Anti-SLFN11 antibody [EPR24414-87] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SLFN11 antibody [EPR24414-87] - BSA and Azide free (AB283232)

This data was developed using 271354, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human colon tissue labelling SLFN11 with 271354 at 1/1000 (0.547 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on stromal cells of human colon. The section was incubated with ab271354 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 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-SLFN11 antibody [EPR24414-87] - BSA and Azide free (AB283232)

This data was developed using 271354, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human colon carcinoma tissue labelling SLFN11 with 271354 at 1/1000 (0.547 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on human colon carcinoma. The section was incubated with ab271354 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-SLFN11 antibody [EPR24414-87] - BSA and Azide free (AB283232)

This data was developed using 271354, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized K-562 (Human chronic myelogenous leukemia lymphoblast, Left) /DU 145 (Human prostate carcinoma epithelial cell, Right) cells labelling SLFN11 with 271354 at 1/500 dilution (0.1ug)/ red (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. Low expression cell line : K-562 (PMID : 29395061).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SLFN11 antibody [EPR24414-87] - BSA and Azide free (AB283232)

This data was developed using 271354, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded A-172 (human brain glioblastoma) (A) and K562 (human chronic myelogenous leukemia lymphoblast) (B) tissue labelling SLFN11 with 271354 at 1/1000 (0.547 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nuclear staining on A-172 cells (A), and no staining on K-562 cells (B). Low expression cell line : K562 (PMID : 29395061).The section was incubated with ab271354 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 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-SLFN11 antibody [EPR24414-87] - BSA and Azide free (AB283232)

This data was developed using 271354, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labelling SLFN11 with 271354 at 1/1000 (0.547 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on human breast cancer. The section was incubated with ab271354 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

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SLFN11 antibody [EPR24414-87] - BSA and Azide free (AB283232)

This data was developed using 271354, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized DU 145 cells labelling SLFN11 with 271354 at 1/50 (10.94 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) Confocl image showing positive staining in DU145 cells. Low expression cell line : K-562 (PMID : 29395061) is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5 ug/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 2ug/ml dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-SLFN11 antibody [EPR24414-87] - BSA and Azide free (AB283232)

This data was developed using 271354, the same antibody clone in a different buffer formulation.

SLFN11 was immunoprecipitated from 0.35 mg DU145 (human prostate carcinoma epithelial cell) whole cell lysate 10 ug with 271354 at 1/30 dilution (2 ug in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using 271354 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : DU145 (human prostate carcinoma epithelial cell) whole cell lysate 10 ug

Lane 2 : ab271354 IP in DU145 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab271354 in DU145 whole cell lysate

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

Exposure time : 5 seconds

All lanes:

Immunoprecipitation - Anti-SLFN11 antibody [EPR24414-87] (<a href='/en-us/products/primary-antibodies/slfn11-antibody-epr24414-87-ab271354'>ab271354</a>)

Predicted band size: 103 kDa

false

• WB

Lab

Western blot - Anti-SLFN11 antibody [EPR24414-87] - BSA and Azide free (AB283232)

This data was developed using ab271354, the same antibody clone in a different buffer formulation.

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

The molecular weight observed is consistent with what has been described in the literature (PMID : 31118262).

Low expression cell line : K562, HEK-293T (PMID : 29395061, 31118262).

Exposure time : 37 seconds

All lanes:

Western blot - Anti-SLFN11 antibody [EPR24414-87] (<a href='/en-us/products/primary-antibodies/slfn11-antibody-epr24414-87-ab271354'>ab271354</a>) at 1/1000 dilution

Lane 1:

DU145 (human prostate carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

A-172 (human brain glioblastoma), whole cell lysate at 20 µg

Lane 3:

K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate at 20 µg

Lane 4:

HEK-293T (human embryonic kidney epithelial cell), whole cell lysate at 20 µg

Lane 5:

Jurkat (human T cell leukemia T lymphocyte), 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: 103 kDa

Observed band size: 75-100 kDa

false

• WB

Lab

Western blot - Anti-SLFN11 antibody [EPR24414-87] - BSA and Azide free (AB283232)

This data was developed using ab271354, the same antibody clone in a different buffer formulation.

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

The molecular weight observed is consistent with what has been described in the literature (PMID : 31118262).

Exposure time : 3 minutes

All lanes:

Western blot - Anti-SLFN11 antibody [EPR24414-87] (<a href='/en-us/products/primary-antibodies/slfn11-antibody-epr24414-87-ab271354'>ab271354</a>) at 1/1000 dilution

Lane 1:

Human lung tissue lysates at 20 µg

Lane 2:

Human lung cancer tissue lysates at 20 µg

Lane 3:

Human colon cancer tissue lysates 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: 103 kDa

Observed band size: 75-100 kDa

false