Anti-CD177 antibody [EPR22813-205]

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD177 antibody [EPR22813-205] (AB255296)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human placenta tissue staining CD68 with ab213363 at a 1/500 dilution, ab315802 anti-CD34 used at 1/2000 dilution and ab255296 anti-CD177 used at a 1/2000 dilution.

Panel A : merged staining of anti-CD68 (green; Opal™520), anti-CD34 (magenta; Opal™690) and anti-CD177 (gray; Opal™570) on human placenta.

Panel B : anti-CD68 staining macrophages in human placenta.

Panel C : ant-CD34 staining endothelium in human placenta.

Panel D : ant-CD177 staining neutrophils in human placenta.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab213363, ab315802 and ab255296 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

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

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD177 antibody [EPR22813-205] (AB255296)

Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling CD177 with ab255296 at 1/2000 dilution (0.252 μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on neutrophils in human spleen (PMID : 24562309) is observed. The section was incubated with ab255296 for 10 mins at RT. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD177 antibody [EPR22813-205] (AB255296)

Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling CD177 with ab255296 at 1/2000 dilution (0.252 μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on neutrophils in human placenta. The section was incubated with ab255296 for 10 mins at RT. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD177 antibody [EPR22813-205] (AB255296)

Immunohistochemical analysis of paraffin-embedded human bone marrow labeling CD177 with ab255296 at 1/2000 dilution (0.252 μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human bone marrow (PMID : 22434870). The section was incubated with ab255296 for 10 mins at RT. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD177 antibody [EPR22813-205] (AB255296)

Immunohistochemistry analysis of Formalin/PFA-fixed paraffin-embedded sections human bone marrow tissue labelling PRG2 with ab322142 at 5000 dilution (B), CD1777 with ab255296 at 2000 dilution (C) and CD42d/GP5 with ab322703 at 500 dilution (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with TrisEDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : anti-PRG2 (green; Opal™520), anti-CD177 (magenta; Opal™690) and anti-CD42d/GP5 (gray; Opal™570) on human bond marrow.

Panel B : anti-PRG2 staining basophils in human bone marrow.

Panel C : anti-CD177 staining neutrophils in human bone marrow.

Panel D : anti-CD42d/PGP5 staining megakaryocytes in human bone marrow.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab322142, ab255296 and ab322703 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD177 antibody [EPR22813-205] (AB255296)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human spleen tissue staining CD42d/GP5 with ab322703 at a 1/500 dilution, ab255296 anti-CD177 used at 1/2000 dilution and ab323609 anti-HYAL2 used at a 1/100 dilution.

Panel A : anti-CD42d/GP5 (green; Opal™520), anti-CD177 (magenta; Opal™690) and anti-HYAL2 (gray; Opal™570) on human spleen.

Panel B : anti-CD42d/GP5 staining platelets in human spleen.

Panel C : anti-CD177 staining neutrophils in human spleen.

Panel D : anti-HYAL2 staining blood vessel in human spleen.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab322703, ab255296 and ab323609 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD177 antibody [EPR22813-205] (AB255296)

Panel A : anti-CD42d/GP5 (green; Opal™520), anti-CD177 (magenta; Opal™690) and anti-HYAL2 (gray; Opal™570) on human spleen. Panel B : anti-CD42d/GP5 staining platelets in human spleen. Panel C : anti-CD177 staining neutrophils in human spleen. Panel D : anti-HYAL2 staining blood vessel in human spleen. Nuclear DNA was labeled with DAPI (shown in blue). The section was incubated in three rounds of staining : in the order of ab322703, ab255296 and ab323609 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD177 antibody [EPR22813-205] (AB255296)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human spleen tissue staining CD42d/GP5 with ab322703 at a 1/500 dilution, ab255296 anti-CD177 used at 1/2000 dilution and ab183322 anti-CD14 used at a 1/100 dilution.

Panel A : anti-CD42d/GP5 (green; Opal™520), anti-CD177 (magenta; Opal™690) and anti-CD14 (gray; Opal™570) on human spleen.

Panel B : anti-CD42d/GP5 staining platelets in human spleen.

Panel C : anti-CD177 staining neutrophils in human spleen.

Panel D : anti-CD14 staining blood vessel in human spleen.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab322703, ab255296 and ab183322 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins.

• WB

Unknown

Western blot - Anti-CD177 antibody [EPR22813-205] (AB255296)

CD177 is a glycoprotein. The molecular weight observed is consistent with what has been described in the literature (PMID : 17580308). Negative control : U-937 (PMID : 17580308).

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-CD177 antibody [EPR22813-205] (ab255296) at 1/1000 dilution

Lane 1:

Human liver lysate at 20 µg

Lane 2:

U-937 (human histiocytic lymphoma monocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 46 kDa

false

Exposure time: 3min

• WB

Unknown

Western blot - Anti-CD177 antibody [EPR22813-205] (AB255296)

CD177 is a glycoprotein. The molecular weight observed is consistent with what has been described in the literature (PMID : 17580308).

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure times : Lane 1 : 26 secs; Lane 2 : 3 mins.

All lanes:

Western blot - Anti-CD177 antibody [EPR22813-205] (ab255296) at 1/1000 dilution

Lane 1:

Human lung lysate at 10 µg

Lane 2:

Human spleen lysate at 10 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 46 kDa

false

• WB

Unknown

Western blot - Anti-CD177 antibody [EPR22813-205] (AB255296)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-CD177 antibody [EPR22813-205] (ab255296) at 1/5000 dilution

All lanes:

Recombinant his-tagged human CD177 protein (aa22-407), 20 ng

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: 46 kDa

false

Exposure time: 59s