Anti-Fragilis antibody [EPR26405-14] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fragilis antibody [EPR26405-14] - BSA and Azide free (AB288569)

This data was developed using ab288563, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat lung tissue labelling Interferon-induced transmembrane protein 3/IFITM3 with ab288563 at 1/500 (1.088 ug/ml) followed by a ready to use LeicaDS9800 (BOND^®, Polymer Refine Detection) was used. Positive staining on rat lung. The section was incubated with ab288563 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 Leica DS9800 (BOND^®, Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-Fragilis antibody [EPR26405-14] - BSA and Azide free (AB288569)

This data was developed using ab288563, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cerebrum (fresh) tissue labeling Interferon-induced transmembrane protein 3/IFITM3 with ab288563 at 1/500 (1.088 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green). Positive staining on mouse cerebrum is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbedat 1/1000 (2 ug/mL) dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Fragilis antibody [EPR26405-14] - BSA and Azide free (AB288569)

This data was developed using ab288563, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 cells labelling Interferon-induced transmembrane protein 3/IFITM3 with ab288563 at 1/2000 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in NIH/3T3 cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 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 dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fragilis antibody [EPR26405-14] - BSA and Azide free (AB288569)

This data was developed using ab288563, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labelling Interferon-induced transmembrane protein 3/IFITM3 with ab288563 at 1/500 (1.088 ug/ml) followed by a ready to use Leica DS9800 (BOND^®, Polymer Refine Detection) was used. Positive staining on endothelium of mouse cerebrum. The section was incubated with ab288563 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 Leica DS9800 (BOND^®, Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fragilis antibody [EPR26405-14] - BSA and Azide free (AB288569)

This data was developed using ab288563, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labelling Interferon-induced transmembrane protein 3/IFITM3 with ab288563 at 1/500 (1.088 ug/ml) followed by a ready to use Leica DS9800 (BOND^®, Polymer Refine Detection) was used. Positive staining on mouse lung. The section was incubated with ab288563 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 Leica DS9800 (BOND^®, Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-Fragilis antibody [EPR26405-14] - BSA and Azide free (AB288569)

This data was developed using ab288563, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse lung (fresh) tissue labeling Interferon-induced transmembrane protein 3/IFITM3 with ab288563 at 1/500 (1.088 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green). Positive staining on mouse lung is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbedat 1/1000 (2 ug/mL) dilution.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-Fragilis antibody [EPR26405-14] - BSA and Azide free (AB288569)

This data was developed using ab288563, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat lung (fresh) tissue labeling Interferon-induced transmembrane protein 3/IFITM3 with ab288563 at 1/500 (1.088 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green). Positive staining on rat lung is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbedat 1/1000 (2 ug/mL) dilution.

• IP

Lab

Immunoprecipitation - Anti-Fragilis antibody [EPR26405-14] - BSA and Azide free (AB288569)

This data was developed using ab288563, the same antibody clone in a different buffer formulation.

Interferon-induced transmembrane protein 3/IFITM3 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 ug with ab288563 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab288563 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 ug

Lane 2 : ab288563 IP in NIH/3T3 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab288563 in NIH/3T3 whole cell lysate

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

Exposure time : 3.25 seconds

All lanes:

Immunoprecipitation - Anti-Fragilis antibody [EPR26405-14] (<a href='/en-us/products/primary-antibodies/fragilis-antibody-epr26405-14-ab288563'>ab288563</a>)

Predicted band size: 15 kDa

false

• WB

Lab

Western blot - Anti-Fragilis antibody [EPR26405-14] - BSA and Azide free (AB288569)

This data was developed using ab288563, the same antibody clone in a different buffer formulation.

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

Samples are non-boiled as boiling may cause protein aggregates.

Exposure time : 15 seconds

All lanes:

Western blot - Anti-Fragilis antibody [EPR26405-14] (<a href='/en-us/products/primary-antibodies/fragilis-antibody-epr26405-14-ab288563'>ab288563</a>) at 1/1000 dilution

Lane 1:

Untreated NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 20 µg

Lane 2:

NIH/3T3 treated with 100ng/ml IFN gamma for 48 hours, whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 15 kDa

false

• WB

Lab

Western blot - Anti-Fragilis antibody [EPR26405-14] - BSA and Azide free (AB288569)

This data was developed using ab288563, the same antibody clone in a different buffer formulation.

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

Samples are non-boiled as boiling may cause protein aggregates.

This blot was developed using a higher sensitivity ECL substrate.

Exposure time : Lane1-2 : 15 seconds; Lane3-4 : 3 minutes;

All lanes:

Western blot - Anti-Fragilis antibody [EPR26405-14] (<a href='/en-us/products/primary-antibodies/fragilis-antibody-epr26405-14-ab288563'>ab288563</a>) at 1/1000 dilution

Lane 1:

Mouse lung tissue lysate at 20 µg

Lane 2:

Mouse liver tissue lysate at 20 µg

Lane 3:

Mouse kidney tissue lysate at 20 µg

Lane 4:

Rat lung tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 15 kDa

false