Recombinant Anti-Claudin 5 antibody [EPR7583] (ab131259)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR7583] to Claudin 5
- Suitable for: ICC/IF, WB, IHC-P
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-Claudin 5 antibody [EPR7583]
See all Claudin 5 primary antibodies -
Description
Rabbit monoclonal [EPR7583] to Claudin 5 -
Host species
Rabbit -
Specificity
ab131259 failed to detect target band in mouse brain tissue.
Mouse species is recommended based on WB result. This antibody is unsuitable to be used in IHC on mouse tissues.
This antibody failed to get signal in cell lines when performing western blot, so we do not recommend to use it when testing cell lines in westen blot.
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Tested applications
Suitable for: ICC/IF, WB, IHC-Pmore details
Unsuitable for: IHC-Fr -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Synthetic peptide within Human Claudin 5 aa 200 to the C-terminus (intracellular). The exact sequence is proprietary.
Database link: O00501 -
Positive control
- WB: Mouse lung lysates, Human fetal brain, lung squamous cell cancer and small cell lung cancer, IHC-P: Human colon, tonsil, placenta, uterus, kidney and lung tissues. ICC/IF: HUVEC 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.
Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.
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. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR7583 -
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
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab131259 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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ICC/IF | (1) |
Use a concentration of 0.008 µg/ml.
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WB | (1) |
1/1000 - 1/10000. Detects a band of approximately 23 kDa (predicted molecular weight: 23 kDa).
ab131259 failed to detect target band in mouse brain tissue. This antibody failed to get signal in cell lines when performing western blot, so we do not recommend to use it when testing cell lines in westen blot. |
IHC-P | (4) |
1/100 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Notes |
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ICC/IF
Use a concentration of 0.008 µg/ml. |
WB
1/1000 - 1/10000. Detects a band of approximately 23 kDa (predicted molecular weight: 23 kDa). ab131259 failed to detect target band in mouse brain tissue. This antibody failed to get signal in cell lines when performing western blot, so we do not recommend to use it when testing cell lines in westen blot. |
IHC-P
1/100 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Target
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Function
Plays a major role in tight junction-specific obliteration of the intercellular space. -
Sequence similarities
Belongs to the claudin family. -
Cellular localization
Cell junction > tight junction. Cell membrane. - Information by UniProt
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Database links
- Entrez Gene: 7122 Human
- Entrez Gene: 12741 Mouse
- Omim: 602101 Human
- SwissProt: O00501 Human
- SwissProt: O54942 Mouse
- Unigene: 505337 Human
- Unigene: 22768 Mouse
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Alternative names
- Androgen withdrawal and apoptosis induced protein RVP1 like antibody
- AWAL antibody
- BEC 1 antibody
see all
Images
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ab131259 staining Claudin 5 in HUV-EC cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab131259 at 0.008µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
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All lanes : Anti-Claudin 5 antibody [EPR7583] (ab131259) at 1/1000 dilution
Lane 1 : Mouse lung lysates, boiled
Lane 2 : Mouse lung lysates, unboiled
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 23 kDa
Observed band size: 20 kDa why is the actual band size different from the predicted?
Exposure time: 20 secondsWe recommend not to boil the samples after lysis to get desired WB results.
Blocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM/TBST
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All lanes : Anti-Claudin 5 antibody [EPR7583] (ab131259) at 1/1000 dilution
Lane 1 : Mouse lung tissue lysates with 5% NFDM/TBST
Lane 2 : Rat lung tissue lysates with 5% NFDM/TBST
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Predicted band size: 23 kDa
Observed band size: 20 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesWe didn’t recommend rat for that ab131259 failed to detect target band in rat lung in which Claudin 5 is highly expressed.
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All lanes : Anti-Claudin 5 antibody [EPR7583] (ab131259) at 1/1000 dilution (unpurified)
Lane 1 : Human fetal brain lysate
Lane 2 : Human lung squamous cell cancer lysate
Lane 3 : Human small cell lung cancer lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled Goat anti Rabbit IgG at 1/2000 dilution
Predicted band size: 23 kDa -
Anti-Claudin 5 antibody [EPR7583] (ab131259) at 1/5000 dilution (purified) + Human small cell lung cancer at 10 µg
Secondary
HRP goat anti-rabbit IgG (H+L) at 1/5000 dilution
Predicted band size: 23 kDa
Observed band size: 20 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST -
Anti-Claudin 5 antibody [EPR7583] (ab131259) at 1/20000 dilution (purified) + Human lung squamous cell cancer lysate at 10 µg
Secondary
HRP goat anti-rabbit IgG (H+L) at 1/5000 dilution
Predicted band size: 23 kDa
Observed band size: 20 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST -
Immunohistochemical staining of paraffin embedded human colon with purified ab131259 at a working dilution of 1/1000. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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Immunohistochemical analysis of paraffin embedded Human kidney tissue labelling Claudin 5 with unpurified ab131259 antibody at 1/100 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin embedded Human lung tissue labelling Claudin 5 with unpurified ab131259 antibody at 1/100 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Unpurified ab131259 showing positive staining in Normal tonsil tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Unpurified ab131259 showing positive staining in Normal placenta tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Unpurified ab131259 showing positive staining in Normal uterus tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Unpurified ab131259 showing positive staining in Normal colon tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (66)
ab131259 has been referenced in 66 publications.
- Yong W et al. Sodium butyrate alleviates pre-eclampsia in pregnant rats by improving the gut microbiota and short-chain fatty acid metabolites production. J Appl Microbiol 132:1370-1383 (2022). PubMed: 34470077
- Park TI et al. Routine culture and study of adult human brain cells from neurosurgical specimens. Nat Protoc 17:190-221 (2022). PubMed: 35022619
- Li Y et al. Donepezil ameliorates oxygen‑glucose deprivation/reoxygenation‑induced cardiac microvascular endothelial cell dysfunction through PARP1/NF‑κB signaling. Mol Med Rep 25:N/A (2022). PubMed: 35147204
- Cao J et al. Glaucocalyxin A alleviates lipopolysaccharide-induced inflammation and apoptosis in pulmonary microvascular endothelial cells and permeability injury by inhibiting STAT3 signaling. Exp Ther Med 23:313 (2022). PubMed: 35369532
- Li DK et al. Exosomal HMGA2 protein from EBV-positive NPC cells destroys vascular endothelial barriers and induces endothelial-to-mesenchymal transition to promote metastasis. Cancer Gene Ther 29:1439-1451 (2022). PubMed: 35388172