Anti-CXCL9 antibody [EPR26512-118] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCL9 antibody [EPR26512-118] - BSA and Azide free (AB290654)

This data was developed using ab290643, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labelling CXCL9 with ab290643 at 1/100 (5.38 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Negative control : no staining on human cerebrum. The section was incubated with ab290643 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) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-CXCL9 antibody [EPR26512-118] - BSA and Azide free (AB290654)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized KATO III (Human stomach spherical) cells labelling CXCL9 with primary antibody anti-CXCL9 (ab290643) at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody at 1/1000 dilution. Confocal image showing increased cytoplasmic staining in KATO III cells treated with IFN gamma (human) (100 ng/ml) and TNF alpha (human) (100 ng/ml) for 16 hours. Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) (ab195889) was used to counterstain tubulin at 1/200 dilution. The nuclear counter stain is DAPI (blue). This data was developed using ab290643, the same antibody clone in a different buffer formulation.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCL9 antibody [EPR26512-118] - BSA and Azide free (AB290654)

This data was developed using ab290643, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Panel A THP-1 cell tissue labelling CXCL9 with ab290643 at 1/2000 (0.269 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on (A) THP-1 cells treated with a combination of IFN-γ (200ng/ml, 24h) and LPS (50ng/ml, 24h); no staining on (B) untreated THP-1 cells. The section was incubated with ab290643 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) was used.

Heat mediated antigen retrieval 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-CXCL9 antibody [EPR26512-118] - BSA and Azide free (AB290654)

This data was developed using ab290643, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human prostate carcinoma tissue labelling CXCL9 with ab290643 at 1/100 (5.38 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on human prostate carcinoma. The section was incubated with ab290643 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) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-CXCL9 antibody [EPR26512-118] - BSA and Azide free (AB290654)

This data was developed using ab290643, the same antibody clone in a different buffer formulation.

Intracellular flow cytometric analysis of 4% paraformaldehyde, 90% methanol THP-1 (Human monocytic leukemia monocyte) treated with 200ng/ml IFN gamma and 50ng/ml LPS for 24h (Right) labelling CXCL9 with ab290743 at 1/50 dilution/ Untreated control (Left). Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as a secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCL9 antibody [EPR26512-118] - BSA and Azide free (AB290654)

This data was developed using ab290643, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue labelling CXCL9 with ab290643 at 1/100 dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human lung carcinoma. The section was incubated with ab290643 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) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IP

Supplier Data

Immunoprecipitation - Anti-CXCL9 antibody [EPR26512-118] - BSA and Azide free (AB290654)

This data was developed using ab290643, the same antibody clone in a different buffer formulation.

CXCL9 was immunoprecipitated from 0.35 mg KATO III (human gastric carcinoma) treated with 100ng/ml IFN gamma and 100 ng/ml TNF alpha for 16 hours whole cell lysate 10 ug with ab290643 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab290643 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : KATO III (human gastric carcinoma) treated with 100ng/ml IFN gamma and 100 ng/ml TNF alpha for 16 hours whole cell lysate 10 ug

Lane 2 : ab290643 IP in KATO III treated with 100ng/ml IFN gamma and 100 ng/ml TNF alpha for 16 hours whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab290643 in KATO III treated with 100ng/ml IFN gamma and 100 ng/ml TNF alpha for 16 hours whole cell lysate

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

Exposure time : 3.25 seconds

All lanes:

Immunoprecipitation - Anti-CXCL9 antibody [EPR26512-118] (<a href='/en-us/products/primary-antibodies/cxcl9-antibody-epr26512-118-ab290643'>ab290643</a>)

Predicted band size: 14 kDa

Observed band size: 14 kDa

false

• WB

Supplier Data

Western blot - Anti-CXCL9 antibody [EPR26512-118] - BSA and Azide free (AB290654)

This data was developed using ab290643, the same antibody clone in a different buffer formulation.

Blocking and diluting buffers and concentration was 5% NFDM/TBST.

Exposure time : 7.75 seconds.

The expression of CXCL9 is upregulated in response to TNF gamma and LPS treatment (PMID : 12946268, 20650898).

All lanes:

Western blot - Anti-CXCL9 antibody [EPR26512-118] (<a href='/en-us/products/primary-antibodies/cxcl9-antibody-epr26512-118-ab290643'>ab290643</a>) at 1/1000 dilution

Lane 1:

Untreated THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Lane 2:

THP-1 treated with 200ng/ml IFN gamma and 50 ng/ml LPS(Lipopolysaccharide) 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 1/100000 dilution

Predicted band size: 14 kDa

Observed band size: 14 kDa

false

• WB

Supplier Data

Western blot - Anti-CXCL9 antibody [EPR26512-118] - BSA and Azide free (AB290654)

This data was developed using ab290643, the same antibody clone in a different buffer formulation.

Blocking and diluting buffers and concentration was 5% NFDM/TBST.

Exposure time : 5.5 seonds.

The expression of CXCL9 is upregulated in response to IFN gamma and TNF alpha treatment (PMID : 12946268, 20650898).

All lanes:

Western blot - Anti-CXCL9 antibody [EPR26512-118] (<a href='/en-us/products/primary-antibodies/cxcl9-antibody-epr26512-118-ab290643'>ab290643</a>) at 1/1000 dilution

Lane 1:

Untreated KATO III (human gastric carcinoma) whole cell lysate at 20 µg

Lane 2:

KATO III treated with 100ng/ml IFN gamma and 100 ng/ml TNF alpha for 16 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 1/100000 dilution

Predicted band size: 14 kDa

Observed band size: 14 kDa

false

• WB

Supplier Data

Western blot - Anti-CXCL9 antibody [EPR26512-118] - BSA and Azide free (AB290654)

This data was developed using ab290643, the same antibody clone in a different buffer formulation.

Blocking and diluting buffers and concentration was 5% NFDM/TBST.

Exposure time : 10 seonds.

This antibody does not cross-react with CXCL10, CXCL11, or CXCL2 protein.

All lanes:

Western blot - Anti-CXCL9 antibody [EPR26512-118] (<a href='/en-us/products/primary-antibodies/cxcl9-antibody-epr26512-118-ab290643'>ab290643</a>) at 1/1000 dilution

Lane 1:

HEK-293T (human embryonic kidney) cells transfected with a human CXCL9 expression vector containing a myc-His tag whole cell lysate at 4 µg

Lane 2:

HEK-293T cells transfected with a human CXCL10 expression vector containing a myc-His tag whole cell lysate at 20 µg

Lane 3:

HEK-293T cells transfected with a human CXCL11 expression vector containing a myc-His tag whole cell lysate at 40 µg

Lane 4:

HEK-293T cells transfected with a human CXCL2 expression vector containing a myc-His tag 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 1/100000 dilution

Predicted band size: 14 kDa

Observed band size: 14 kDa

false

Exposure time: 10s