Anti-Cytosolic Phospholipase A2 antibody (ab73406)
Key features and details
- Rabbit polyclonal to Cytosolic Phospholipase A2
- Suitable for: IHC-P, WB, ICC/IF
- Knockout validated
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-Cytosolic Phospholipase A2 antibody
See all Cytosolic Phospholipase A2 primary antibodies -
Description
Rabbit polyclonal to Cytosolic Phospholipase A2 -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, WB, ICC/IFmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat, Rabbit, Horse, Cow, Dog, Orangutan
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Immunogen
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Positive control
- This antibody gave a positive signal in HeLa Whole Cell Lysate - Bleomycin Treated (40U/ml)
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
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 or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Assay kits
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab73406 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 |
|---|---|---|
| IHC-P |
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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| WB |
Use a concentration of 1 µg/ml. Detects a band of approximately 100 kDa (predicted molecular weight: 85 kDa).
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| ICC/IF |
Use a concentration of 1 µg/ml.
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| Notes |
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IHC-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
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WB
Use a concentration of 1 µg/ml. Detects a band of approximately 100 kDa (predicted molecular weight: 85 kDa). |
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ICC/IF
Use a concentration of 1 µg/ml. |
Target
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Function
Selectively hydrolyzes arachidonyl phospholipids in the sn-2 position releasing arachidonic acid. Together with its lysophospholipid activity, it is implicated in the initiation of the inflammatory response. -
Tissue specificity
Expressed in various tissues such as macrophages, platelets, neutrophils, fibroblasts and lung endothelium. -
Sequence similarities
Contains 1 C2 domain.
Contains 1 PLA2c domain. -
Domain
The N-terminal C2 domain associates with lipid membranes upon calcium binding. It modulates enzyme activity by presenting the active site to its substrate in response to elevations of cytosolic Ca(2+). -
Post-translational
modificationsActivated by phosphorylation at both Ser-505 and Ser-727. -
Cellular localization
Cytoplasm. Cytoplasmic vesicle. Translocates to membrane vesicles in a calcium-dependent fashion. - Information by UniProt
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Database links
- Entrez Gene: 525072 Cow
- Entrez Gene: 100033889 Horse
- Entrez Gene: 151056 Human
- Entrez Gene: 5321 Human
- Entrez Gene: 18783 Mouse
- Entrez Gene: 665270 Mouse
- Entrez Gene: 100173125 Orangutan
- Entrez Gene: 100008748 Rabbit
see all -
Alternative names
- Calcium dependent phospholipid binding protein antibody
- CPLA 2 antibody
- cPLA2 alpha antibody
see all
Images
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Western blot - Anti-Cytosolic Phospholipase A2 antibody (ab73406)
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: Cytosolic Phospholipase A2 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: A549 whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab73406 observed at 90 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab73406 was shown to specifically recognize Cytosolic Phospholipase A2 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when Cytosolic Phospholipase A2 knockout samples were examined. Wild-type and Cytosolic Phospholipase A2 knockout samples were subjected to SDS-PAGE. Ab73406 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/20000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour at room temperature before imaging.
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Immunocytochemistry/ Immunofluorescence - Anti-Cytosolic Phospholipase A2 antibody (ab73406)ICC/IF image of ab73406 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab73406, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) Hek293, HepG2 and MCF7 cells at 1µg/ml, and in 100% methanol fixed (5 min) HeLa cells at 1µg/ml. -
Western blot - Anti-Cytosolic Phospholipase A2 antibody (ab73406)Anti-Cytosolic Phospholipase A2 antibody (ab73406) at 1 µg/ml + HeLa Whole Cell Lysate - Bleomycin Treated (40U/ml) at 10 µg
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 85 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?The 100kDa band is comparable to molecular weights seen with other commercially available antibodies to Cytosolic Phospholipase A2
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytosolic Phospholipase A2 antibody (ab73406)IHC image of Cytosolic Phospholipsae A2 staining in human colon carcinoma 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 (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab73406, 5µg/ml, for 15 mins 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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (2)
ab73406 has been referenced in 2 publications.
- Emre C et al. Age-related changes in brain phospholipids and bioactive lipids in the APP knock-in mouse model of Alzheimer's disease. Acta Neuropathol Commun 9:116 (2021). PubMed: 34187579
- Yang Y et al. Sevoflurane attenuates ventilator-induced lung injury by regulating c-PLA2 expression. Mol Med Rep 18:2923-2928 (2018). PubMed: 30015951