Anti-MCP3 antibody [EPR22649-155] - BSA and Azide free (ab256821)
![Anti-MCP3 antibody [EPR22649-155] - BSA and Azide free (ab256821)](https://www.abcam.com/ps/products/256/ab256821/Images/ab256821-450555-anti-mcp3-antibody-epr22649-155-bsa-and-azide-free-western-blot-wildtype-raw-264-7-untreated-control-wildtype-raw-264-7-pma-treated-80.jpg "Anti-MCP3 antibody [EPR22649-155] - BSA and Azide free (ab256821)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22649-155] to MCP3 - BSA and Azide free
- Suitable for: WB, IP
- Knockout validated
- Reacts with: Mouse
Overview
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Product name
Anti-MCP3 antibody [EPR22649-155] - BSA and Azide free
See all MCP3 primary antibodies -
Description
Rabbit monoclonal [EPR22649-155] to MCP3 - BSA and Azide free -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species IP MouseWB Mouse
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Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: RAW264.7 (treated with 80 nM PMA), whole cell lysate; HEK-293T transfected with MCP3 expression vector containing a myc-His-tag®, whole lysate. IP: RAW264.7 (treated with 80 nM PMA for 24 hours, then with 100 ng/ml LPS treated for 6 hours with addition of 1ug/ml of Brefeldin A for the last 3 hours), cell lysate.
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General notes
Ab256821 is the carrier-free version of ab228979. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.
Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with
ab256821 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.
Maxpar® is a trademark of Fluidigm Canada Inc.
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. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22649-155 -
Isotype
IgG -
Research areas
Images
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Western blot - Anti-MCP3 antibody [EPR22649-155] - BSA and Azide free (ab256821)All lanes : Anti-MCP3 antibody [EPR22649-155] (ab228979) at 1/1000 dilution
Lane 1 : Wild-type RAW 264.7 untreated control cell lysate
Lane 2 : Wild-type RAW 264.7 PMA treated (80 nM, 24 h) plus LPS treated (100 ng/ml, 6 h) and Brefeldin A (ab120299) treated (5 µg/ml, 5 h) cell lysate
Lane 3 : CCL7/MCP3 knockout RAW 264.7 untreated cell lysate
Lane 4 : CCL7/MCP3 knockout RAW 264.7 PMA treated (80 nM, 24 h) plus LPS treated (100 ng/ml, 6 h) and Brefeldin A (ab120299) treated (5 µg/ml, 5 h) cell lysate
Lane 5 : J774A.1 Glucose treated (138.8 mMol/L, 8 h) plus Brefeldin A treated (5 µg/ml, 6 h) and Brefeldin A (ab120299) treated (5 µg/ml, 5 h) cell lysate
Lane 6 : J774A.1 Glucose treated (5.6 mMol/L, 8 h) plus Brefeldin A treated (5 µg/ml, 6 h) and Brefeldin A (ab120299) treated (5 µg/ml, 5 h) cell lysate
Lysates/proteins at 30 µg per lane.
Performed under reducing conditions.
Predicted band size: 11 kDa
Observed band size: 14 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation (ab228979).
Lanes 1 - 6: Merged signal (red and green). Green - ab228979 observed at 14 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab228979 was shown to react with MCP3 in RAW 264.7 wild-type cells in Western blot with loss of signal observed in CCL7 knockout sample. Wild-type and CCL7 knockout RAW 264.7 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab228979 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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![Immunoprecipitation - Anti-MCP3 antibody [EPR22649-155] - BSA and Azide free (ab256821)](https://www.abcam.com/ps/products/256/ab256821/Images/ab256821-2-anti-mcp3-antibody-epr22649-155-immunoprecipitation-raw-264-7-mouse.jpg)
Immunoprecipitation - Anti-MCP3 antibody [EPR22649-155] - BSA and Azide free (ab256821)MCP3 was immunoprecipitated from 0.35 mg RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) (treated with 80 nM Phorbol-12-myristate-13-acetate (PMA) for 24 hours, then with 100 ng/ml lipopolysaccharides (LPS) treated for 6 hours with addition of 1ug/ml of Brefeldin A for the last 3 hours), whole cell lysate with ab228979 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab228979 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.
Lane 1: RAW264.7 (treated as above) whole cell lysate 10ug
Lane 2: ab228979 IP in RAW264.7 (treated as above) whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab228979 in RAW264.7 (treated as above) whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 min.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab228979).
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![Anti-MCP3 antibody [EPR22649-155] - BSA and Azide free (ab256821)](https://www.abcam.com/ps/products/256/ab256821/Images/ab256821-3-benefits-of-recombinant-antibodies.png)
Anti-MCP3 antibody [EPR22649-155] - BSA and Azide free (ab256821)
