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Rabbit Recombinant Monoclonal MIP-1 alpha/CCL3 antibody. Carrier free. Suitable for WB, Flow Cyt (Intra), IP, ICC/IF and reacts with Mouse, Human, Recombinant full length protein - Human, Recombinant full length protein - Mouse samples.

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Images

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Constituents: PBS

Form

Liquid

Clonality

Monoclonal

Immunogen

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

Select an application
Product promiseTestedExpectedPredictedNot recommended
WBFlow Cyt (Intra)IPICC/IFIHC-P
Human
Tested
Tested
Tested
Tested
Not recommended
Mouse
Tested
Expected
Expected
Expected
Not recommended
Recombinant full length protein - Human
Tested
Not recommended
Not recommended
Not recommended
Not recommended
Recombinant full length protein - Mouse
Tested
Not recommended
Not recommended
Not recommended
Not recommended

Tested
Tested

Species

Mouse, Human, Recombinant full length protein - Human, Recombinant full length protein - Mouse

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Expected
Expected

Species

Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Recombinant full length protein - Human, Recombinant full length protein - Mouse

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Expected
Expected

Species

Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Recombinant full length protein - Human, Recombinant full length protein - Mouse

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Expected
Expected

Species

Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Recombinant full length protein - Human, Recombinant full length protein - Mouse

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Human, Mouse, Recombinant full length protein - Human, Recombinant full length protein - Mouse

Dilution info

-

Notes

-

Target data

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).

Alternative names

Recommended products

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Rabbit Recombinant Monoclonal MIP-1 alpha/CCL3 antibody. Carrier free. Suitable for WB, Flow Cyt (Intra), IP, ICC/IF and reacts with Mouse, Human, Recombinant full length protein - Human, Recombinant full length protein - Mouse samples.

Alternative names

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

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

Yes

Clone number

EPR22529-19

Purification technique

Affinity purification Protein A

Concentration
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Storage

Shipped at conditions

Blue Ice

Appropriate long-term storage conditions

+4°C

Storage information

Do Not Freeze

Notes

ab254289 is the carrier-free version of ab229900.

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.

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.

Supplementary info

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.

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.

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

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.

5 product images

  • Western blot - Anti-MIP-1 alpha/CCL3 + CCL3L1 antibody [EPR22529-19] - BSA and Azide free (ab254289), expandable thumbnail

    Western blot - Anti-MIP-1 alpha/CCL3 + CCL3L1 antibody [EPR22529-19] - BSA and Azide free (ab254289)

    Blocking / Dilution buffer and concentration: 5% NFDM/TBST

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab229900).

    All lanes: Western blot - Anti-MIP-1 alpha/CCL3 + CCL3L1 antibody [EPR22529-19] (AB229900) at 1/1000 dilution

    Lane 1: Untreated RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 20 µg

    Lane 2: RAW 264.7 treated with 100 ng/ml LPS for 3 hours and then 300 ng/ml Brefeldin A was added for the last 3 hours, whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051) at 1/20000 dilution

    Observed band size: 12 kDa

  • Immunoprecipitation - Anti-MIP-1 alpha/CCL3 + CCL3L1 antibody [EPR22529-19] - BSA and Azide free (ab254289), expandable thumbnail

    Immunoprecipitation - Anti-MIP-1 alpha/CCL3 + CCL3L1 antibody [EPR22529-19] - BSA and Azide free (ab254289)

    Macrophage Inflammatory Protein 1 alpha / CCL3 was immunoprecipitated from 0.35 mg of THP-1 (human monocytic leukemia cell line) (treated with Phorbol-12-myristate-13-acetate (100ng/ml) for 56h, followed by adding Lipopolysaccharide (1ug/ml) for a further 16h ) whole cell lysate with ab229900 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab229900 at 1/1000 dilution.

    VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as secondary antibody at 1/5000 dilution.

    Lane 1: THP-1 (human monocytic leukemia cell line) (treated with Phorbol-12-myristate-13-acetate (100ng/ml) for 56h, followed by adding Lipopolysaccharide (1ug/ml) for a further 16h ) whole cell lysate 10 μg (Input).
    Lane 2: ab229900 IP in THP-1 (treated as above) whole lysate.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab229900 in THP-1 (treated as above) whole cell lysate.

    Blocking and dilution buffer and concentration: NFDM/TBST.
    Exposure time: 40 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab229900).

    All lanes: Immunoprecipitation - Anti-MIP-1 alpha/CCL3 + CCL3L1 antibody [EPR22529-19] (AB229900)

    Observed band size: 12 kDa

  • Western blot - Anti-MIP-1 alpha/CCL3 + CCL3L1 antibody [EPR22529-19] - BSA and Azide free (ab254289), expandable thumbnail

    Western blot - Anti-MIP-1 alpha/CCL3 + CCL3L1 antibody [EPR22529-19] - BSA and Azide free (ab254289)

    Exposure time:
    Lane 1-5 20 seconds ; Lane 6-7 3 seconds

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab229900).

    All lanes: Western blot - Anti-MIP-1 alpha/CCL3 + CCL3L1 antibody [EPR22529-19] (AB229900) at 1/1000 dilution

    Lane 1: His-tagged mouse CCL3 Recombinant Protein

    Lane 2: His-tagged human CCL3 Recombinant Protein

    Lane 3: His-tagged human CCL4 Recombinant Protein

    Lanes 4 - 5: His-tagged human CCL18 Recombinant Protein

    Lane 6: GST-tagged human CCL3L1 Recombinant Protein

    Lane 7: GST-tagged human CCL4L1 Recombinant Protein

  • Immunocytochemistry/ Immunofluorescence - Anti-MIP-1 alpha/CCL3 + CCL3L1 antibody [EPR22529-19] - BSA and Azide free (ab254289), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-MIP-1 alpha/CCL3 + CCL3L1 antibody [EPR22529-19] - BSA and Azide free (ab254289)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized THP-1 (human monocytic leukemia cell line) cells labeling Macrophage Inflammatory Protein 1 alpha / CCL3 with ab229900 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining in THP-1 treated with Phorbol-12-myristate-13-acetate (100ng/ml) for 56 h, followed by adding Lipopolysaccharide (1ug/ml) for a further 16 h. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    Treated: Cells treated with Phorbol-12-myristate-13-acetate (100ng/ml) for 56 h, followed by adding Lipopolysaccharide
    (1ug/ml) for a further 16 h.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab229900).

  • Flow Cytometry (Intracellular) - Anti-MIP-1 alpha/CCL3 + CCL3L1 antibody [EPR22529-19] - BSA and Azide free (ab254289), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-MIP-1 alpha/CCL3 + CCL3L1 antibody [EPR22529-19] - BSA and Azide free (ab254289)

    Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized THP-1 (human monocytic leukemia cell line) (untreated, green) / (treated with Phorbol-12-myristate-13-acetate (100ng/ml) for 56h, followed by adding Lipopolysaccharide (1ug/ml) for a further 16h, red) cells labeling Macrophage Inflammatory Protein 1 alpha / CCL3 with ab229900 at 1/500 dilution compared with a Rabbit monoclonal IgG Isotype control details (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG Fc (Alexa Fluor® 488) preadsorbed (ab150097), at 1/5000 dilution was used as the secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab229900).

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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