Rabbit Recombinant Monoclonal CCL4/MIP-1 beta antibody. Carrier free. Suitable for ICC/IF, IP, ELISA, WB and reacts with Human, Mouse samples.
pH: 7.2 - 7.4
Constituents: PBS
ICC/IF | IP | ELISA | WB | |
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Human | Tested | Tested | Tested | Tested |
Mouse | Expected | Expected | Expected | Tested |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes - |
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Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
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Species Mouse, Human | Dilution info - | Notes - |
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Monokine with inflammatory and chemokinetic properties. Binds to CCR5. One of the major HIV-suppressive factors produced by CD8+ T-cells. Recombinant MIP-1-beta induces a dose-dependent inhibition of different strains of HIV-1, HIV-2, and simian immunodeficiency virus (SIV). The processed form MIP-1-beta(3-69) retains the abilities to induce down-modulation of surface expression of the chemokine receptor CCR5 and to inhibit the CCR5-mediated entry of HIV-1 in T-cells. MIP-1-beta(3-69) is also a ligand for CCR1 and CCR2 isoform B.
LAG1, MIP1B, SCYA4, CCL4, C-C motif chemokine 4, G-26 T-lymphocyte-secreted protein, HC21, Lymphocyte activation gene 1 protein, MIP-1-beta(1-69), Macrophage inflammatory protein 1-beta, PAT 744, Protein H400, SIS-gamma, Small-inducible cytokine A4, T-cell activation protein 2, LAG-1, MIP-1-beta, ACT-2
Rabbit Recombinant Monoclonal CCL4/MIP-1 beta antibody. Carrier free. Suitable for ICC/IF, IP, ELISA, WB and reacts with Human, Mouse samples.
pH: 7.2 - 7.4
Constituents: PBS
ab187674 is the carrier-free version of Anti-CCL4/MIP-1 beta antibody [EP521Y] ab45690.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
CCL4 also known as MIP-1 beta or MIP-1b has a mechanical role as a chemokine involved in immune response regulation. The protein has a mass of about 7.8 kDa. It expresses in various tissues including leukocytes where it mainly functions as a chemoattractant. MIP-1 beta helps recruit immune cells like natural killer cells and monocytes to sites of inflammation or injury. Researchers frequently measure its levels using assays like MIP-1 beta ELISA or CCL4 ELISA.
CCL4 is critical in immune system modulation. It coordinates leukocyte activation and trafficking contributing to inflammation and immune surveillance. It does not function in isolation but rather interacts with other chemokines and receptors. CCL4 alongside its sibling chemokines forms a dynamic network that ensures a balanced immune response. It connects to receptors such as CCR5 playing substantial roles in its activities.
CCL4 participates in essential signaling cascades that mediate immune responses. It is notably part of the chemokine signaling pathway which is instrumental in directing cell movement. CCL4 also engages with proteins like CCR5 in the inflammatory and immune response pathways. This interaction enhances communication between cells required for effective pathogen defense.
CCL4 plays roles in immune-related conditions. It is strongly linked to inflammatory diseases such as rheumatoid arthritis and HIV infection. In rheumatoid arthritis CCL4 levels are often elevated correlating with inflammation severity. During HIV infection CCL4 interacts with proteins like CCR5 a co-receptor the virus uses to enter host cells. This involvement makes MIP-1 beta a target for studies seeking therapeutic interventions in these diseases.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our 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.
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False colour image of Western blot: Anti-CCL4/MIP-1 beta antibody [EP521Y] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-CCL4/MIP-1 beta antibody [EP521Y] ab45690 was shown to bind specifically to CCL4/MIP-1 beta. A band was observed at 12 kDa in wild-type THP-1 cell lysates with no signal observed at this size in CCL4 knockout cell line Human CCL4 knockout THP-1 cell line ab273719 (knockout cell lysate Human CCL4 knockout THP-1 cell lysate ab275512). To generate this image, wild-type and CCL4 knockout THP-1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-CCL4/MIP-1 beta antibody [EP521Y] (Anti-CCL4/MIP-1 beta antibody [EP521Y] ab45690) at 1/1000 dilution
Lane 1: Wild-type THP-1 Vehicle control + Brefeldin A (5 u/mL, 6 h) cell lysate at 20 µg
Lane 2: Wild-type THP-1 Treated PMA (100 ng/mL, 56 h) + LPS (1 u/mL, 24 h) + Brefeldin A (5 u/mL, 6 h) cell lysate at 20 µg
Lane 2: Western blot - Human CCL4 knockout THP-1 cell line (Human CCL4 knockout THP-1 cell line ab273719)
Lane 3: CCL4 knockout THP-1 Vehicle control + Brefeldin A (5 u/mL, 6 h) cell lysate at 20 µg
Lane 4: CCL4 knockout THP-1 Treated PMA (100 ng/mL, 56 h) + LPS (1 u/mL, 24 h) + Brefeldin A (5 u/mL, 6 h) cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 10 kDa
Observed band size: 12 kDa
This data was developed using Anti-CCL4/MIP-1 beta antibody [EP521Y] ab45690, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-CCL4/MIP-1 beta antibody [EP521Y] (Anti-CCL4/MIP-1 beta antibody [EP521Y] ab45690) at 1/1000 dilution
Lane 1: Untagged human CCL4 recombinant protein (aa24-92) at 10 µg
Lane 2: Untagged human CCL4L recombinant protein (aa24-92) at 10 µg
Lane 3: GST-tagged human CCL3 recombinant protein (aa27-92) at 10 µg
Lane 4: GST-tagged human CCL3L recombinant protein 2*(aa28-93) at 10 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 10 kDa
Observed band size: 12 kDa
Exposure time: 5s
This data was developed using Anti-CCL4/MIP-1 beta antibody [EP521Y] ab45690, the same antibody clone in a different buffer formulation.
Anti-CCL4/MIP-1 beta antibody [EP521Y] ab45690 at 1/60 immunoprecipitating CCL4/MIP-1 beta + CCL4L in THP-1 (Human monocytic leukemia cell line) whole cell lysate observed at 12 KDa (lanes 1 and 2).
Lane 1 (input): THP-1 treated with 100 nM PMA overnight, then treated with 100 ng/mL LPS for 7 hours and 1 μg/mL Brefeldin A was added for the last 3 hours whole cell lysate, 10μg.
Lane 2 (+): Anti-CCL4/MIP-1 beta antibody [EP521Y] ab45690 + THP-1 treated with 100 nM PMA overnight, then treated with 100 ng/mL LPS for 7 hours and 1 μg/mL Brefeldin A was added for the last 3 hours whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-CCL4/MIP-1 beta antibody [EP521Y] ab45690 in THP-1 treated with 100 nM PMA overnight, then treated with 100 ng/mL LPS for 7 hours and 1 μg/mL Brefeldin A was added for the last 3 hours whole cell lysate.
For western blotting, Anti-CCL4/MIP-1 beta antibody [EP521Y] ab45690 at 1/1000 and VeriBlot for IP Detection Reagent (HRP) ab131366 VeriBlot for IP Detection Reagent (HRP) was used for detection (1/1000).
Blocking and dilution buffer: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-CCL4/MIP-1 beta antibody [EP521Y] (Anti-CCL4/MIP-1 beta antibody [EP521Y] ab45690)
Predicted band size: 10 kDa
Exposure time: 3min
This data was developed using Anti-CCL4/MIP-1 beta antibody [EP521Y] ab45690, the same antibody clone in a different buffer formulation.Immunocytochemistry/Immunofluorescence analysis of THP-1 (Human monocytic leukemia cell line) cells labeling CCL4/MIP-1 beta + CCL4L with Anti-CCL4/MIP-1 beta antibody [EP521Y] ab45690 at 1/100. Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were counter-stained with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution. DAPI was used to stain nuclei blue. The expression increased after treatment with Lipopolysaccharides (LPS), 100 ng/mL for 4 hours, followed by addition of Brefeldin A (1 μg/mL) for 3 hours.
This data was developed using Anti-CCL4/MIP-1 beta antibody [EP521Y] ab45690, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
CCL4/MIP-1 beta is induced in macrophages following exposure to bacterial LPS (PMID: 9848081).
All lanes: Western blot - Anti-CCL4/MIP-1 beta antibody [EP521Y] (Anti-CCL4/MIP-1 beta antibody [EP521Y] ab45690) at 1/1000 dilution
Lane 1: Untreated THP-1 (human acute monocytic leukemia) cell lysate at 10 µg
Lane 2: THP-1 treated with 100 nM Phorbol-12-myristate-13-acetate(PMA) overnight, then treated with Lipopolysaccharides (LPS) 100 ng/mL for 7 hours and then 1 µg/mL Brefeldin A was added for the last 3 hours, lysate at 10 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051)
Predicted band size: 10 kDa
Observed band size: 12 kDa
Exposure time: 3min
This data was developed using Anti-CCL4/MIP-1 beta antibody [EP521Y] ab45690, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-CCL4/MIP-1 beta antibody [EP521Y] (Anti-CCL4/MIP-1 beta antibody [EP521Y] ab45690) at 1/1000 dilution
Lane 1: Untreated Raw264.7 (mouse abelson murine leukemia virus-induced tumor) whole cell lysate at 10 µg
Lane 2: Raw264.7 (mouse abelson murine leukemia virus-induced tumor) treated with LPS 10μg/mL for 4 hours and then 1 μg/mL Brefeldin A was added for the last 3 hours lysate at 10 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 10 kDa, 18 kDa, 25 kDa, 55 kDa, 57 kDa
Observed band size: 12 kDa, 18 kDa, 26 kDa, 90 kDa
Exposure time: 3min
This data was developed using Anti-CCL4/MIP-1 beta antibody [EP521Y] ab45690, the same antibody clone in a different buffer formulation.ELISA analysis of Human CCL4/MIP-1 beta recombinant protein at 500 ng/mL with Anti-CCL4/MIP-1 beta antibody [EP521Y] ab45690. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution was used as the secondary antibody.
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