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CCL2 / MCP-1 Rabbit Polyclonal Antibody

Usd: 315

Specification

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Catalog# HA500267

CCL2 / MCP-1 Rabbit Polyclonal Antibody

Application

  • WB

  • IF-Cell

  • FC

Reactivity

  • Mouse

Conjugation

  • unconjugated

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

Overview

Product Name

CCL2 / MCP-1 Rabbit Polyclonal Antibody

Antibody Type

Rabbit Polyclonal Antibody

Immunogen

Recombinant protein within rat CCL2 aa 1-148.

Species Reactivity

Mouse

Validated Applications

WB, IF-Cell, FC

Molecular Weight

Predicted band size: 16 kDa

Positive Control

RAW264.7 treated with 100nM TPA overnight then 100ng/mL LPS for 7 hours add 1μg/mL BFA for last 3 hours cell lysate, RAW264.7 cells treated with 100nM TPA overnight then 100ng/mL LPS for 7 hours add 1μg/mL BFA for last 3 hours.

Conjugation

unconjugated

RRID

AB_3071363

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

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Immunogen affinity purified.

Application Dilution

  • WB

  • 1:5,000

  • IF-Cell

  • 1:500

  • FC

  • 1:1,000

Target

Function

CCL2 exhibits a chemotactic activity for monocytes and basophils. However, it does not attract neutrophils or eosinophils. After deletion of the N-terminal residue, CCL2 loses its attractivity for basophils and becomes a chemoattractant of eosinophils. Basophils and mast cells that are treated with CCL2 release their granules to the intercellular space. This effect can be also potentiated by a pre-treatment with IL-3 or even by other cytokines. CCL2 augments monocyte anti-tumor activity and it is essential for formation of granulomas. CCL2 protein become a CCR2 antagonist when it is cleaved by metalloproteinase MMP-12.

Background References

1. Li Y.S. et. al. The expression of monocyte chemotactic protein (MCP-1) in human vascular endothelium in vitro and in vivo. Mol. Cell. Biochem. 126:61-68(1993).

Subcellular Location

Secreted.

UNIPROT

SwissProt: P10148 Mouse

Synonyms

C-C motif chemokine 2 antibody

CCL2 antibody

Chemokine (C C motif) ligand 2 antibody

GDCF 2 antibody

GDCF-2 antibody

GDCF2 antibody

HC11 antibody

HSMCR30 antibody

JE antibody

MCAF antibody

Expand

Images

  • Western blot analysis of CCL2 / MCP-1 on different lysates with Rabbit anti-CCL2 / MCP-1 antibody (<a href="/products/HA500267" style="font-weight: bold;text-decoration: underline;">HA500267</a>) at 1/5,000 dilution.<br /><br />Lane 1: RAW264.7 cell lysate<br />Lane 2: RAW264.7 treated with 100nM TPA overnight then 100ng/mL LPS for 7 hours add 1μg/mL BFA for last 3 hours cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 16 kDa<br />Observed band size: 16 kDa<br /><br />Exposure time: 10 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/HA500267" style="font-weight: bold;text-decoration: underline;">HA500267</a>) at 1/5,000 dilution was used in primary antibody dilution (K1803) 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 CCL2 / MCP-1 on different lysates with Rabbit anti-CCL2 / MCP-1 antibody (HA500267) at 1/5,000 dilution.

    Lane 1: RAW264.7 cell lysate
    Lane 2: RAW264.7 treated with 100nM TPA overnight then 100ng/mL LPS for 7 hours add 1μg/mL BFA for last 3 hours cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 16 kDa
    Observed band size: 16 kDa

    Exposure time: 10 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 (HA500267) 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.

  • Immunocytochemistry analysis of RAW264.7 cells untreated / treated with 100nM TPA overnight then 100ng/mL LPS for 7 hours add 1μg/mL BFA for last 3 hours labeling CCL2 / MCP-1 with Rabbit anti-CCL2 / MCP-1 antibody (<a href="/products/HA500267" style="font-weight: bold;text-decoration: underline;">HA500267</a>) at 1/500 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-CCL2 / MCP-1 antibody (<a href="/products/HA500267" style="font-weight: bold;text-decoration: underline;">HA500267</a>) at 1/500 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/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</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 RAW264.7 cells untreated / treated with 100nM TPA overnight then 100ng/mL LPS for 7 hours add 1μg/mL BFA for last 3 hours labeling CCL2 / MCP-1 with Rabbit anti-CCL2 / MCP-1 antibody (HA500267) at 1/500 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-CCL2 / MCP-1 antibody (HA500267) at 1/500 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 (HA601187, 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.

  • Flow cytometric analysis of RAW264.7 cells untreated (left) / treated with 100nM TPA overnight then 100ng/mL LPS for 7 hours add 1μg/mL BFA for last 3 hours (right) labeling CCL2 / MCP-1.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA500267" style="font-weight: bold;text-decoration: underline;">HA500267</a>, 1/1,000) (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 RAW264.7 cells untreated (left) / treated with 100nM TPA overnight then 100ng/mL LPS for 7 hours add 1μg/mL BFA for last 3 hours (right) labeling CCL2 / MCP-1.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA500267, 1/1,000) (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).

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

Alternative Products

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CCL2 / MCP-1 Recombinant Rabbit Monoclonal Antibody [PSH12-44]

Application: WB,IF-Cell,FC

Reactivity: Mouse

Conjugate: unconjugated