Anti-Occludin 抗体abcam *（货号ab31721）新年*5折*-上海博耀生物科技有限公司手机版

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Product nameAnti-Occludin抗体
参阅全部 Occludin 一抗
Description
兔多克隆抗体to Occludin
SpecificityThe immunogen used to raise this antibody has 78% homology with the Mouse Occludin protein. Some customers have successfully used ab31721 with Mouse lysates/tissue. Please contact Abcam Scientific Support for more information.
Tested applicationsIHC-FoFr, WB, In-Cell ELISA, ICC/IF, IHC-Pmore details
Species reactivity
Reacts with: Rat, Human, Pig
Predicted to work with: Dog
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 350 - 450 of Human Occludin.
参阅Abcam的专有抗源政策

(Peptide available as ab34440.)
Positive control
- ab31721 gave a positive signal in Colon (Human), Liver (Human), and Kidney (Human) Tissue Lysates.

Properties

FormLiquid
Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration100 ?g at 1 mg/ml
PurityImmunogen affinity purified
ClonalityPolyclonal
IsotypeIgG
Research Areas
- Signal Transduction
- Cytoskeleton / ECM
- Cell Adhesion
- Tight Junctions

Applications

Our Abpromise guarantee covers the use of ab31721 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-FoFr		Use at an assay dependent concentration. PubMed: 20978075
WB		Use a concentration of 1 ?g/ml. Detects a band of approximay 63 kDa (predicted molecular weight: 59 kDa).Can be blocked with Human Occludin peptide (ab34440). Abcam recommends using 3% milk as the blocking agent.
In-Cell ELISA		Use at an assay dependent concentration. PubMed: 22689949
ICC/IF		Use a concentration of 1 - 2 ?g/ml.
IHC-P		1/100. PubMed: 19828778

Target

FunctionMay play a role in the formation and regulation of the tight junction (TJ) paracellular permeability barrier. It is able to induce adhesion when expressed in cells lacking tight junctions.
Tissue specificityLocalized at tight junctions of both epithelial and endothelial cells. Highly expressed in kidney. Not detected in testis.
Involvement in diseaseDefects in OCLN are the cause of band-like calcification with simplified gyration and polymicrogyria (BLCPMG) [MIM:251290]; also known as pseudo-TORCH syndrome. BLCPMG is a neurologic disorder with characteristic clinical and neuroradiologic features that mimic intrauterine TORCH infection in the absence of evidence of infection. Affected individuals have congenital microcephaly, intracranial calcifications, and severe developmental delay.
Sequence similaritiesBelongs to the ELL/occludin family.
Contains 1 MARVEL domain.
DomainThe C-terminal is cytoplasmic and is important for interaction with ZO-1. Sufficient for the tight junction localization. Involved in the regulation of the permeability barrier function of the tight junction (By similarity). The first extracellular loop participates in an adhesive interaction.
Post-translational
modificationsPhosphorylated upon DNA damage, probably by ATM or ATR. Dephosphorylated by PTPRJ. The tyrosine phosphorylation on Tyr-398 and Tyr-402 reduces its ability to interact with TJP1.
Cellular localizationMembrane. Cell junction > tight junction.
Information by UniProt
Database links
- Entrez Gene: 403844 Dog
- Entrez Gene: 100506658 Human
- Entrez Gene: 397236 Pig
- Entrez Gene: 83497 Rat
- Omim: 602876 Human
- SwissProt: Q28269 Dog
- SwissProt: Q16625 Human
- SwissProt: Q6P6T5 Rat
see all
Alternative names
- BLCPMG antibody
- FLJ08163 antibody
- FLJ18079 antibody
see all

Anti-Occludin antibody images

Western blot - Anti-Occludin antibody (ab31721)
All lanes : Anti-Occludin antibody (ab31721) at 1 ?g/ml

Lane 1 : Colon tissue lysate (Human) Tissue Lysate - adult normal tissue (ab30051)
Lane 2 : Liver (Human) Tissue Lysate - adult normal tissue (ab29889)
Lane 3 : Skeletal Muscle (Human) Tissue Lysate - adult normal tissue (ab29330)

Lysates/proteins at 10 ?g per lane.

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
developed using the ECL technique

Performed under reducing conditions.

Predicted band size : 59 kDa
Observed band size : 63 kDa (why is the actual band size different from the predicted?)
Additional bands at : 240 kDa,32 kDa. We are unsure as to the identity of these extra bands.

Exposure time : 12 minutes
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab31721 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.
Immunocytochemistry/ Immunofluorescence - Occludin antibody (ab31721)
ICC/IF image of ab31721 stained Hek293 cells. The cells were 4% formaldehyde 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 (ab31721, 5?g/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 1.43?M.
Western blot - Anti-Occludin antibody (ab31721)
Anti-Occludin antibody (ab31721) at 1 ?g/ml (blocked with 3% milk) + Kidney (Human) Tissue Lysate - adult normal tissue (ab30203) at 20 ?g

Secondary
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
developed using the ECL technique

Performed under reducing conditions.

Predicted band size : 59 kDa
Observed band size : 63 kDa (why is the actual band size different from the predicted?)
Additional bands at : 26 kDa. We are unsure as to the identity of these extra bands.

Exposure time : 2 minutes

References for Anti-Occludin antibody (ab31721)

This product has been referenced in:

Chung CY et al. Progressive Proximal-to-Distal Reduction in Expression of the Tight Junction Complex in Colonic Epithelium of Virally-Suppressed HIV+ Individuals. PLoS Pathog 10:e1004198 (2014). WB ; Human .Read more (PubMed: 24968145) ?
Sumigray K et al. Cell-cell adhesions and cell contractility are upregulated upon desmosome disruption. PLoS One 9:e101824 (2014). ICC/IF ; Mouse . Read more (PubMed: 25006807) ?

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