Anti-CXCL10/IP-10 antibody [EPR24674-84]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCL10/IP-10 antibody [EPR24674-84] (AB306587)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling CXCL10/IP-10 with ab306587 at 1/2000 (0.248 µg/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Positive staining on human tonsil (PMID : 12949239).The section was incubated with ab306587 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. 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-CXCL10/IP-10 antibody [EPR24674-84] (AB306587)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling CXCL10/IP-10 with ab306587 at 1/2000 (0.248 µg/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Positive staining on human colonic immune cells (PMID : 10433925; PMID : 24748971).The section was incubated with ab306587 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. 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-CXCL10/IP-10 antibody [EPR24674-84] (AB306587)

Immunohistochemical analysis of paraffin-embedded (A) THP-1 (human mon tissue labeling CXCL10/IP-10 with ab306587 at 1/2000 (0.248 µg/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Positive staining on (A) THP-1 (human monocytic leukemia monocyte) treated with 200ng/ml IFN-y (ab259377) for 24 hours and 50ng/ml LPS for 24 hours, and 5µg/ml Brefeldin A for the last 21 hours cell pellet. No staining on (B) Untreated THP-1 cell pellet. The section was incubated with ab306587 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. 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-CXCL10/IP-10 antibody [EPR24674-84] (AB306587)

CXCL10/IP-10 was immunoprecipitated from 0.35 mg THP-1 treated with 200ng/ml IFN-y (ab259377) for 24 hours and 50ng/ml LPS for 24 hours, and 5μg/ml Brefeldin A for the last 21 hours whole cell lysate 20 μg with ab306587 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab306587. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : THP-1 treated with 200ng/ml IFN-y (ab259377) for 24 hours and 50ng/ml LPS for 24 hours, and 5μg/ml Brefeldin A for the last 21 hours whole cell lysate 20 μg Lane 2 : ab306587 IP in THP-1 treated with 200ng/ml IFN-y (ab259377) for 24 hours and 50ng/ml LPS for 24 hours, and 5μg/ml Brefeldin A for the last 21 hours Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab306587 in THP-1 treated with 200ng/ml IFN-y (ab259377) for 24 hours and 50ng/ml LPS for 24 hours, and 5μg/ml Brefeldin A for the last 21 hours whole cell lysate 20 μg Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 180 seconds

All lanes:

Immunoprecipitation - Anti-CXCL10/IP-10 antibody [EPR24674-84] (ab306587) at 1/30 dilution

All lanes:

THP-1

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: 11 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-CXCL10/IP-10 antibody [EPR24674-84] (AB306587)

Blocking and diluting buffer and concentration : Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS False colour image of Western blot : Anti-CXCL10/IP-10 antibody [EPR24674-84] (ab306587) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab306587 was shown to bind specifically to CXCL10/IP-10. A band was observed at 11 kDa in wild-type A549 cell lysates with no signal observed at this size in CXCL10/IP-10 knockout cell line ab266971 (knockout cell lysate ab256888). To generate this image, wild-type and CXCL10/IP-10 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/10000 dilution.

All lanes:

Western blot - Anti-CXCL10/IP-10 antibody [EPR24674-84] (ab306587) 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 /ml IFN-y (<a href='/en-us/products/proteins-peptides/recombinant-human-interferon-gamma-protein-active-ab259377'>ab259377</a>) for 24 hours and 50ng/ml LPS for 24 hours, and 5ug/ml Brefeldin A for the last 21 hours whole cell lysate at 20 µg

Lane 3:

Untreated Wild-type A549 (human lu carcinoma epithelial cell) whole cell lysate 40 at 40 µg

Lane 4:

Wild-type A549 treated with /ml IFN-y (<a href='/en-us/products/proteins-peptides/recombinant-human-interferon-gamma-protein-active-ab259377'>ab259377</a>) for 32 hours and 10 ng/m TNF-alpha (<a href='/en-us/products/proteins-peptides/recombinant-human-tnf-alpha-protein-active-ab259410'>ab259410</a>) for 32 hours, and 5ug/ml Brefeldin A (<a href='/en-us/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>) for the last 6 hours whole cell lysate 40 at 40 µg

Lane 5:

Untreated CXCL10/IP-10 knockout A549 whole cell lysate at 40 µg

Lane 6:

CXCL10/IP-10 knockout A549 treated with /ml IFN-y (<a href='/en-us/products/proteins-peptides/recombinant-human-interferon-gamma-protein-active-ab259377'>ab259377</a>) for 32 hours and 10 ng/m TNF-alpha (<a href='/en-us/products/proteins-peptides/recombinant-human-tnf-alpha-protein-active-ab259410'>ab259410</a>) for 32 hours, and 5ug/ml Brefeldin A (<a href='/en-us/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>) for the last 6 hours whole cell lys at 40 µg

Secondary

Lanes 1 - 6:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Lanes 1 - 6:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/10000 dilution

Observed band size: 11 kDa

false