Anti-CCL3 / MIP-1 alpha + CCL3L1 antibody [EPR23751-54] - BSA and Azide
- BOND RX™ Validated
- RabMAb
- Recombinant
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(1 Publication)
Rabbit Recombinant Monoclonal MIP-1 alpha/CCL3 antibody. Carrier free. Suitable for ICC/IF, IHC-P, IP, WB, Flow Cyt (Intra) and reacts with Mouse, Human, Recombinant fragment - Human samples. Cited in 1 publication.
View 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
- Flow Cyt (Intra)
Supplier Data
Flow Cytometry (Intracellular) - Anti-CCL3 / MIP-1 alpha + CCL3L1 antibody [EPR23751-54] - BSA and Azide (AB277944)
This data was developed using ab259372, the same antibody clone in a different buffer formulation.
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 (ab172730) (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor®488, ab150077), at 1/2000 dilution was used as the secondary antibody.
- ICC/IF
Supplier Data
Immunocytochemistry/ Immunofluorescence - Anti-CCL3 / MIP-1 alpha + CCL3L1 antibody [EPR23751-54] - BSA and Azide (AB277944)
This data was developed using ab259372, the same antibody clone in a different buffer formulation.
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 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 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 ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) secondary antibody at 1/1000 dilution.
- WB
Supplier Data
Western blot - Anti-CCL3 / MIP-1 alpha + CCL3L1 antibody [EPR23751-54] - BSA and Azide (AB277944)
This data was developed using ab259372, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer : 5% NFDM/TBST.
Exposure time : 8 seconds.
All lanes:
Western blot - Anti-CCL3 / MIP-1 alpha + CCL3L1 antibody [EPR23751-54] (<a href='/en-us/products/primary-antibodies/ccl3-mip-1-alpha-ccl3l1-antibody-epr23751-54-ab259372'>ab259372</a>) 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 (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution
Observed band size: 12 kDa
false
- Flow Cyt (Intra)
Supplier Data
Flow Cytometry (Intracellular) - Anti-CCL3 / MIP-1 alpha + CCL3L1 antibody [EPR23751-54] - BSA and Azide (AB277944)
This data was developed using ab259372, the same antibody clone in a different buffer formulation.
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 (ab172730) (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor®488, ab150077), at 1/2000 dilution was used as the secondary antibody.
- IHC-P
Supplier Data
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CCL3 / MIP-1 alpha + CCL3L1 antibody [EPR23751-54] - BSA and Azide (AB277944)
This data was developed using ab259372, the same antibody clone in a different buffer formulation.
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 (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 (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
- IHC-P
Supplier Data
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CCL3 / MIP-1 alpha + CCL3L1 antibody [EPR23751-54] - BSA and Azide (AB277944)
This data was developed using ab259372, the same antibody clone in a different buffer formulation.
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 (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 (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
- IHC-P
Supplier Data
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CCL3 / MIP-1 alpha + CCL3L1 antibody [EPR23751-54] - BSA and Azide (AB277944)
This data was developed using ab259372, the same antibody clone in a different buffer formulation.
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 (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 (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
- ICC/IF
Supplier Data
Immunocytochemistry/ Immunofluorescence - Anti-CCL3 / MIP-1 alpha + CCL3L1 antibody [EPR23751-54] - BSA and Azide (AB277944)
This data was developed using ab259372, the same antibody clone in a different buffer formulation.
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 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 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 ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) secondary antibody at 1/1000 dilution.
- IHC-P
Supplier Data
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CCL3 / MIP-1 alpha + CCL3L1 antibody [EPR23751-54] - BSA and Azide (AB277944)
This data was developed using ab259372, the same antibody clone in a different buffer formulation.
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 (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 (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
- IP
Supplier Data
Immunoprecipitation - Anti-CCL3 / MIP-1 alpha + CCL3L1 antibody [EPR23751-54] - BSA and Azide (AB277944)
This data was developed using ab259372, the same antibody clone in a different buffer formulation.
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)(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 (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-CCL3 / MIP-1 alpha + CCL3L1 antibody [EPR23751-54] (<a href='/en-us/products/primary-antibodies/ccl3-mip-1-alpha-ccl3l1-antibody-epr23751-54-ab259372'>ab259372</a>)
Observed band size: 10 kDa
false
- WB
Supplier Data
Western blot - Anti-CCL3 / MIP-1 alpha + CCL3L1 antibody [EPR23751-54] - BSA and Azide (AB277944)
This data was developed using ab259372, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer : 5% NFDM/TBST.
Exposire time : 10 seconds.
All lanes:
Western blot - Anti-CCL3 / MIP-1 alpha + CCL3L1 antibody [EPR23751-54] (<a href='/en-us/products/primary-antibodies/ccl3-mip-1-alpha-ccl3l1-antibody-epr23751-54-ab259372'>ab259372</a>) 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 (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution
Observed band size: 12 kDa
false
- WB
Supplier Data
Western blot - Anti-CCL3 / MIP-1 alpha + CCL3L1 antibody [EPR23751-54] - BSA and Azide (AB277944)
This data was developed using ab259372, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer : 5% NFDM/TBST.
Exposure time : 3 seconds.
All lanes:
Western blot - Anti-CCL3 / MIP-1 alpha + CCL3L1 antibody [EPR23751-54] (<a href='/en-us/products/primary-antibodies/ccl3-mip-1-alpha-ccl3l1-antibody-epr23751-54-ab259372'>ab259372</a>) 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) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution
Observed band size: 12 kDa
false
Related conjugates and formulations (1)
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Anti-CCL3 / MIP-1 alpha + CCL3L1 antibody [EPR23751-54]
Reactivity data
Product details
ab277944 is the carrier-free version of ab259372.
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.
What are the advantages of a recombinant monoclonal antibody?
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.
Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. 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.
Compatibility
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.
Properties and storage information
Form
Purification technique
Storage buffer
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Product protocols
- Visit the General protocols
- Visit the Troubleshooting
Target data
Additional targets
Publications (1)
Recent publications for all applications. Explore the full list and refine your search
BMC medicine 22:11 PubMed38185631
2024
Applications
Unspecified application
Species
Unspecified reactive species
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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