Occludin antibody (AA 431-522)

Catalog No. ABIN687337

This independently validated Rabbit Polyclonal antibody specifically detects Occludin in WB, ELISA, FACS, IF (cc), IF (p) and IHC (p). It exhibits reactivity toward Human, Mouse and Pig and has been mentioned in 5+ publications.

Quick Overview for Occludin antibody (AA 431-522) (ABIN687337)

Target: Occludin (OCLN)

Reactivity: Human, Mouse, Pig

Host: Rabbit

Clonality: Polyclonal

Conjugate: This Occludin antibody is un-conjugated

Application: Western Blotting (WB), ELISA, Flow Cytometry (FACS), Immunofluorescence (Cultured Cells) (IF (cc)), Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))

Product Details anti-Occludin Antibody

Binding Specificity: AA 431-522

Cross-Reactivity: Human, Mouse, Pig

Predicted Reactivity: Rat,Dog,Cow,Horse,Guinea Pig

Purification: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from mouse Occludin

Isotype: IgG

Application Details

Application Notes: WB 1:300-5000
ELISA 1:500-1000
FCM 1:20-100
IHC-P 1:200-400
IF(IHC-P) 1:50-200
IF(ICC) 1:50-200

Restrictions: For Research Use only

Independent Validation (IF)

Validation #029632 (Immunofluorescence)

by: Confocal Imaging Core, Beth Israel Deaconess Medical Center

No.: #029632

Date: 03/23/2014

Lot Number: 990698W

Method validated: Immunofluorescence

Positive Control: Mouse stomach tissue

Negative Control: Human fat tissue

Notes: Signal was detected in positive control sample and not in negative control sample.

Validation Images

Figure 1: Immunostaining (green) of mouse stomach tissue (A-C) and human fat tissue (D-F) with anti-OCLN (A, D), isotype control antibody (B, E), or with secondary alone (C, F). Tissues were counterstained with DAPI (blue).

Full Methods

Primary Antibody:

• Antibody: Occludin (OCLN)
• Catalog number: ABIN687337
• Lot number: 990698W

Secondary Antibody:

• Antibody: Donkey anti-Rabbit IgG (Heavy & Light Chain) Antibody (Alexa 647)

Full Protocol:

• Paraffin embedded positive and negative control tissue sections were deparaffinized and undergone antigen retrieval using 10 mM Sodium Citrate pH6 in a pressure cooker for 10 min.
• The tissue sections were incubated with 1 mg/mL Sodium borohydride for 10 min at room temperature to block autofluorescent background signal. The sections were then rinsed three times in TBS for 5 min each at RT.
• Tissue sections were blocked in 1 X TBS / 5% normal donkey serum for 60 min at RT.
• Tissue sections were incubated with primary antibody diluted 1:200 in 1X PBS / 5% normal donkey serum overnight at 4°C.
• Tissue sections were rinsed three times in TBS for 5 min each at RT.
• Cells were incubated with secondary antibody diluted 1:200 in 1X PBS / 5% normal donkey serum for 60 min at RT in dark.
• Tissue sections were rinsed three times in TBS for 5 min each at RT.
• Coverslips were mounted on slides with Prolong Gold anti-fade mounting media (Invitrogen)
• IF stained tissue sections were imaged with a Zeiss LSM 510 Meta confocal microscope.

Experimental Notes:

• No challenges notes
• OCLN is recommended to staining of mouse stomach tissue according to this protocol.

Handling

Format: Liquid

Concentration: 1 μg/μL

Buffer: 0.01M TBS( pH 7.4) with 1 % BSA, 0.02 % Proclin300 and 50 % Glycerol.

Preservative: ProClin

Precaution of Use: This product contains ProClin: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.

Storage: 4 °C,-20 °C

Storage Comment: Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.

Expiry Date: 12 months

References for anti-Occludin antibody (ABIN687337)

Sun, Liu, Xue, Wang: "Effects of insulin combined with idebenone on blood-brain barrier permeability in diabetic rats." in: Journal of neuroscience research, Vol. 93, Issue 4, pp. 666-77, (2015) (PubMed).

Zhao, Qin, Han, Wang, Zhang, Liu: "?-Conglycinin reduces the tight junction occludin and ZO-1 expression in IPEC-J2." in: International journal of molecular sciences, Vol. 15, Issue 2, pp. 1915-26, (2014) (PubMed).

Pan, Qin, Zhao, Wang, Liu, Che: "Effects of soybean agglutinin on mechanical barrier function and tight junction protein expression in intestinal epithelial cells from piglets." in: International journal of molecular sciences, Vol. 14, Issue 11, pp. 21689-704, (2013) (PubMed).

Zhao, Qin, Sun, Che, Bao, Zhang: "Effects of soybean agglutinin on intestinal barrier permeability and tight junction protein expression in weaned piglets." in: International journal of molecular sciences, Vol. 12, Issue 12, pp. 8502-12, (2012) (PubMed).

Gu, Xue, Wei, Zhang, Li: "Calcium-activated potassium channel activator down-regulated the expression of tight junction protein in brain tumor model in rats." in: Neuroscience letters, Vol. 493, Issue 3, pp. 140-4, (2011) (PubMed).

Target Details for Occludin

Target: Occludin (OCLN)

Alternative Name: Occludin

Background:

Synonyms: Ocl, AI53564, Occludin, Ocln

Background: May play a role in the formation and regulation of the tight junction (TJ) paracellular permeability barrier.

Gene ID: 18260

UniProt: Q61146

Pathways: Cell-Cell Junction Organization, Hepatitis C