Anti-TRIM9 antibody [EPR25103-71]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM9 antibody [EPR25103-71] (AB300515)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling TRIM9 with ab300515 at 1/5000 (0.095 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Negative control : no staining on mouse liver (PMID : 20085810). The section was incubated with ab300515 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

• WB

Supplier Data

Western blot - Anti-TRIM9 antibody [EPR25103-71] (AB300515)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Negative control : human liver tissue (PMID : 20085810).

The antibody detects isoform of TRIM9 around 61kDa which consistent with what has been described in the literature (PMID : 20085810).

The identity of the lower MW band at approximately 37 kDa (in lane 1, 2) is unknown.

The blot of lane 5 was developed using a high sensitivity ECL substrate.

Exposure time : Lane 1-4 : 136 seconds Lane 5 : 3 minutes

All lanes:

Western blot - Anti-TRIM9 antibody [EPR25103-71] (ab300515) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 20 µg

Lane 2:

Human liver tissue lysate at 40 µg

Lane 3:

IMR-32 (human neuroblastoma neuroblast), whole cell lysate at 20 µg

Lane 4:

SK-N-BE(2) (human neuroblastoma neuroblast), whole cell lysate at 20 µg

Lane 5:

Y79 (human retinoblastoma retinoblastoma), 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: 79 kDa

Observed band size: 61 kDa,79 kDa

false

• WB

Supplier Data

Western blot - Anti-TRIM9 antibody [EPR25103-71] (AB300515)

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

The identity of the bands between 37 kDa and 25 kDa are unknown.

Exposure time : Lane 1-5 : 81 seconds Lane 6, 7 : 52 minutes

All lanes:

Western blot - Anti-TRIM9 antibody [EPR25103-71] (ab300515) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse hippocampus tissue lysate at 20 µg

Lane 3:

Mouse liver tissue lysate at 40 µg

Lane 4:

Mouse cerebellum tissue lysate at 20 µg

Lane 5:

Rat cerebellum tissue lysate at 20 µg

Lane 6:

Rat brain tissue lysate at 20 µg

Lane 7:

Rat liver tissue lysate at 40 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: 79 kDa

Observed band size: 61 kDa,79 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM9 antibody [EPR25103-71] (AB300515)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling TRIM9 with ab300515 at 1/5000 (0.095 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Negative control : no staining on mouse spleen (PMID : 20085810). The section was incubated with ab300515 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-TRIM9 antibody [EPR25103-71] (AB300515)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling TRIM9 with ab300515 at 1/5000 (0.095 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on mouse cerebrum (PMID : 20085810). The section was incubated with ab300515 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-TRIM9 antibody [EPR25103-71] (AB300515)

TRIM9 was immunoprecipitated from 0.35 mg Mouse brain tissue lysate 10 ug with ab300515 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300515 at ID188 dilution. VeriBlot for IP secondary antibody (HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : Mouse brain tissue lysate 10 ug Lane 2 : ab300515 IP in Mouse brain tissue lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab300515 in mouse brain tissue lysate. Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 41 seconds

Lane 1:

Immunoprecipitation - Anti-TRIM9 antibody [EPR25103-71] (ab300515) at 1/30 dilution

Lane 2:

Immunoprecipitation - Anti-TRIM9 antibody [EPR25103-71] (ab300515) at 1/1000 dilution

All lanes:

Mouse brain tissue lysate 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

false

Exposure time: 41s

• IP

Supplier Data

Immunoprecipitation - Anti-TRIM9 antibody [EPR25103-71] (AB300515)

TRIM9 was immunoprecipitated from 0.35 mg SK-N-BE(2) (human neuroblastoma neuroblast), whole cell lysate 10 ug with ab300515 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300515 at ID188 dilution. VeriBlot for IP secondary antibody (HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : SK-N-BE(2) (human neuroblastoma neuroblast), whole cell lysate 10 ug Lane 2 : ab300515 IP in SK-N-BE(2) whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab300515 in SK-N-BE(2) whole cell lysate. Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 23 seconds

Lane 1:

Immunoprecipitation - Anti-TRIM9 antibody [EPR25103-71] (ab300515) at 1/30 dilution

Lane 2:

Immunoprecipitation - Anti-TRIM9 antibody [EPR25103-71] (ab300515) at 1/1000 dilution

All lanes:

SK-N-BE(2) whole cell lysate 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

false

Exposure time: 23s