Recombinant Anti-MIP-1 alpha/CCL3 + CCL3L1 antibody [EPR22529-19] (ab229900)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22529-19] to MIP-1 alpha/CCL3 + CCL3L1
- Suitable for: Flow Cyt (Intra), WB, ICC/IF, IP
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-MIP-1 alpha/CCL3 + CCL3L1 antibody [EPR22529-19]
See all MIP-1 alpha/CCL3+CCL3L1 primary antibodies -
Description
Rabbit monoclonal [EPR22529-19] to MIP-1 alpha/CCL3 + CCL3L1 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, ICC/IF, IPmore details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: THP-1 (treated with PMA and LPS) whole cell lysate, RAW 264.7(treated with LPS and BFA) whole cell lysate IP: THP-1 (treated with PMA and LPS) whole cell lysate. ICC/IF: THP-1 (treated with PMA and LPS) cells. Flow Cyt (intra): THP-1 (treated with PMA and LPS) cells.
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General notes
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22529-19 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab229900 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Flow Cyt (Intra) |
1/500.
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WB |
1/1000. Predicted molecular weight: 10 kDa.
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ICC/IF |
1/100.
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IP |
1/30.
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Notes |
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Flow Cyt (Intra)
1/500. |
WB
1/1000. Predicted molecular weight: 10 kDa. |
ICC/IF
1/100. |
IP
1/30. |
Target
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Cellular localization
MIP-1 alpha/CCL3: Secreted. CCL3L1: Secreted. -
Database links
- Entrez Gene: 414062 Human
- Entrez Gene: 6348 Human
- Entrez Gene: 6349 Human
- Entrez Gene: 20302 Mouse
- Omim: 182283 Human
- Omim: 609468 Human
- SwissProt: P10147 Human
- SwissProt: P16619 Human
see all
Images
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All lanes : 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
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
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 10 kDa
Observed band size: 12 kDa why is the actual band size different from the predicted?Blocking / Dilution buffer and concentration: 5% NFDM/TBST
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All lanes : 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
Predicted band size: 10 kDaExposure time:
Lane 1-5 20 seconds ; Lane 6-7 3 seconds -
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. -
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. -
All lanes : Anti-MIP-1 alpha/CCL3 + CCL3L1 antibody [EPR22529-19] (ab229900) at 1/1000 dilution
Lane 1 : Untreated THP-1 (human monocytic leukemia cell line) whole cell lysate
Lane 2 : THP-1 (treated with 100 ng/ml phorbol-12-myristate-13-acetate (PMA) for 56 hours, followed by adding 1 µg/ml lipopolysaccharide (LPS) for a further 16 hours ) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 10 kDa
Observed band size: 12 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking and dilution buffer: 5% NFDM/TBST.
This blot was developed using a higher sensitivity ECL substrate.
The molecular weight observed is consistent with what has been described in the literature (PMID: 11369791)
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Intracellular flow cytometric analysis of4% 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.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (1)
ab229900 has been referenced in 1 publication.
- Yu W et al. miR-4456/CCL3/CCR5 Pathway in the Pathogenesis of Tight Junction Impairment in Chronic Obstructive Pulmonary Disease. Front Pharmacol 12:551839 (2021). PubMed: 33953665