Anti-Integrin beta 5 antibody [EPR26930-21]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin beta 5 antibody [EPR26930-21] (AB309092)

Immunohistochemical analysis of paraffin-embedded Human gastric adenoc tissue labeling Integrin beta 5 with ab309092 at 1/500 (1.048 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/Dab (ab209101). Positive staining on human gastric adenocarcinoma (PMID : 32395495). The section was incubated with ab309092 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/Dab (ab209101). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin beta 5 antibody [EPR26930-21] (AB309092)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Integrin beta 5 with ab309092 at 1/500 (1.048 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/Dab (ab209101). Positive staining on human kidney (PMID : 25551756). The section was incubated with ab309092 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/Dab (ab209101). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• Flow Cyt

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Flow Cytometry - Anti-Integrin beta 5 antibody [EPR26930-21] (AB309092)

Flow cytometric analysis of A549 (human lung carcinoma epithelial cell) cells labelling Integrin beta 5 with ab309092 at 1/50 dilution (1 ug)/Red compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody. Gated on viable cells.

• Flow Cyt

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Flow Cytometry - Anti-Integrin beta 5 antibody [EPR26930-21] (AB309092)

Flow cytometric analysis of NIH/3T3 (mouse embryonic fibroblast) cells labelling Integrin beta 5 with ab309092 at 1/50 dilution (1 ug)/Red compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody. Gated on viable cells.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin beta 5 antibody [EPR26930-21] (AB309092)

Immunohistochemical analysis of paraffin-embedded Mouse glioblastoma tissue labeling Integrin beta 5 with ab309092 at 1/500 (1.048 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/Dab (ab209101). Positive staining on mouse glioblastoma. The section was incubated with ab309092 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/Dab (ab209101). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin beta 5 antibody [EPR26930-21] (AB309092)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Integrin beta 5 with ab309092 at 1/500 (1.048 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/Dab (ab209101). Positive staining on mouse kidney. The section was incubated with ab309092 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/Dab (ab209101). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin beta 5 antibody [EPR26930-21] (AB309092)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling Integrin beta 5 with ab309092 at 1/500 (1.048 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/Dab (ab209101). Positive staining on rat kidney. The section was incubated with ab309092 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/Dab (ab209101). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IP

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Immunoprecipitation - Anti-Integrin beta 5 antibody [EPR26930-21] (AB309092)

Integrin beta 5 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate with ab309092 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab309092 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 ug
Lane 2 : ab309092 IP in NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab309092 in NIH/3T3 whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-Integrin beta 5 antibody [EPR26930-21] (ab309092) at 1/1000 dilution

All lanes:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 100 kDa

false

Exposure time: 32s

• WB

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Western blot - Anti-Integrin beta 5 antibody [EPR26930-21] (AB309092)

Blocking and diluting buffer and concentration : 5% NFDM/TBST In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : Lane 1 and 2 : 15 seconds, Lane 3-5 : 10 seconds, Lane 6 and 7 : 103 seconds.

All lanes:

Western blot - Anti-Integrin beta 5 antibody [EPR26930-21] (ab309092) at 1/1000 dilution

Lane 1:

A549 (human lu carcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

A549 transfected with siRNA specifically targeti ITGB5 whole cell lysate at 20 µg

Lane 3:

NCI-H1299 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 5:

Mouse lung tissue lysate at 20 µg

Lane 6:

Mouse liver tissue lysate at 20 µg

Lane 7:

Rat liver tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 100 kDa

false

• Dot

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Dot Blot - Anti-Integrin beta 5 antibody [EPR26930-21] (AB309092)

Dot blot analysis of Integrin beta 5 using ab309092 at 1 : 1000 (0.524 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100000 dilution. Exposure time : 180 seconds Blocking and diluting buffer and concentration : 5% NFDM/TBST