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CCL5 Recombinant Rabbit Monoclonal Antibody [JM03-45]

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Specification

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Catalog# ET1705-70

CCL5 Recombinant Rabbit Monoclonal Antibody [JM03-45]

Application

  • WB

  • IHC-P

  • IF-Cell

  • IF-Tissue

Reactivity

  • Human

  • Mouse

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

CCL5 Recombinant Rabbit Monoclonal Antibody [JM03-45]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human RANTES aa 1-91.

Species Reactivity

Human, Mouse

Validated Applications

WB, IHC-P, IF-Cell, IF-Tissue

Molecular Weight

Predicted band size: 10 kDa

Positive Control

NK-92 cell lysates, mouse thymus lysates, NK-92, human tonsil tissue, human placenta tissue.

Conjugation

unconjugated

Clone Number

JM03-45

RRID

AB_3070600

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

  • 1:500-1:5,000

  • IF-Cell

  • 1:50-1:200

  • IF-Tissue

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

Target

Function

Chemoattractant for blood monocytes, memory T-helper cells and eosinophils. Causes the release of histamine from basophils and activates eosinophils. Binds to CCR1, CCR3, CCR4 and CCR5. One of the major HIV-suppressive factors produced by CD8+ T-cells. Recombinant RANTES protein induces a dose-dependent inhibition of different strains of HIV-1, HIV-2, and simian immunodeficiency virus (SIV). The processed form RANTES(3-68) acts as a natural chemotaxis inhibitor and is a more potent inhibitor of HIV-1-infection. The second processed form RANTES(4-68) exhibits reduced chemotactic and HIV-suppressive activity compared with RANTES(1-68) and RANTES(3-68) and is generated by an unidentified enzyme associated with monocytes and neutrophils.

Background References

1. Liu Y et al. TWEAK/Fn14 Activation Participates in Ro52-Mediated Photosensitization in Cutaneous Lupus Erythematosus. Front Immunol 8:651 (2017).

2. Fan Y et al. Low intraprostatic DHT promotes the infiltration of CD8+ T cells in BPH tissues via modulation of CCL5 secretion. Mediators Inflamm 2014:397815 (2014).

Sequence Similarity

Belongs to the intercrine beta (chemokine CC) family.

Tissue Specificity

Expressed in the follicular fluid (at protein level). T-cell and macrophage specific.

Post-translational Modification

N-terminal processed form RANTES(3-68) is produced by proteolytic cleavage, probably by DPP4, after secretion from peripheral blood leukocytes and cultured sarcoma cells.; The identity of the O-linked saccharides at Ser-27 and Ser-28 are not reported in PubMed:1380064. They are assigned by similarity.

Subcellular Location

Secreted.

UNIPROT

SwissProt: P13501 Human

SwissProt: P30882 Mouse

Synonyms

Beta chemokine RANTES antibody

Beta chemokine RANTES precursor antibody

C C motif chemokine 5 antibody

CCL 5 antibody

CCL5 antibody

CCL5_HUMAN antibody

Chemokine (C C motif) ligand 5 antibody

Chemokine CC Motif Ligand 5 antibody

D17S136E antibody

EoCP antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of CCL5 on different lysates with Rabbit anti-CCL5 antibody (<a href="/products/ET1705-70" style="font-weight: bold;text-decoration: underline;">ET1705-70</a>) at 1/5,000 dilution.<br /><br />Lane 1: NK-92 cell lysate (15 µg/Lane)<br />Lane 2: Raji cell lysate (negative) (15 µg/Lane)<br /><br />Predicted band size: 10 kDa<br />Observed band size: 10 kDa<br /><br />Exposure time: 15 seconds; 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/ET1705-70" style="font-weight: bold;text-decoration: underline;">ET1705-70</a>) at 1/5,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) 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.

    ☑ Relative expression (RE)

    Western blot analysis of CCL5 on different lysates with Rabbit anti-CCL5 antibody (ET1705-70) at 1/5,000 dilution.

    Lane 1: NK-92 cell lysate (15 µg/Lane)
    Lane 2: Raji cell lysate (negative) (15 µg/Lane)

    Predicted band size: 10 kDa
    Observed band size: 10 kDa

    Exposure time: 15 seconds; 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 (ET1705-70) at 1/5,000 dilution was used in primary antibody dilution (K1803) 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 CCL5 on mouse thymus lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (<a href="/products/ET1705-70" style="font-weight: bold;text-decoration: underline;">ET1705-70</a>, 1/500) was used in 5% BSA 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 1:40,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of CCL5 on mouse thymus lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1705-70, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:40,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of NK-92 cells labeling CCL5 with Rabbit anti-CCL5 antibody (<a href="/products/ET1705-70" style="font-weight: bold;text-decoration: underline;">ET1705-70</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-CCL5 antibody (<a href="/products/ET1705-70" style="font-weight: bold;text-decoration: underline;">ET1705-70</a>) at 1/100 dilution in 1% BSA in PBST 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. PBS instead of the primary antibody was used as the secondary antibody only control. 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 1/100 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 NK-92 cells labeling CCL5 with Rabbit anti-CCL5 antibody (ET1705-70) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-CCL5 antibody (ET1705-70) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-CCL5 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/ET1705-70" style="font-weight: bold;text-decoration: underline;">ET1705-70</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 tonsil tissue using anti-CCL5 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 (ET1705-70, 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 placenta tissue with Rabbit anti-CCL5 antibody (<a href="/products/ET1705-70" style="font-weight: bold;text-decoration: underline;">ET1705-70</a>) at 1/50 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1705-70" style="font-weight: bold;text-decoration: underline;">ET1705-70</a>) at 1/50 dilution for 1 hour 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 placenta tissue with Rabbit anti-CCL5 antibody (ET1705-70) at 1/50 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-70) at 1/50 dilution for 1 hour 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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Citation