Anti-Periostin antibody [RM1074]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Periostin antibody [RM1074] (AB315104)

Immunohistochemical analysis of paraffin-embedded Human lung cancer tissue labeling Periostin with ab315104 at 1/1000 (0.511 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive extracellular matrix staining on human lung cancer.
The section was incubated with ab315104 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Periostin antibody [RM1074] (AB315104)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SK-N-SH (human neuroblastoma epithelial cell) cells labelling Periostin with ab315104 at 1/100 (5.11 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing cytoplasmic staining in SK-N-SH cell line.
Negative control : LNCap.
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/500 (1ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Periostin antibody [RM1074] (AB315104)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Periostin with ab315104 at 1/1000 (0.511 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive extracellular matrix staining on human colon.
The section was incubated with ab315104 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Periostin antibody [RM1074] (AB315104)

Immunohistochemical analysis of paraffin-embedded Human breast tissue labeling Periostin with ab315104 at 1/1000 (0.511 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive extracellular matrix staining on human breast.
The section was incubated with ab315104 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IP

Supplier Data

Immunoprecipitation - Anti-Periostin antibody [RM1074] (AB315104)

Periostin was immunoprecipitated from 0.35 mg SK-N-SH (Human neuroblastoma) whole cell lysate with ab315104 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab315104 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : SK-N-SH (Human neuroblastoma) whole cell lysate

Lane 2 : ab315104 at 1/30 IP in SK-N-SH (Human neuroblastoma) whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab315104 in SK-N-SH whole cell lysate

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Immunoprecipitation - Anti-Periostin antibody [RM1074] (ab315104) at 1/1000 dilution

Lane 1:

SK-N-SH (Human neuroblastoma) whole cell lysate at 10 µg

Lane 2:

ab315104 at 1/30 IP in SK-N-SH at 10 µg

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

true

Exposure time: 47s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Periostin antibody [RM1074] (AB315104)

Immunohistochemical analysis of paraffin-embedded Rat stomach tissue labeling Periostin with ab315104 at 1/1000 (0.511 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive extracellular matrix staining on rat stomach.
The section was incubated with ab315104 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Periostin antibody [RM1074] (AB315104)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat stomach (fresh) tissue labeling Periostin with ab315104 at 1/50 (10.22 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green).

Confocal image showing positive staining on rat stomach. The nuclear counterstain was DAPI (Blue). The section was incubated with ab315104 for 60 mins at room temperature. The section was then mounted using Fluoromount®. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 (2 ug/mL) dilution.

• WB

Supplier Data

Western blot - Anti-Periostin antibody [RM1074] (AB315104)

Negative control : Human skeletal muscle (PMID : 15731169).

The expression profile observed in human skeletal muscle is consistent with what has been described in the literature (PMID : 15731169).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Periostin antibody [RM1074] (ab315104) at 1/1000 dilution

Lane 1:

Human breast tissue lysate at 20 µg

Lane 2:

Human skeletal muscle tissue lysate at 20 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/100000 dilution

Observed band size: 95 kDa,36 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Anti-Periostin antibody [RM1074] (AB315104)

Periostin is a secreted protein and Brefeldin A which could inhibit protein transport from the endoplasmic reticulum to the golgi complex increases the detected signal of Periostin in cell lysate.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Periostin antibody [RM1074] (ab315104) at 1/1000 dilution

Lane 1:

Untreated SK-N-SH (Human neuroblastoma) whole cell lysate at 20 µg

Lane 2:

SK-N-SH treated with /ml Brefeldin A for 24 hours whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 95 kDa,36 kDa

false

Exposure time: 6s

• I-ELISA

Supplier Data

Indirect ELISA - Anti-Periostin antibody [RM1074] (AB315104)

Indirect ELISA analysis of ab315104 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1 : 2500 dilution.

Antigen : Mouse Periostin.

Antigen concentration : 1000 ng/ml

• I-ELISA

Supplier Data

Indirect ELISA - Anti-Periostin antibody [RM1074] (AB315104)

Indirect ELISA analysis of ab315104 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1 : 2500 dilution.

Antigen : Human Periostin.

Antigen concentration : 1000 ng/ml