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CCL2 / MCP-1 Recombinant Rabbit Monoclonal Antibody [PSH18-16] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA751662

CCL2 / MCP-1 Recombinant Rabbit Monoclonal Antibody [PSH18-16] - BSA and Azide free

Application

  • WB

  • IF-Cell

  • FC

Reactivity

  • Mouse

  • Rat

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

CCL2 / MCP-1 Recombinant Rabbit Monoclonal Antibody [PSH18-16] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within rat CCL2 aa 1-148.

Species Reactivity

Mouse, Rat

Validated Applications

WB, IF-Cell, FC

Molecular Weight

Predicted band size: 16 kDa

Positive Control

RAW264.7 treated with 100nM TPA overnight then treated with 100ng/mL LPS for 7 hours add 1μg/mL BFA for last 3 hours cell lysate, NR8383 treated with 100nM TPA overnight then treated with 100ng/mL LPS for 7 hours add 1μg/mL BFA for last 3 hours cell lysate.

Conjugation

unconjugated

Clone Number

PSH18-16

Product Features

Form

Liquid

Concentration

1mg/ml

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

  • 1:100-1:250

  • 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[5] 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. Ryu S et al. Peripheral CCL2-CCR2 signalling contributes to chronic headache-related sensitization. Brain. 2023 Oct

2. Georgakis MK et al. Targeting the CCL2-CCR2 axis for atheroprotection. Eur Heart J. 2022 May

Subcellular Location

Secreted.

UNIPROT

SwissProt: P10148 Mouse

SwissProt: P14844 Rat

Synonyms

C-C motif chemokine 2

Immediate-early serum-responsive protein JE

Monocyte chemoattractant protein 1

Monocyte chemotactic protein 1

MCP-1

Small-inducible cytokine A2

Ccl2

Je

Mcp1

Scya2

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of CCL2 / MCP-1 on different lysates with Rabbit anti-CCL2 / MCP-1 antibody (<a href="/products/HA751662" style="font-weight: bold;text-decoration: underline;">HA751662</a>) at 1/5,000 dilution.<br /><br />Lane 1: RAW264.7 cell lysate (20 µg/Lane)<br />Lane 2: RAW264.7 treated with 100nM TPA overnight then treated with 100ng/mL LPS for 7 hours add 1μg/mL BFA for last 3 hours cell lysate (20 µg/Lane)<br /><br />Predicted band size: 16 kDa<br />Observed band size: 20 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/HA751662" style="font-weight: bold;text-decoration: underline;">HA751662</a>) at 1/5,000 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.

    ☑ Cell treatment (CT)

    Western blot analysis of CCL2 / MCP-1 on different lysates with Rabbit anti-CCL2 / MCP-1 antibody (HA751662) at 1/5,000 dilution.

    Lane 1: RAW264.7 cell lysate (20 µg/Lane)
    Lane 2: RAW264.7 treated with 100nM TPA overnight then treated with 100ng/mL LPS for 7 hours add 1μg/mL BFA for last 3 hours cell lysate (20 µg/Lane)

    Predicted band size: 16 kDa
    Observed band size: 20 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 (HA751662) at 1/5,000 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.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of CCL2 / MCP-1 on different lysates with Rabbit anti-CCL2 / MCP-1 antibody (<a href="/products/HA751662" style="font-weight: bold;text-decoration: underline;">HA751662</a>) at 1/2,000 dilution.<br /><br />Lane 1: NR8383 cell lysate (40 µg/Lane)<br />Lane 2: NR8383 treated with 100nM TPA overnight then treated with 100ng/mL LPS for 7 hours add 1μg/mL BFA for last 3 hours cell lysate (40 µg/Lane)<br /><br />Predicted band size: 16 kDa<br />Observed band size: 20 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/HA751662" style="font-weight: bold;text-decoration: underline;">HA751662</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.

    ☑ Cell treatment (CT)

    Western blot analysis of CCL2 / MCP-1 on different lysates with Rabbit anti-CCL2 / MCP-1 antibody (HA751662) at 1/2,000 dilution.

    Lane 1: NR8383 cell lysate (40 µg/Lane)
    Lane 2: NR8383 treated with 100nM TPA overnight then treated with 100ng/mL LPS for 7 hours add 1μg/mL BFA for last 3 hours cell lysate (40 µg/Lane)

    Predicted band size: 16 kDa
    Observed band size: 20 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 (HA751662) 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.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Immunocytochemistry analysis of RAW264.7 cells untreated / treated with 100nM TPA overnight then treated with 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/HA751662" style="font-weight: bold;text-decoration: underline;">HA751662</a>) at 1/250 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/HA751662" style="font-weight: bold;text-decoration: underline;">HA751662</a>) at 1/250 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.

    ☑ Cell treatment (CT)

    Immunocytochemistry analysis of RAW264.7 cells untreated / treated with 100nM TPA overnight then treated with 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 (HA751662) at 1/250 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 (HA751662) at 1/250 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.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Immunocytochemistry analysis of NR8383 cells untreated / treated with 100nM TPA overnight then treated with 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/HA751662" style="font-weight: bold;text-decoration: underline;">HA751662</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/HA751662" style="font-weight: bold;text-decoration: underline;">HA751662</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.

    ☑ Cell treatment (CT)

    Immunocytochemistry analysis of NR8383 cells untreated / treated with 100nM TPA overnight then treated with 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 (HA751662) 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 (HA751662) 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.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Flow cytometric analysis of RAW264.7 cells untreated (left) / treated with 100nM TPA overnight then treated with 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/HA751662" style="font-weight: bold;text-decoration: underline;">HA751662</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).

    ☑ Cell treatment (CT)

    Flow cytometric analysis of RAW264.7 cells untreated (left) / treated with 100nM TPA overnight then treated with 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 (HA751662, 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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