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Rabbit Recombinant Monoclonal Occludin antibody. Carrier free. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Rat, Dog, Recombinant full length protein - Human samples. Cited in 2 publications.


Images

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Constituents: PBS

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.
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Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPWBICC/IFFlow Cyt (Intra)IHC-P
Human
Tested
Tested
Tested
Tested
Tested
Mouse
Expected
Tested
Not recommended
Expected
Tested
Rat
Expected
Tested
Not recommended
Expected
Tested
Dog
Expected
Tested
Tested
Expected
Expected
Recombinant full length protein - Human
Not recommended
Tested
Not recommended
Not recommended
Not recommended

Tested
Tested

Species

Human

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species

Mouse, Rat, Dog

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Recombinant full length protein - Human

Dilution info

-

Notes

-

Tested
Tested

Species

Mouse

Dilution info

-

Notes

Please upload more of the lysate or use lower dilution of ab216327 when testing bEnd.3.

Occludin contains multiple isoforms, and the molecular weight range of different isoforms varies greatly (8kDa~59kDa). In addition, there are various post-translational modifications such as ubiquitination and proteolytic cleavage, so the band sizes and number of bands detected in different samples may vary. (PMID: 18647175, PMID: 19821483, PMID: 19457074, PMID: 11782481, PMID: 22083955).

Species

Rat

Dilution info

-

Notes

Please upload more of the lysate or use lower dilution of ab216327 when testing bEnd.3.

Occludin contains multiple isoforms, and the molecular weight range of different isoforms varies greatly (8kDa~59kDa). In addition, there are various post-translational modifications such as ubiquitination and proteolytic cleavage, so the band sizes and number of bands detected in different samples may vary. (PMID: 18647175, PMID: 19821483, PMID: 19457074, PMID: 11782481, PMID: 22083955).

Species

Human

Dilution info

-

Notes

Please upload more of the lysate or use lower dilution of ab216327 when testing bEnd.3.

Occludin contains multiple isoforms, and the molecular weight range of different isoforms varies greatly (8kDa~59kDa). In addition, there are various post-translational modifications such as ubiquitination and proteolytic cleavage, so the band sizes and number of bands detected in different samples may vary. (PMID: 18647175, PMID: 19821483, PMID: 19457074, PMID: 11782481, PMID: 22083955).

Species

Dog

Dilution info

-

Notes

Please upload more of the lysate or use lower dilution of ab216327 when testing bEnd.3.

Occludin contains multiple isoforms, and the molecular weight range of different isoforms varies greatly (8kDa~59kDa). In addition, there are various post-translational modifications such as ubiquitination and proteolytic cleavage, so the band sizes and number of bands detected in different samples may vary. (PMID: 18647175, PMID: 19821483, PMID: 19457074, PMID: 11782481, PMID: 22083955).

Species

Recombinant full length protein - Human

Dilution info

-

Notes

Please upload more of the lysate or use lower dilution of ab216327 when testing bEnd.3.

Occludin contains multiple isoforms, and the molecular weight range of different isoforms varies greatly (8kDa~59kDa). In addition, there are various post-translational modifications such as ubiquitination and proteolytic cleavage, so the band sizes and number of bands detected in different samples may vary. (PMID: 18647175, PMID: 19821483, PMID: 19457074, PMID: 11782481, PMID: 22083955).

Tested
Tested

Species

Dog

Dilution info

-

Notes

This antibody is not suitable for ICC in mouse or rat species.

Species

Human

Dilution info

-

Notes

This antibody is not suitable for ICC in mouse or rat species.

Not recommended
Not recommended

Species

Mouse

Dilution info

-

Notes

This antibody is not suitable for ICC in mouse or rat species.

Species

Rat

Dilution info

-

Notes

This antibody is not suitable for ICC in mouse or rat species.

Species

Recombinant full length protein - Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Expected
Expected

Species

Mouse, Rat, Dog

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Recombinant full length protein - Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species

Mouse

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species

Rat

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species

Dog

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Recombinant full length protein - Human

Dilution info

-

Notes

-

Target data

Function

May 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.(Microbial infection) Acts as a co-receptor for hepatitis C virus (HCV) in hepatocytes.

Alternative names

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Rabbit Recombinant Monoclonal Occludin antibody. Carrier free. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Rat, Dog, Recombinant full length protein - Human samples. Cited in 2 publications.

Alternative names

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free

Yes

Clone number

EPR20992

Purification technique

Affinity purification Protein A

Specificity

This antibody is not suitable for ICC in mouse or rat species or for any testing rat intestinal tissues.

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate long-term storage conditions

+4°C

Storage information

Do Not Freeze

Notes

ab224526 is the carrier-free version of ab216327.

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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 batch production

For more information, read more on recombinant antibodies.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

Supplementary info

Biological function summary

Occludin serves as an important component in maintaining cell-to-cell adhesion within tight junction complexes. It interacts with other tight junction proteins such as claudins and junctional adhesion molecules to regulate paracellular permeability. These interactions help maintain selective barrier properties of epithelial and endothelial layers preventing the passage of large molecules and pathogens while allowing passage of ions and small molecules. Occludin stabilizes the tight junctions by interacting with the cytoskeleton and signaling proteins that are important for tight junction assembly and maintenance.

Activity summary

Occludin is a protein that plays an important mechanical role in forming tight junctions which are specialized connections between neighboring cell membranes. Alternative names for this target include occludin-1. The occludin protein has a molecular weight of approximately 65 kDa. It is expressed in a variety of tissues but levels tend to be high in epithelial and endothelial cells where tight junctions are essential for barrier functions. Occludin acts as integral membrane protein contributing to the complex structures that seal spaces between cells.

Pathways

Occludin functions prominently in the regulation of tight junction integrity and barrier function. It participates in pathways such as the tight junction signaling pathway and the paracellular transport pathway. Occludin interacts with ZO-1 a scaffolding protein bridging its interaction with the actin cytoskeleton which supports the structural organization and function of tight junctions. This connectivity allows for regulation of the permeability and transport across the epithelial barrier.

Associated diseases and disorders

Disruptions in occludin function are connected to conditions such as inflammatory bowel disease (IBD) and some cancers. In IBD abnormal occludin expression can compromise intestinal barrier function leading to increased intestinal permeability and an inflammatory response. In certain cancers loss of occludin expression is linked to increased invasiveness and metastasis due to weakened cell adhesion. The dysfunction of proteins like ZO-1 which interact with occludin in these pathways also contributes to these pathological states highlighting the importance of occludin's regulatory roles in maintaining cellular barrier integrity.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

18 product images

  • Western blot - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526), expandable thumbnail

    Western blot - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526)

    Lanes 1 - 4: Merged signal (red and green). Green - ab216327 observed at 59 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab216327 was shown to recognize Occludin in wild-type HAP1 cells as signal was lost at the expected MW in OCLN (Occludin) knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and OCLN (Occludin) knockout samples were subjected to SDS-PAGE. Ab216327 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution 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.

    Occludin expression in HeLa is expected to be negative.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216327).

    All lanes: Western blot - Anti-Occludin antibody [EPR20992] (AB216327) at 1/1000 dilution

    Lane 1: Wild-type HAP1 whole cell lysate at 40 µg

    Lane 2: OCLN (Occludin) knockout HAP1 whole cell lysate at 40 µg

    Lane 3: HeLa whole cell lysate (Low Occludin expression) at 20 µg

    Lane 4: HepG2 whole cell lysate lysate (High Occludin expression) at 20 µg

    Predicted band size: 59 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526)

    Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Occludin with ab216327 at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Membranous staining on human colon is observed (PMID: 24268521). Counter stained with Hematoxylin.

    Secondary antibody only control: ab209101 (Rabbit specific IHC polymer detection kit HRP/DAB).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216327).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Western blot - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526), expandable thumbnail

    Western blot - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526)

    This data was developed using ab216327, the same antibody clone in a different buffer formulation.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-Occludin antibody [EPR20992] (AB216327) at 1/1000 dilution

    Lane 1: Caco-2 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 2: bEnd.3 (Mouse brain endothelioma) whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051) at 1/10000 dilution

    Predicted band size: 59 kDa

    Observed band size: 65 kDa

    Exposure time: 60s

  • Immunocytochemistry/ Immunofluorescence - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Caco-2 (human colorectal adenocarcinoma cell line) cells labeling Occludin with ab216327 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membrane staining on Caco-2 cell line.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216327).

  • Western blot - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526), expandable thumbnail

    Western blot - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526)

    This data was developed using ab216327, the same antibody clone in a different buffer formulation.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-Occludin antibody [EPR20992] (AB216327) at 1/1000 dilution

    Lane 1: Mouse colon tissue lysate at 20 µg

    Lane 2: Mouse brain tissue lysate at 20 µg

    Lane 3: Mouse lung tissue lysate at 20 µg

    Lane 4: Rat colon tissue lysate at 20 µg

    Lane 5: Rat brain tissue lysate at 20 µg

    Lane 6: Rat lung tissue lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051) at 1/10000 dilution

    Predicted band size: 59 kDa

    Observed band size: 65 kDa

    Exposure time: 100s

  • Immunoprecipitation - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526), expandable thumbnail

    Immunoprecipitation - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526)

    Occludin was immunoprecipitated from 0.35 mg of Caco-2 (human colorectal adenocarcinoma cell line) whole cell lysate with ab216327 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab216327 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: Caco-2 whole cell lysate 10 μg (Input).

    Lane 2: ab216327 IP in Caco-2 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab216327 in Caco-2 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 1 second.

    The molecular weight observed is consistent with what has been described in the literature (PMID: 18647175, PMID: 19821483).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216327).

    All lanes: Immunoprecipitation - Anti-Occludin antibody [EPR20992] (AB216327)

    Predicted band size: 59 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526)

    Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling Occludin with ab216327 at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Cytoplasmic staining on distal tubules of mouse kidney is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: ab209101 (Rabbit specific IHC polymer detection kit HRP/DAB).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216327).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526)

    Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling Occludin with ab216327 at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Cytoplasmic staining on distal tubules of rat kidney is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: ab209101 (Rabbit specific IHC polymer detection kit HRP/DAB).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216327).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216327).

    Immunohistochemical analysis of paraffin-embedded human breast tissue labeling Occludin with ab216327 at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Membranous staining on human breast is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: ab209101 (Rabbit specific IHC polymer detection kit HRP/DAB).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MDCK (canine kidney cell line) cells labeling Occludin with ab216327 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membrane staining on MDCK (NBL-2) cell line.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216327).

  • Flow Cytometry (Intracellular) - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526)

    Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized Caco-2 (human colorectal adenocarcinoma cell line) cell line labeling Occludin with ab216327 at 1/60 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216327).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526)

    Immunohistochemical analysis of paraffin-embedded Human colon labeling Occludin with ab216327 at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Positive staining on human colon. The section was incubated with ab216327 at 4°C overnight. The section was then mounted using Fluoromount®. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    This data was developed using ab216327, the same antibody clone in a different buffer formulation.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216327).
    Tissue Microarrays stained for Anti-Occludin antibody [EPR20992] using ab216327 in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negaive (cross mark) staining per sample type tested. The sections were incubated with ab216327 at 4°C overnight used at 1:2000 dilution (1.05 μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB secondary antibody (ab209101). Counterstain was Hematoxylin.

    Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526)

    Immunohistochemical analysis of paraffin-embedded Mouse colon labeling Occludin with ab216327 at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Positive staining on Mouse colon. The section was incubated with ab216327 at 4°C overnight. The section was then mounted using Fluoromount®. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    This data was developed using ab216327, the same antibody clone in a different buffer formulation.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526)

    Immunohistochemical analysis of paraffin-embedded Rat colon labeling Occludin with ab216327 at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Positive staining on Rat colon. The section was incubated with ab216327 at 4°C overnight. The section was then mounted using Fluoromount®. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    This data was developed using ab216327, the same antibody clone in a different buffer formulation.

  • Western blot - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526), expandable thumbnail

    Western blot - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216327).

    For cells express relatively low level of Occludin, we suggest optimizing experimental protocols (increasing lysate amount, using lower dilution or higher sensitivity ECL substrate) to improve results.

    Blocking and Diluting buffer and concentration: 5% NFDM /TBST

    ab181602 was used as a GAPDH loading control.

    All lanes: Western blot - Anti-Occludin antibody [EPR20992] (AB216327) at 1/1000 dilution

    Lane 1: Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 2: DLD-1(human colorectal adenocarcinoma cell) whole cell lysate at 20 µg

    Lane 3: A431 (human epidermoid carcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 4: A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 5: HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051) at 1/20000 dilution

    Observed band size: 16-60 kDa

    Exposure time: 20s

    For cells express relatively low level of Occludin, we suggest optimizing experimental protocols (increasing lysate amount, using lower dilution or higher sensitivity ECL substrate) to improve results.

    Blocking and Diluting buffer and concentration: 5% NFDM /TBST

    ab181602 was used as a GAPDH loading control.

  • Western blot - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526), expandable thumbnail

    Western blot - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216327).

    Occludin contains multiple isoforms, and the molecular weight range of different isoforms varies greatly (8kDa~59kDa). In addition, there are various post-translational modifications such as ubiquitination and proteolytic cleavage, so the band sizes and number of bands detected in different samples may vary. (PMID: 18647175, PMID: 19821483, PMID: 19457074, PMID: 11782481, PMID: 22083955)

    Blocking and Diluting buffer and concentration: 5% NFDM /TBST

    ab181602 was used as a GAPDH loading control.

    All lanes: Western blot - Anti-Occludin antibody [EPR20992] (AB216327) at 1/1000 dilution

    Lane 1: Mouse colon tissue lysate at 20 µg

    Lane 2: Mouse small intestine tissue lysate at 20 µg

    Lane 3: Mouse ileum tissue lysate at 20 µg

    Lane 4: Mouse liver tissue lysate at 20 µg

    Lane 5: Mouse Kidney tissue lysate at 20 µg

    Lane 6: Human colon tissue lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051) at 1/20000 dilution

    Observed band size: 16-60 kDa

    Exposure time: 100s

    Occludin contains multiple isoforms, and the molecular weight range of different isoforms varies greatly (8kDa~59kDa). In addition, there are various post-translational modifications such as ubiquitination and proteolytic cleavage, so the band sizes and number of bands detected in different samples may vary. (PMID: 18647175, PMID: 19821483, PMID: 19457074, PMID: 11782481, PMID: 22083955)

    Blocking and Diluting buffer and concentration: 5% NFDM /TBST

    ab181602 was used as a GAPDH loading control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Occludin antibody [EPR20992] - BSA and Azide free (ab224526)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216327).

    Immunohistochemical analysis of paraffin-embedded human thyroid carcinoma labeling Occludin with ab216327 at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880), ready to use. Counter stained with Hematoxylin. Antigen retrieval was heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0).

    Secondary antibody only control: ab209101 (Rabbit specific IHC polymer detection kit HRP/DAB).

    Positive staining on human thyroid carcinoma. The section was incubated with ab216327 at 4°C overnight.

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