Anti-CD9 antibody [EPR27551-92] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD9 antibody [EPR27551-92] - BSA and Azide free (AB307086)

This data was developed using ab307085, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling CD9 with ab307085 at 1/2000 (0.228 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Membranous staining in rat kidney. The section was incubated with ab307085 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-CD9 antibody [EPR27551-92] - BSA and Azide free (AB307086)

This data was developed using ab307085, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling CD9 with ab307085 at 1/2000 (0.228 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining in mouse kidney. The section was incubated with ab307085 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-CD9 antibody [EPR27551-92] - BSA and Azide free (AB307086)

This data was developed using ab307085, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling CD9 with ab307085 at 1/2000 (0.228 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining in mouse cerebrum. The section was incubated with ab307085 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-CD9 antibody [EPR27551-92] - BSA and Azide free (AB307086)

This data was developed using ab307085, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Mouse lung cancer tissue labeling CD9 with ab307085 at 1/2000 (0.228 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Membranous staining in mouse lung cancer. The section was incubated with ab307085 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-CD9 antibody [EPR27551-92] - BSA and Azide free (AB307086)

This data was developed using ab307085, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling CD9 with ab307085 at 1/2000 (0.228 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining in rat cerebrum. The section was incubated with ab307085 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-CD9 antibody [EPR27551-92] - BSA and Azide free (AB307086)

This data was developed using ab307085, the same antibody clone in a different buffer formulation. CD9 was immunoprecipitated from 0.35 mg RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate 10 μg with ab307085 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307085 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate 10 μg Lane 2 : ab307085 IP in RAW 264.7 whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab307085 in RAW 264.7 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 10 seconds

All lanes:

Immunoprecipitation - Anti-CD9 antibody [EPR27551-92] (<a href='/en-us/products/primary-antibodies/cd9-antibody-epr27551-92-ab307085'>ab307085</a>) at 1/30 dilution

All lanes:

RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage)

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: 23 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-CD9 antibody [EPR27551-92] - BSA and Azide free (AB307086)

This data was developed using ab307085, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS The samples were run on a Bis-Tris gel. Performed under reducing conditions. False colour image of Western blot : Anti-CD9 antibody [EPR27551-92] (ab307085) 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, ab307085 was shown to bind specifically to CD9. A band was observed at 23 kDa in wild-type HeLa cell lysates whereas no signal observed at this size in CD9 knockout cell line ab255375 (knockout cell lysate ab263754). To generate this image, wild-type and CD9 knockout HeLa cell lysates were analyzed. First, samples were run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in in Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS 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 1/20000 dilution.

All lanes:

Western blot - Anti-CD9 antibody [EPR27551-92] (<a href='/en-us/products/primary-antibodies/cd9-antibody-epr27551-92-ab307085'>ab307085</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

CD9 knockout HeLa whole cell lysate at 20 µg

Lane 3:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Secondary

Lanes 1 - 4:

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 - 4:

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: 23 kDa

false

• WB

Supplier Data

Western blot - Anti-CD9 antibody [EPR27551-92] - BSA and Azide free (AB307086)

This data was developed using ab307085, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative controls : K-562, Raji (PMID : 17407154 ; PMID : 8921952). Exposure time : 70 seconds

All lanes:

Western blot - Anti-CD9 antibody [EPR27551-92] (<a href='/en-us/products/primary-antibodies/cd9-antibody-epr27551-92-ab307085'>ab307085</a>) at 1/1000 dilution

Lane 1:

HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Lane 3:

Raji (human Burkitts lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 4:

Mouse brain tissue lysate at 20 µg

Lane 5:

Mouse kidney tissue lysate at 20 µg

Lane 6:

C6 (rat glial tumor glial cell) 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

Observed band size: 23 kDa

false

Exposure time: 70s