Anti-S100A8 + S100A9 antibody [RM1038]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100A8 + S100A9 antibody [RM1038] (AB288715)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling S100A8+S100A9 with ab288715 at 1/5000 (0.096 ug/ml) followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Positive staining on the human spleen. The section was incubated with ab288715 for 10 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 Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-S100A8 + S100A9 antibody [RM1038] (AB288715)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HL-60 cells labelling S100A8+S100A9 with ab288715 at 1/500 (0.96 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). Confocal image showing membranous staining in HL-60 cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 (2.5ug/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 (2 ug/ml) dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100A8 + S100A9 antibody [RM1038] (AB288715)

Immunohistochemical analysis of paraffin-embedded Human stomach carcin tissue labeling S100A8+S100A9 with ab288715 at 1/5000 (0.096 ug/ml) followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Positive staining on the stroma inflammatory cells in human stomach carcinoma. The section was incubated with ab288715 for 10 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 Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-S100A8 + S100A9 antibody [RM1038] (AB288715)

Flow cytometric analysis of 4% paraformaldehyde fixed 0.1% Tween-20 permeabilized HL-60 (Human acute promyelocytic leukemia promyeloblast) cells labelling S100A8+S100A9 with ab288715 at 1/500 dilution (0.1ug)(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, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IP

Lab

Immunoprecipitation - Anti-S100A8 + S100A9 antibody [RM1038] (AB288715)

S100A8+S100A9 was immunoprecipitated from 0.35 mg HL-60 (Human acute promyelocytic leukemia promyeloblast) whole cell lysate 10ug with ab288715 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab288715 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HL-60 (Human acute promyelocytic leukemia promyeloblast) whole cell lysate 10ug

Lane 2 : ab288715 IP in HL-60 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab288715 in HL-60 whole cell lysate

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

Exposure time : 30 seconds

All lanes:

Immunoprecipitation - Anti-S100A8 + S100A9 antibody [RM1038] (ab288715)

Predicted band size: 13 kDa

Observed band size: 11 kDa,14 kDa

false

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-S100A8 + S100A9 antibody [RM1038] (AB288715)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Mouse PBMC cells labelling S100A8+S100A9 with ab288715 at 1/500 (0.96 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). Confocal image showing membranous staining in subsets of mouse PBMCs is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 (2.5ug/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 (2 ug/ml) dilution.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-S100A8 + S100A9 antibody [RM1038] (AB288715)

Flow cytometric analysis of 4% paraformaldehyde fixed 0.1% Tween-20 permeabilized Mouse blood cells cells labelling S100A8+S100A9 with ab288715 at 1/500 dilution (0.1 μg)/ Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control. A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100A8 + S100A9 antibody [RM1038] (AB288715)

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling S100A8+S100A9 with ab288715 at 1/5000 (0.096 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on the rat spleen. The section was incubated with ab288715 for 10 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 Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100A8 + S100A9 antibody [RM1038] (AB288715)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling S100A8+S100A9 with ab288715 at 1/5000 (0.096 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on the mouse spleen. The section was incubated with ab288715 for 10 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 Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-S100A8 + S100A9 antibody [RM1038] (AB288715)

Positive staining on the red pulp of mouse spleen.

Fresh mouse spleen was fixed with 4% PFA and parmeabilised with 0.2 % Triton X100. ab288715 was used as a primary antibody at 1/100 dilution. ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) was preadsorbed and used as a sacondary antiobdy at 1/1000 dilution. DAPI was used as a nuclear counter stain.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-S100A8 + S100A9 antibody [RM1038] (AB288715)

Positive staining on the red pulp of rat spleen.

Fresh rat spleen was fixed with 4% PFA and parmeabilised with 0.2 % Triton X100. ab288715 was used as a primary antibody at 1/100 dilution. ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) was preadsorbed and used as a sacondary antiobdy at 1/1000 dilution. DAPI was used as a nuclear counter stain.

• IP

Lab

Immunoprecipitation - Anti-S100A8 + S100A9 antibody [RM1038] (AB288715)

S100A8+S100A9 was immunoprecipitated from 0.35 mg Mouse spleen lysate 10ug with ab288715 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab288715 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse spleen lysate 10ug

Lane 2 : ab288715 IP in Mouse spleen lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab288715 in Mouse spleen lysate

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

Exposure time : 10 seconds

All lanes:

Immunoprecipitation - Anti-S100A8 + S100A9 antibody [RM1038] (ab288715)

Predicted band size: 13 kDa

Observed band size: 11 kDa,14 kDa

false

• WB

Lab

Western blot - Anti-S100A8 + S100A9 antibody [RM1038] (AB288715)

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

Exposure time : 3 min

All lanes:

Western blot - Anti-S100A8 + S100A9 antibody [RM1038] (ab288715) at 1/1000 dilution

Lane 1:

Mouse spleen lysate

Lane 2:

Rat spleen lysate

Lane 3:

SK-BR-3 (Human breast adenocarcinoma epithelial cell) whole cell lysate

Lane 4:

HL-60 (Human acute promyelocytic leukemia promyeloblast) whole cell lysate

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: 13 kDa

Observed band size: 11 kDa,14 kDa

false

• WB

Lab

Western blot - Anti-S100A8 + S100A9 antibody [RM1038] (AB288715)

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

Exposure time : 3 min

All lanes:

Western blot - Anti-S100A8 + S100A9 antibody [RM1038] (ab288715) at 1/1000 dilution

Lane 1:

Mouse PBMC lysate

Lane 2:

Mouse spleen lysate

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: 13 kDa

Observed band size: 11 kDa,14 kDa

false