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CCL2 / MCP-1 Recombinant Rabbit Monoclonal Antibody [PSH12-44] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA751533

CCL2 / MCP-1 Recombinant Rabbit Monoclonal Antibody [PSH12-44] - BSA and Azide free

Application

  • WB

  • IF-Cell

  • FC

Reactivity

  • Mouse

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

CCL2 / MCP-1 Recombinant Rabbit Monoclonal Antibody [PSH12-44] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Mouse CCL2 aa 24-148.

Species Reactivity

Mouse

Validated Applications

WB, IF-Cell, FC

Molecular Weight

Predicted band size: 16 kDa

Positive Control

RAW264.7 treated with 100ng/mL LPS for 4 hours add 1μg/mL BFA for3 hours cell lysate.

Conjugation

unconjugated

Clone Number

PSH12-44

Product Features

Form

Liquid

Concentration

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:5,000

  • IF-Cell

  • 1:100

  • FC

  • 1:1,000

Target

Function

The chemokine (C-C motif) ligand 2 (CCL2) is also referred to as monocyte chemoattractant protein 1 (MCP1) and small inducible cytokine A2. CCL2 is a small cytokine that belongs to the CC chemokine family. CCL2 tightly regulates cellular mechanics and thereby recruits monocytes, memory T cells, and dendritic cells to the sites of inflammation produced by either tissue injury or infection. CCL2 is a monomeric polypeptide, with a molecular weight of approximately 13-15 kDa depending on levels of glycosylation. CCL2 is anchored in the plasma membrane of endothelial cells by glycosaminoglycan side chains of proteoglycans. CCL2 is primarily secreted by monocytes, macrophages and dendritic cells. Platelet derived growth factor is a major inducer of CCL2 gene.

Background References

1. Xu M et al. Role of the CCL2-CCR2 signalling axis in cancer: Mechanisms and therapeutic targeting. Cell Prolif. 2021 Oct

2. Zhang H et al. Role of the CCL2-CCR2 axis in cardiovascular disease: Pathogenesis and clinical implications. Front Immunol. 2022 Aug

Subcellular Location

Secreted.

UNIPROT

SwissProt: P10148 Mouse

Synonyms

C-C motif chemokine 2 antibody

CCL2 antibody

CCL2_HUMAN antibody

Chemokine (C C motif) ligand 2 antibody

GDCF 2 antibody

GDCF-2 antibody

GDCF2 antibody

HC11 antibody

HSMCR30 antibody

JE antibody

Expand

Images

  • Western blot analysis of CCL2 / MCP-1 on different lysates with Rabbit anti-CCL2 / MCP-1 antibody (<a href="/products/HA751533" style="font-weight: bold;text-decoration: underline;">HA751533</a>) at 1/2,000 dilution.<br /><br />Lane 1: RAW264.7 cell lysate<br />Lane 2: RAW264.7 treated with 100ng/mL LPS for 4 hours add 1μg/mL BFA for3 hours cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 16 kDa<br />Observed band size: 16 kDa<br /><br />Exposure time: 3 minutes; 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/HA751533" style="font-weight: bold;text-decoration: underline;">HA751533</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.

    Western blot analysis of CCL2 / MCP-1 on different lysates with Rabbit anti-CCL2 / MCP-1 antibody (HA751533) at 1/2,000 dilution.

    Lane 1: RAW264.7 cell lysate
    Lane 2: RAW264.7 treated with 100ng/mL LPS for 4 hours add 1μg/mL BFA for3 hours cell lysate

    Lysates/proteins at 10 µg/Lane.

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

    Exposure time: 3 minutes; 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 (HA751533) 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 100ng/mL LPS for 4 hours add 1μg/mL BFA for3 hours labeling CCL2 / MCP-1 with Rabbit anti-CCL2 / MCP-1 antibody (<a href="/products/HA751533" style="font-weight: bold;text-decoration: underline;">HA751533</a>) at 1/100 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/HA751533" style="font-weight: bold;text-decoration: underline;">HA751533</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/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 100ng/mL LPS for 4 hours add 1μg/mL BFA for3 hours labeling CCL2 / MCP-1 with Rabbit anti-CCL2 / MCP-1 antibody (HA751533) at 1/100 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 (HA751533) 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 (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 100ng/mL LPS for 4 hours add 1μg/mL BFA for3 hours (right) labeling CCL2 / MCP-1.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA751533" style="font-weight: bold;text-decoration: underline;">HA751533</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 100ng/mL LPS for 4 hours add 1μg/mL BFA for3 hours (right) labeling CCL2 / MCP-1.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA751533, 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).

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