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Rabbit Recombinant Monoclonal CCL4/MIP-1 beta antibody. Suitable for IP, ELISA, WB, ICC/IF and reacts with Human, Mouse samples. Cited in 12 publications.


Images

Western blot - Anti-CCL4/MIP-1 beta antibody [EP521Y] (AB45690), expandable thumbnail
  • Western blot - Anti-CCL4/MIP-1 beta antibody [EP521Y] (AB45690), expandable thumbnail
  • Immunoprecipitation - Anti-CCL4/MIP-1 beta antibody [EP521Y] (AB45690), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-CCL4/MIP-1 beta antibody [EP521Y] (AB45690), expandable thumbnail
  • Western blot - Anti-CCL4/MIP-1 beta antibody [EP521Y] (AB45690), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 50% Glycerol (glycerin, glycerine), 49% PBS, 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

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

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPELISAWBICC/IFFlow Cyt (Intra)
Human
Tested
Tested
Tested
Tested
Not recommended
Mouse
Expected
Expected
Tested
Expected
Not recommended

Tested
Tested

Species
Human
Dilution info
1/60
Notes

-

Expected
Expected

Species
Mouse
Dilution info
Use at an assay dependent concentration.
Notes

-

Tested
Tested

Species
Human
Dilution info
-
Notes

-

Expected
Expected

Species
Mouse
Dilution info
Use at an assay dependent concentration.
Notes

-

Tested
Tested

Species
Mouse
Dilution info
1/1000 - 1/500000
Notes

-

Species
Human
Dilution info
1/1000 - 1/500000
Notes

-

Tested
Tested

Species
Human
Dilution info
1/100
Notes

-

Expected
Expected

Species
Mouse
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Human, Mouse
Dilution info
-
Notes

-

Associated Products

Select an associated product type

5 products for Alternative Product

Target data

Function

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.

Alternative names

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

Recommended products

Rabbit Recombinant Monoclonal CCL4/MIP-1 beta antibody. Suitable for IP, ELISA, WB, ICC/IF and reacts with Human, Mouse samples. Cited in 12 publications.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EP521Y
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
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

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:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

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

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.

Biological function summary

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.

Pathways

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.

Associated diseases and disorders

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.

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.

7 product images

  • Western blot - Anti-CCL4/MIP-1 beta antibody [EP521Y] (ab45690), expandable thumbnail

    Western blot - Anti-CCL4/MIP-1 beta antibody [EP521Y] (ab45690)

    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, 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] (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

  • Western blot - Anti-CCL4/MIP-1 beta antibody [EP521Y] (ab45690), expandable thumbnail

    Western blot - Anti-CCL4/MIP-1 beta antibody [EP521Y] (ab45690)

    Blocking/Diluting buffer and concentration 5% NFDM /TBST

    All lanes: Western blot - 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

    Secondary

    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

  • Immunoprecipitation - Anti-CCL4/MIP-1 beta antibody [EP521Y] (ab45690), expandable thumbnail

    Immunoprecipitation - Anti-CCL4/MIP-1 beta antibody [EP521Y] (ab45690)

    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 (+): 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 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, 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/Diluting buffer 5% NFDM/TBST

    All lanes: Immunoprecipitation - Anti-CCL4/MIP-1 beta antibody [EP521Y] (ab45690)

    Predicted band size: 10 kDa

    Exposure time: 3min

  • Immunocytochemistry/ Immunofluorescence - Anti-CCL4/MIP-1 beta antibody [EP521Y] (ab45690), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-CCL4/MIP-1 beta antibody [EP521Y] (ab45690)

    Immunocytochemistry/Immunofluorescence analysis of THP-1 (Human monocytic leukemia cell line) cells labeling CCL4/MIP-1 beta + CCL4L with 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.

  • Western blot - Anti-CCL4/MIP-1 beta antibody [EP521Y] (ab45690), expandable thumbnail

    Western blot - Anti-CCL4/MIP-1 beta antibody [EP521Y] (ab45690)

    Blocking/Diluting buffer and concentration 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] (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

    Secondary

    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

  • Western blot - Anti-CCL4/MIP-1 beta antibody [EP521Y] (ab45690), expandable thumbnail

    Western blot - Anti-CCL4/MIP-1 beta antibody [EP521Y] (ab45690)

    Blocking/Diluting buffer and concentration 5% NFDM /TBST

    All lanes: Western blot - 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

    Secondary

    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

  • ELISA - Anti-CCL4/MIP-1 beta antibody [EP521Y] (ab45690), expandable thumbnail

    ELISA - Anti-CCL4/MIP-1 beta antibody [EP521Y] (ab45690)

    ELISA analysis of Human CCL4/MIP-1 beta recombinant protein at 500 ng/mL with 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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Product protocols

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