Anti-MCP1 antibody [RM1100] (ab315478)

Anti-MCP1 antibody [RM1100] (ab315478) is a rabbit recombinant multiclonal antibody that is used to detect MCP1 in Western Blot, IP, IHC-P, ICC/IF. Suitable for Human, Mouse samples.

- Specificity confirmed with MCP1 knockout cell line validation

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Images

Western blot - Anti-MCP1 antibody [RM1100] (AB315478), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Multiclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	Flow Cyt (Intra)	WB	IHC-P	ICC/IF	IP
Human	Not recommended	Tested	Tested	Tested	Tested
Mouse	Not recommended	Tested	Tested	Tested	Tested
Rat	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human, Mouse, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	Notes -
Species Mouse	Dilution info 1/1000	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/3000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Mouse	Dilution info 1/3000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100	Notes -
Species Mouse	Dilution info 1/100	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/30	Notes -
Species Mouse	Dilution info 1/30	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Associated Products

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Target data

See full target information CCL2

Function

Acts as a ligand for C-C chemokine receptor CCR2 (PubMed:10529171, PubMed:10587439, PubMed:9837883). Signals through binding and activation of CCR2 and induces a strong chemotactic response and mobilization of intracellular calcium ions (PubMed:10587439, PubMed:9837883). Exhibits a chemotactic activity for monocytes and basophils but not neutrophils or eosinophils (PubMed:8195247, PubMed:8627182, PubMed:9792674). May be involved in the recruitment of monocytes into the arterial wall during the disease process of atherosclerosis (PubMed:8107690).

Alternative names

MCP1, SCYA2, CCL2, C-C motif chemokine 2, HC11, Monocyte chemoattractant protein 1, Monocyte chemotactic and activating factor, Monocyte chemotactic protein 1, Monocyte secretory protein JE, Small-inducible cytokine A2, MCAF, MCP-1

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Multiclonal
Immunogens: The exact immunogen used to generate this antibody is proprietary information.
Clone number: RM1100
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

What is this antibody validated in?

Anti-MCP1 antibody [RM1100] (ab315478) is a rabbit recombinant recombinant multiclonal antibody and is validated for use in Western Blot (WB), Immunoprecipitation (IP), Immunohistochemistry (IHC-P), Immunocytochemistry/immunofluorescence (ICC/IF) in Human, Mouse samples.

What is the molecular weight of MCP1?

Anti-MCP1 [RM1100] (ab315478) specifically detects a band for MCP1 (UniProt: P13500) at a molecular weight of 11kDa.

Trial sizes available!

Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies.

Specificity confirmed

The specificity of Anti-MCP1 antibody [RM1100] (ab315478) has been confirmed by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) testing in CCL2 Knockout HeLa cells.

Other related products

We have a range of other formats of antibody clone [RM1100] also available for your convenience:
ab315478, Carrier free - ab315479

What are recombinant multiclonals?
Recombinant multiclonals are a mixture of recombinant antibodies co-expressed from a library of heavy and light chains. They offer several advantages including:

- The sensitivity of polyclonal antibodies by recognising multiple epitopes
- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

View our range of recombinant multiclonal antibodies.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

MCP-1 also known as CCL2 is a chemokine involved in the recruitment of monocytes to sites of injury and inflammation. This protein consists of 76 amino acids and has a molecular weight of about 11 kDa. MCP-1 is expressed in a wide range of cell types including endothelial cells fibroblasts and monocytes. It acts by binding to the chemokine receptor CCR2 stimulating intracellular signaling pathways that mediate cell migration.

Biological function summary

The role of MCP-1 is significant in guiding immune cells to areas requiring immune response. It does not form part of a complex but operates as a singular entity to affect immune signaling. MCP-1 influences the behavior of monocytes and macrophages by inducing chemotaxis and activating their effector functions. Through this mechanism it aids in the inflammatory response and tissue repair.

Pathways

MCP-1 participates in the inflammatory pathways by cooperating with key proteins like CCR2. It facilitates the JAK-STAT signaling pathway which is vital for transmitting extracellular information into cellular responses. The interaction of MCP-1 with CCR2 influences the activation of downstream proteins like STAT3 enabling various immune responses. MCP-1 thereby plays a central role in modulating inflammation-related pathways.

Associated diseases and disorders

MCP-1 has a connection to inflammatory bowel disease and atherosclerosis. In such conditions elevated levels of MCP-1 can exacerbate inflammatory responses by recruiting monocytes to dysfunctional sites worsening tissue damage. Through its interaction with CCR2 MCP-1 becomes a pivotal factor in chronic inflammation. Targeting this MCP-1/CCR2 axis can provide therapeutic opportunities for modulating disease progression in these inflammatory diseases.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

13 product images

Western blot - Anti-MCP1 antibody [RM1100] (ab315478)
MCP1 Western blot staining using rabbit Anti-MCP1 antibody
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-MCP1 antibody [RM1100] (ab315478) at 1/1000 dilution
Lane 1: Untreated THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg
Lane 2: THP-1 treated with 80nM Phorbol-12-myristate-13-acetate (PMA) for 24 hours, then added 100ng/ml Lipopolysaccharides (LPS) for 7 hours, 1µg/ml Brefeldin A was added for additional 3 hours whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 11 kDa, 36 kDa
Exposure time: 26s
Western blot - Anti-MCP1 antibody [RM1100] (ab315478)
MCP1 Western blot staining using rabbit Anti-MCP1 antibody
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 28955950).
Panel A is applied with Anti-MCP1 antibody [RM1100] (ab315478) at 1/1000 dilution and panel B is applied with Anti-MCP1 antibody (Anti-MCP1 antibody ab25124) at 1/1000 dilution.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
Exposure time: 26 seconds (panel A);180 seconds (panel B).
All lanes: Western blot - Anti-MCP1 antibody [RM1100] (ab315478) at 1/1000 dilution
Lane 1: Untreated RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg
Lane 2: RAW 264.7 treated with 100ng/ml Lipopolysaccharides (LPS) for 7 hours, then 1µg/ml Brefeldin A was added for additional 3 hours whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 20 kDa, 36 kDa
Immunoprecipitation - Anti-MCP1 antibody [RM1100] (ab315478)
MCP1 was immunoprecipitated from 0.35 mg RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) treated with 100ng/ml Lipopolysaccharides (LPS) for 7 hours, then 1µg/ml Brefeldin A was added for additional 3 hours whole cell lysate with ab315478 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab315478 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) treated with 100ng/ml Lipopolysaccharides (LPS) for 7 hours, then 1µg/ml Brefeldin A was added for additional 3 hours whole cell lysate

Lane 2: ab315478 IP in RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) treated with 100ng/ml Lipopolysaccharides (LPS) for 7 hours, then 1µg/ml Brefeldin A was added for additional 3 hours whole cell lysate

Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab315478 in RAW 264.7 treated with 100ng/ml Lipopolysaccharides (LPS) for 7 hours, then 1µg/ml Brefeldin A was added for additional 3 hours whole cell lysate
All lanes: Immunoprecipitation - Anti-MCP1 antibody [RM1100] (ab315478) at 1/30 dilution
All lanes: RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) treated with 100ng/ml Lipopolysaccharides (LPS) for 7 hours, then 1µg/ml Brefeldin A was added for additional 3 hours whole cell lysate
Secondary
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Exposure time: 15s
Immunoprecipitation - Anti-MCP1 antibody [RM1100] (ab315478)
MCP1 was immunoprecipitated from 0.35 mg THP-1 (human monocytic leukemia monocyte) treated with 80nM Phorbol-12-myristate-13-acetate (PMA) for 24 hours, then added 100ng/ml Lipopolysaccharides (LPS) for 7 hours, 1µg/ml Brefeldin A was added for last 3 hours whole cell lysate with ab315478 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab315478 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: THP-1 (human monocytic leukemia monocyte) treated with 80nM Phorbol-12-myristate-13-acetate (PMA) for 24 hours, then added 100ng/ml Lipopolysaccharides (LPS) for 7 hours, 1µg/ml Brefeldin A was added for last 3 hours whole cell lysate

Lane 2: ab315478 IP in THP-1 (human monocytic leukemia monocyte) treated with 80nM Phorbol-12-myristate-13-acetate (PMA) for 24 hours, then added 100ng/ml Lipopolysaccharides (LPS) for 7 hours, 1µg/ml Brefeldin A was added for last 3 hours whole cell lysate

Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab315478 in THP-1 treated with 80nM Phorbol-12-myristate-13-acetate (PMA) for 24 hours, then added 100ng/ml Lipopolysaccharides (LPS) for 7 hours, 1µg/ml Brefeldin A was added for last 3 hours whole cell lysate
All lanes: Immunoprecipitation - Anti-MCP1 antibody [RM1100] (ab315478) at 1/30 dilution
All lanes: THP-1 (human monocytic leukemia monocyte) treated with 80nM Phorbol-12-myristate-13-acetate (PMA) for 24 hours, then added 100ng/ml Lipopolysaccharides (LPS) for 7 hours, 1µg/ml Brefeldin A was added for last 3 hours whole cell lysate
Secondary
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Exposure time: 180s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MCP1 antibody [RM1100] (ab315478)
Immunohistochemical analysis of paraffin-embedded: Image A - RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) cell pellets treated with 100ng/ml Lipopolysaccharides (LPS) for 7 hours, then 1µg/ml Brefeldin A was added for additional 3 hours; Image B - Untreated Raw264.7 cell pellets tissue labeling MCP1 with ab315478 at 1/3000 (0.172 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Cytoplasmic staining on RAW 264.7 cell pellets treated with 100ng/ml Lipopolysaccharides (LPS) for 7 hours, then 1µg/ml Brefeldin A was added for additional 3 hours (Image A); No staining on untreated RAW 264.7 cell pellets (Images B).

The images on the left are applied with Anti-MCP1 antibody [RM1100] (ab315478) at 1/3000 dilution and the images on the right are applied with Anti-MCP1 antibody (Anti-MCP1 antibody ab25124) at 1/1500 dilution.

The section was incubated with ab315478 or Anti-MCP1 antibody ab25124 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Western blot - Anti-MCP1 antibody [RM1100] (ab315478)
Western blot: Anti-MCP1 antibody (ab315478) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5](Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red.
In Western blot, ab315478 was shown to bind specifically to MCP1. Target of interest was observed at 11 kDa in wild-type HeLa cell lysates (lane 1/2) with no signal observed at this size in MCP1 knockout cell line (lane 3/4, knockout cell line Human CCL2 (MCP1) knockout HeLa cell line ab255372/knockout cell lysate ab263807). To generate this image, samples were first run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in a fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed in TBS-T, incubated with secondary antibodies Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution for 1 h at room temperature, washed again then imaged.
All lanes: Western blot - Anti-MCP1 antibody [RM1100] (ab315478) at 1/1000 dilution
Lane 1: Wild-type HeLa (human cervical adenocarcinoma epithelial cell) treated with 1µg/ml Brefeldin A for 3 hours whole cell lysate at 20 µg with ½ volume of Odyssey Blocking Buffer (TBS)+ ½ volume of TBS
Lane 2: Wild-type HeLa treated with 1µg/ml TNF-alpha (Recombinant human TNF alpha protein ab9642) for 6 hours, 1µg/ml Brefeldin A was added for additional 3 hours whole cell lysate at 20 µg with ½ volume of Odyssey Blocking Buffer (TBS)+ ½ volume of TBS
Lane 3: CCL2 knockout HeLa treated with 1µg/ml Brefeldin A for 3 hours whole cell lysate at 20 µg with ½ volume of Odyssey Blocking Buffer (TBS)+ ½ volume of TBS
Lane 4: CCL2 knockout HeLa treated with TNF-alpha (Recombinant human TNF alpha protein ab9642) for 6 hours, 1µg/ml Brefeldin A was added for additional 3 hours whole cell lysate at 20 µg with ½ volume of Odyssey Blocking Buffer (TBS)+ ½ volume of TBS
Secondary
All lanes: Goat Anti-Rabbit IgG H&L (800CW) and Goat Anti-Mouse IgG H&L (680RD) at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 11 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MCP1 antibody [RM1100] (ab315478)
Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling MCP1 with ab315478 at 1/3000 (0.172 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Negative control: ab315478 showed no staining on mouse skeletal muscle (ab), while Anti-MCP1 antibody ab25124 showed non-specific staining on the mouse skeletal muscle.

The image on the left is applied with Anti-MCP1 antibody [RM1100] (ab315478) at 1/3000 dilution and the image on the right is applied with Anti-MCP1 antibody (Anti-MCP1 antibody ab25124) at 1/1500 dilution.

The section was incubated with ab315478 or Anti-MCP1 antibody ab25124 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MCP1 antibody [RM1100] (ab315478)
Immunohistochemical analysis of paraffin-embedded: Image A - Mouse lung treated with 1ug/ml Lipopolysaccharides (LPS) and 1ug/ml Brefeldin A for 16 hours; Image B - Untreated mouse lung tissue labeling MCP1 with ab315478 at 1/3000 (0.172 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Cytoplasmic staining on mouse lung treated with 1ug/ml Lipopolysaccharides (LPS) and 1ug/ml Brefeldin A for 16 hours (Image A); nearly no satining on untreated mouse lung (Image B).

The images on the left are applied with Anti-MCP1 antibody [RM1100] (ab315478) at 1/3000 dilution and the images on the right are applied with Anti-MCP1 antibody (Anti-MCP1 antibody ab25124) at 1/1500 dilution.

The section was incubated with ab315478 or Anti-MCP1 antibody ab25124 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MCP1 antibody [RM1100] (ab315478)
Immunohistochemical analysis of paraffin-embedded: Image A - Wild-type Hela cell pellets treated with Brefeldin A (1ug/ml) for 3 hours; Image B - Wild-type Hela cell pellets treated with TNF-alpha (TNFα, 20ng/ml) for 3 hours, 1µg/ml Brefeldin A was added for additional 3 hours; Image C - CCL2 knockout HeLa cell pellets treated with Brefeldin A (1ug/ml) for 3 hours; Image D - CCL2 knockout HeLa cell pellets treated with TNF-alpha (TNFα, 20ng/ml) for 3 hours, 1µg/ml Brefeldin A was added for additional 3 hours tissue labeling MCP1 with ab315478 at 1/3000 (0.172 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on Wild-type Hela cell pellets treated with Brefeldin A (1ug/ml) for 3 hours (Image A) and Wild-type Hela cell pellets treated with TNF-alpha (TNFα, 20ng/ml) for 3 hours, 1µg/ml Brefeldin A was added for additional 3 hours (Image B); No staining on CCL2 knockout HeLa cell pellets treated with Brefeldin A (1ug/ml) for 3 hours (Image C) and CCL2 knockout HeLa cell pellets treated with TNF-alpha (TNFα, 20ng/ml) for 3 hours, 1µg/ml Brefeldin A was added for additional 3 hours (Image D).

The section was incubated with ab315478 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MCP1 antibody [RM1100] (ab315478)
Immunohistochemical analysis of paraffin-embedded: Image A - THP-1 (human monocytic leukemia monocyte) cell pellets treated with 80nM Phorbol-12-myristate-13-acetate (PMA) for 24 hours, then added 100ng/ml Lipopolysaccharides (LPS) for 7 hours, 1µg/ml Brefeldin A was added for additional 3 hours; Image B - Untreated THP-1 cell pellets tissue labeling MCP1 with ab315478 at 1/3000 (0.172 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Cytoplasmic staining on THP-1 cell pellets treated with 80nM Phorbol-12-myristate-13-acetate (PMA) for 24 hours, then added 100ng/ml Lipopolysaccharides (LPS) for 7 hours, 1µg/ml Brefeldin A was added for additional 3 hours (Image A); No staining on untreated THP-1 cell pellets (Image B).

The section was incubated with ab315478 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunocytochemistry/ Immunofluorescence - Anti-MCP1 antibody [RM1100] (ab315478)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) cells labelling MCP1 with ab315478 at 1/100 (5.17 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).
Confocal image showing cytoplasmic staining on RAW 264.7 cells treated with 100ng/ml Lipopolysaccharides (LPS) for 7 hours, then 1µg/ml Brefeldin A was added for additional 3 hours; no staining on untreated RAW 264.7 cells.

Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).
Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.
Immunocytochemistry/ Immunofluorescence - Anti-MCP1 antibody [RM1100] (ab315478)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized THP-1 (human monocytic leukemia monocyte) cells labelling MCP1 with ab315478 at 1/100 (5.17 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).
Confocal image showing cytoplasmic staining on THP-1 cells treated with 80nM Phorbol-12-myristate-13-acetate (PMA) for 24 hours, then added 100ng/ml Lipopolysaccharides (LPS) for 7 hours, 1µg/ml Brefeldin A was added for additional 3 hours; no staining on untreated THP-1 cells.

Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).
Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MCP1 antibody [RM1100] (ab315478)
Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling MCP1 with ab315478 at 1/3000 (0.172 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Negative control: no staining on human skeletal muscle.

The section was incubated with ab315478 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins