol. 2018 Jan 1;59(1):179-189.

2. Egerer J. et. al. GORAB Missense Mutations Disrupt RAB6 and ARF5 Binding and Golgi Targeting. J Invest Dermatol. 2015 Oct;135(10):2368-2376.

Sequence Similarity

Belongs to the small GTPase superfamily. Arf family.

Subcellular Location

Golgi apparatus, trans-Golgi network membrane, perinuclear region, membrane.

UNIPROT

SwissProt: P84085 Human

SwissProt: P84084 Mouse

SwissProt: P84083 Rat

Synonyms

ADP ribosylation factor 5 antibody

ADP-ribosylation factor 5 antibody

ARF 5 antibody

Arf5 antibody

ARF5_HUMAN antibody

Images

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of ARF5 on different lysates with Rabbit anti-ARF5 antibody (<a href="/products/ET7110-46" style="font-weight: bold;text-decoration: underline;">ET7110-46</a>) at 1/1,000 dilution.<br /><br />Lane 1: A549-WT cell lysate<br />Lane 2: A549-KD ARF5 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 21 kDa<br />Observed band size: 18 kDa<br /><br />Exposure time: 2 minutes 30 seconds; ECL: K1802;<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/ET7110-46" style="font-weight: bold;text-decoration: underline;">ET7110-46</a>) at 1/1,000 dilution was used in 5% BSA at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of ARF5 on different lysates with Rabbit anti-ARF5 antibody (ET7110-46) at 1/1,000 dilution.

    Lane 1: A549-WT cell lysate
    Lane 2: A549-KD ARF5 cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 21 kDa
    Observed band size: 18 kDa

    Exposure time: 2 minutes 30 seconds; ECL: K1802;

    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 (ET7110-46) at 1/1,000 dilution was used in 5% BSA at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of ARF5 on different lysates with Rabbit anti-ARF5 antibody (<a href="/products/ET7110-46" style="font-weight: bold;text-decoration: underline;">ET7110-46</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa cell lysate (20 µg/Lane)<br />Lane 2: HepG2 cell lysate (20 µg/Lane)<br />Lane 3: A549 cell lysate (20 µg/Lane)<br />Lane 4: PC-12 cell lysate (20 µg/Lane)<br />Lane 5: Mouse brain tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 21 kDa<br />Observed band size: 18 kDa<br /><br />Exposure time: 3 minutes; ECL: K1801;<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/ET7110-46" style="font-weight: bold;text-decoration: underline;">ET7110-46</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of ARF5 on different lysates with Rabbit anti-ARF5 antibody (ET7110-46) at 1/2,000 dilution.

    Lane 1: HeLa cell lysate (20 µg/Lane)
    Lane 2: HepG2 cell lysate (20 µg/Lane)
    Lane 3: A549 cell lysate (20 µg/Lane)
    Lane 4: PC-12 cell lysate (20 µg/Lane)
    Lane 5: Mouse brain tissue lysate (40 µg/Lane)

    Predicted band size: 21 kDa
    Observed band size: 18 kDa

    Exposure time: 3 minutes; ECL: K1801;

    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 (ET7110-46) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-ARF5 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7110-46" style="font-weight: bold;text-decoration: underline;">ET7110-46</a>, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-ARF5 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7110-46, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-ARF5 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7110-46" style="font-weight: bold;text-decoration: underline;">ET7110-46</a>, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-ARF5 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7110-46, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse lung tissue using anti-ARF5 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7110-46" style="font-weight: bold;text-decoration: underline;">ET7110-46</a>, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse lung tissue using anti-ARF5 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7110-46, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

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