Anti-CD147 antibody [MEM-M6/1] (ab666)
Key features and details
- Mouse monoclonal [MEM-M6/1] to CD147
- Suitable for: IHC-P, Flow Cyt, WB
- Knockout validated
- Reacts with: Human
- Isotype: IgG1
Overview
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Product name
Anti-CD147 antibody [MEM-M6/1]
See all CD147 primary antibodies -
Description
Mouse monoclonal [MEM-M6/1] to CD147 -
Host species
Mouse -
Specificity
Human CD147 antigen. This antibody recognizes an epitope in the N-terminal Ig domain (D1). This high-affinity antibody is capable of binding to unstimulated peripheral blood T cells. -
Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt HumanIHC-P HumanWB Human -
Immunogen
Recombinant full length protein corresponding to Human CD147. Purified soluble recombinant form of CD147, CD147Rg, which consists of the cDNA coding for the hinge region, CH2-and CH3 domain of human IgG1 (CD147Rg is secreted by transfectants as a dimer).
Database link: P35613 -
Positive control
- This antibody gave a positive result in IHC in the following FFPE tissue: Human normal heart muscle. Flow Cytometry: A549 cells and Peripheral blood lymphocytes. WB: A549, Raji and Jurkat.
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General notes
This product was changed from ascites to tissue culture supernatant on 24th January 2018. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.40
Preservative: 0.097% Sodium azide
Constituent: PBS -
Concentration information loading...
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Purity
Protein A purified -
Purification notes
Purified from TCS. Purity >95% by SDS-PAGE. -
Clonality
Monoclonal -
Clone number
MEM-M6/1 -
Isotype
IgG1 -
Research areas
Images
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IHC image of CD147 staining in human normal heart muscle formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 minutes. The section was then incubated with ab666, 5 µg/ml, for 15 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
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All lanes : Anti-CD147 antibody [MEM-M6/1] (ab666) at 1 µg/ml
Lane 1 : Wild-type A549 cell lysate
Lane 2 : BSG knockout A549 cell lysate
Lane 3 : Raji cell lysate
Lane 4 : Jurkat cell lysate
Lysates/proteins at 30 µg per lane.
Performed under reducing conditions.
Observed band size: 55-70 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab666 observed at 55-70 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.
ab666 was shown to react with CD147 in wild-type A549 cells in western blot with loss of signal observed in BSG knockout cell line ab273748 (knockout cell lysate ab275500). Wild-type and BSG knockout A549 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab666 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4°C at 1 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Flow cytometry overlay histogram showing wild-type A549 (green line) and BSG knockout A549 cells (ab273748) stained with ab666 (red line). The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab666) (1x106 in 100μl at 10 μg/ml) for 30 min at 4°C.
The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor® 488, pre-adsorbed) (ab150117) was used at 1/2000 for 30 min at 4°C.
Isotype control antibody was mouse IgG1κ (ab170190) used at the same concentration and conditions as the primary antibody (wild-type A549 - black line; BSG knockout A549 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
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Flow cytometry of human peripheral blood cells with ab666 at 1 µg/ml
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Overlay histogram showing peripheral blood lymphocytes stained with ab666 (red line). The cells were incubated with the antibody (ab666, 1 µg/1x106 cells) for 30 minutes at 4ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 minutes at 4ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2 µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed gating on peripheral blood lymphocytes.