Anti-Macrophage Inflammatory Protein 1 alpha / CCL3 + CCL3L1 antibody [EPR23751-54] (ab259372)

Rabbit Recombinant Monoclonal MIP-1 alpha/CCL3 antibody. Suitable for ICC/IF, IP, WB, Flow Cyt (Intra), IHC-P and reacts with Mouse, Human, Recombinant fragment - Human samples. Cited in 3 publications.

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Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Macrophage Inflammatory Protein 1 alpha / CCL3 + CCL3L1 antibody [EPR23751-54] (AB259372), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 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

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

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/50	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/50	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Recombinant fragment - Human, Rat	Dilution info -	Notes -

Tested
Tested

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

Not recommended
Not recommended

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

Tested
Tested

Species	Dilution info	Notes
Species Recombinant fragment - Human	Dilution info 1/1000	Notes -
Species Mouse	Dilution info 1/1000	Notes -
Species Human	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 Mouse	Dilution info 1/500	Notes -
Species Human	Dilution info 1/500	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Recombinant fragment - Human, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	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 Mouse	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Recombinant fragment - Human	Dilution info -	Notes -

Target data

See full target information CCL3

Function

Monokine with inflammatory and chemokinetic properties. Binds to CCR1, CCR4 and CCR5. One of the major HIV-suppressive factors produced by CD8+ T-cells. Recombinant MIP-1-alpha induces a dose-dependent inhibition of different strains of HIV-1, HIV-2, and simian immunodeficiency virus (SIV).

Additional Targets

CCL3L1

Alternative names

G0S19-1, MIP1A, SCYA3, CCL3, C-C motif chemokine 3, G0/G1 switch regulatory protein 19-1, Macrophage inflammatory protein 1-alpha, PAT 464.1, SIS-beta, Small-inducible cytokine A3, Tonsillar lymphocyte LD78 alpha protein, MIP-1-alpha

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogens: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR23751-54
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

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

MIP-1 alpha also known as CCL3 or CCL3L1 is a chemokine with important roles in immune regulation. It has a molecular mass of approximately 7.8 kDa. This protein is expressed mainly in immune cells like macrophages T cells and natural killer (NK) cells. The expression of MIP-1 alpha occurs in response to inflammatory signals and it acts to recruit other immune cells to the site of infection or injury. Its function is essential for orchestrating the cellular movements during immune surveillance and defensive responses.

Biological function summary

The MIP-1 alpha functions as a chemotactic cytokine. It binds to receptors like CCR1 and CCR5 on the surface of target cells resulting in activation and migration toward areas of inflammation. MIP-1 alpha does not form a complex with other proteins; however its interactions with its receptors are vital for signal transduction. It also promotes the production of additional inflammatory cytokines contributing to amplifying immune responses. The chemotactic properties of MIP-1 alpha make it an important player in the body's defense against pathogens.

Pathways

MIP-1 alpha is involved in important immune pathways like the NF-kB signaling pathway. This pathway plays a significant role in immune cell activation and the inflammatory response. MIP-1 alpha interacts with other chemokines like CCL4 (MIP-1 beta) which shares similar functions and binds to CCR5. The coordinated activities of these chemokines enhance the recruitment and activation of various leukocyte populations facilitating an effective immune response.

Associated diseases and disorders

MIP-1 alpha exhibits a connection to inflammatory diseases such as rheumatoid arthritis and HIV infection. In rheumatoid arthritis MIP-1 alpha contributes to joint inflammation and damage by attracting immune cells to synovial joints. In the context of HIV it interacts with the CCR5 receptor affecting viral entry into CD4+ T cells. The inhibition of MIP-1 alpha and its interaction with CCR5 provides a therapeutic strategy in controlling HIV progression. These associations highlight the chemokine's impact on both autoimmune disorders and infectious diseases.

Product promise

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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.

12 product images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Macrophage Inflammatory Protein 1 alpha / CCL3 + CCL3L1 antibody [EPR23751-54] (ab259372)
Immunohistochemical analysis of paraffin-embedded human Hodgkin lymphoma tissue labeling Macrophage Inflammatory Protein 1 alpha / CCL3 + CCL3L1 with ab259372 at 1/1000 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Positive staining on human hodgkin Lymphoma (PMID: 31581676). The section was incubated with ab259372 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunoprecipitation - Anti-Macrophage Inflammatory Protein 1 alpha / CCL3 + CCL3L1 antibody [EPR23751-54] (ab259372)
Macrophage Inflammatory Protein 1 alpha / CCL3 + CCL3L1 was immunoprecipitated from 0.35 mg THP-1 (human monocytic leukemia monocyte) (treated with 100nM PMA for 16 hours. And then replace PMA with 100 ng/ml LPS (Lipopolysaccharide) for 7 hours, 1 μg/ml BFA was added for the last 3 hours) whole cell lysate whole cell lysate with ab259372 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab259372 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: THP-1 (treated as above) whole cell lysate 3 ug
Lane 2: ab259372 IP in THP-1 (treated as above) whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab259372 in THP-1 (treated as above) whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 8 seconds.
All lanes: Immunoprecipitation - Anti-Macrophage Inflammatory Protein 1 alpha / CCL3 + CCL3L1 antibody [EPR23751-54] (ab259372)
Observed band size: 10 kDa
Immunocytochemistry/ Immunofluorescence - Anti-Macrophage Inflammatory Protein 1 alpha / CCL3 + CCL3L1 antibody [EPR23751-54] (ab259372)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X100 permeabilized THP-1 (human monocytic leukemia monocyte) cells labeling Macrophage Inflammatory Protein 1 alpha / CCL3 + CCL3L1 with ab259372 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in THP-1 cells treated with 12-myristate 13-acetate (100 nM) for 16 hours then with lipopolysaccharide (100 ng/ml) for 4 hours and with brefeldin A (1μg/ml) for another 3 hours. Tubulin is counterstained using Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) at 1/200 dilution (Red). The nuclear counterstain is DAPI (Blue).
Secondary antibody only control: Used PBS instead of priary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) secondary antibody at 1/1000 dilution.
Flow Cytometry (Intracellular) - Anti-Macrophage Inflammatory Protein 1 alpha / CCL3 + CCL3L1 antibody [EPR23751-54] (ab259372)
Intracellular flow cytometric analysis of 5% paraformaldehyde-fixed, 90% methanol-permeabilized THP-1 (Human monocytic leukemia monocyte) (treated with 100nM phorbol 12-myristate 13-acetate (PMA) for 16 hours, then 100ng/ml lipopolysaccharide (LPS) for 4 hours, and add 1ug/ml Brefeldin A (BFA) for another 3 hours) (Red)/ Untreated control (Green) cells labeling Macrophage Inflammatory Protein 1 alpha / CCL3 + CCL3L1 with ab259372 at 1/500 dilution (Red / Green) compared with a Isotype control details (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077), at 1/2000 dilution was used as the secondary antibody.
Western blot - Anti-Macrophage Inflammatory Protein 1 alpha / CCL3 + CCL3L1 antibody [EPR23751-54] (ab259372)
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 8 seconds.
All lanes: Western blot - Anti-Macrophage Inflammatory Protein 1 alpha / CCL3 + CCL3L1 antibody [EPR23751-54] (ab259372) 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 100nM PMA for 16 hours. And then replace PMA with 100 ng/ml LPS (Lipopolysaccharide) for 4 hours, and then 1 μg/ml Brefeldin A was added for the last 3 hours), whole cell lysate at 20 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution
Observed band size: 12 kDa
Western blot - Anti-Macrophage Inflammatory Protein 1 alpha / CCL3 + CCL3L1 antibody [EPR23751-54] (ab259372)
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 3 seconds.
All lanes: Western blot - Anti-Macrophage Inflammatory Protein 1 alpha / CCL3 + CCL3L1 antibody [EPR23751-54] (ab259372) at 1/5000 dilution
Lane 1: Untreated RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg
Lane 2: RAW 264.7 (treated with 100 ng/ml LPS(Lipopolysaccharide) for 4 hours and then 1 μg/ml Brefeldin A was added for the last 3 hours), whole cell 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 1/20000 dilution
Observed band size: 12 kDa
Immunocytochemistry/ Immunofluorescence - Anti-Macrophage Inflammatory Protein 1 alpha / CCL3 + CCL3L1 antibody [EPR23751-54] (ab259372)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X100 permeabilized RAW264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) cells labeling Macrophage Inflammatory Protein 1 alpha / CCL3 + CCL3L1 with ab259372 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in RAW 264.7 cells treated with 12-myristate 13-acetate (100 nM) for 16 hours then with lipopolysaccharide (100 ng/ml) for 4 hours and with brefeldin A (1μg/ml) for another 3 hours. Tubulin is counterstained using Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) at 1/200 dilution (Red). The nuclear counterstain is DAPI (Blue).
Secondary antibody only control: Used PBS instead of priary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) secondary antibody at 1/1000 dilution.
Flow Cytometry (Intracellular) - Anti-Macrophage Inflammatory Protein 1 alpha / CCL3 + CCL3L1 antibody [EPR23751-54] (ab259372)
Intracellular flow cytometric analysis of5% paraformaldehyde-fixed, 90% methanol-permeabilized RAW264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) (treated with 100ng/ml lipopolysaccharide (LPS) for 4 hours, and add Brefeldin A (BFA) for another 3 hours (Red)/ Untreated control (Green)) (Red)/ Untreated control (Green) cells labeling Macrophage Inflammatory Protein 1 alpha / CCL3 + CCL3L1 with ab259372at 1/500 dilution (Red / Green) compared with a Isotype control details (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor^®488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077), at 1/2000 dilution was used as the secondary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Macrophage Inflammatory Protein 1 alpha / CCL3 + CCL3L1 antibody [EPR23751-54] (ab259372)
Immunohistochemical analysis of paraffin-embedded THP-1 cells labeling Macrophage Inflammatory Protein 1 alpha / CCL3 + CCL3L1 with ab259372 at 1/1000 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Positive staining on THP-1 treated with 100nM PMA, LPS and Brefeldin treatment (image A), and negative staining on untreat THP-1 (image B). The section was incubated with ab259372 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Western blot - Anti-Macrophage Inflammatory Protein 1 alpha / CCL3 + CCL3L1 antibody [EPR23751-54] (ab259372)
Blocking and dilution buffer: 5% NFDM/TBST.
Exposire time: 10 seconds.
All lanes: Western blot - Anti-Macrophage Inflammatory Protein 1 alpha / CCL3 + CCL3L1 antibody [EPR23751-54] (ab259372) at 1/1000 dilution
Lane 1: His-tagged mouse CCL3 recombinant protein (aa24-92) * 2 ,10 ng
Lane 2: GST/His-tagged human CCL3L1 recombinant protein (aa24-93), 10 ng
Lane 3: LIF/His-tagged human CCL4 recombinant protein (aa24-92)*2, 10 ng
Lane 4: His-tagged human CCL18 recombinant protein (aa21-89),10 ng
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution
Observed band size: 12 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Macrophage Inflammatory Protein 1 alpha / CCL3 + CCL3L1 antibody [EPR23751-54] (ab259372)
Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling Macrophage Inflammatory Protein 1 alpha / CCL3 + CCL3L1 with ab259372 at 1/1000 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Negative control: No staining on human tonsil. The section was incubated with ab259372 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Macrophage Inflammatory Protein 1 alpha / CCL3 + CCL3L1 antibody [EPR23751-54] (ab259372)
Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling Macrophage Inflammatory Protein 1 alpha / CCL3 + CCL3L1 with ab259372 at 1/1000 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Negative control: No staining on human spleen tissue. The section was incubated with ab259372 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.