Anti-S100A12/CGRP antibody [EPR23677-111]

• mIHC

Lab

Multiplex immunohistochemistry - Anti-S100A12/CGRP antibody [EPR23677-111] (AB272713)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human Hodgkin's lymphoma tissue staining CD15 with ab135377 at a 1/50 dilution and ab272713 anti-S100A12/CGRP used at 1/5000 dilution. Opal Polymer HRP Ms + Rb was used as a secondary, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-CD15 (green; Opal^™520) and anti-S100A12/CGRP (magenta; Opal^™690) on human Hodgkin's lymphoma.
Panel B : anti-CD15 stained granulocytes and monocytes on human Hodgkin's lymphoma.
Panel C : anti-S100A12/CGRP stained neutrophils and monocytes on human Hodgkin's lymphoma.
Panel D : Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab135377 and ab272713 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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100A12/CGRP antibody [EPR23677-111] (AB272713)

Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling S100A12/CGRP with ab272713 at 1/5000 dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on granulocytes of human spleen (PMID : 10464253). The section was incubated with ab272713 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-S100A12/CGRP antibody [EPR23677-111] (AB272713)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human spleen tissue staining CD15 with ab135377 at a 1/50 dilution and ab272713 anti-S100A12/CGRP used at 1/5000 dilution. Opal Polymer HRP Ms + Rb was used as a secondary, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-CD15 (green; Opal^™520), anti-S100A12/CGRP (magenta; Opal^™690) on human spleen.
Panel B : anti-CD15 stained granulocytes and monocytes on human spleen.
Panel C : anti-S100A12/CGRP stained neutrophils and monocytes on human spleen.
Panel D : Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab135377 and ab272713 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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100A12/CGRP antibody [EPR23677-111] (AB272713)

Immunohistochemical analysis of paraffin-embedded human cervical cancer tissue labeling S100A12/CGRP with ab272713 at 1/5000 dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on granulocytes of human cervical cancer (PMID : 31993369). The section was incubated with ab272713 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 ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-S100A12/CGRP antibody [EPR23677-111] (AB272713)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized human PBMC cells labelling S100A12/CGRP with ab272713 at 1/100 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 2 μg/ml dilution (Green). Confocal image showing cytoplasmic staining in subsets of human PBMC cell line. 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 ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 2 μg/ml dilution.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-S100A12/CGRP antibody [EPR23677-111] (AB272713)

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed, 0.1% Tween-20 permeabilized human peripheral blood mononuclear cell (PBMC) cells labeling S100A12/CGRP with ab272713 at 1/400 dilution (0.1μg) (Right panel), compared with a Rabbit monoclonal IgG (ab172730) isotype control (Left panel). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100A12/CGRP antibody [EPR23677-111] (AB272713)

Immunohistochemical analysis of paraffin-embedded human bladder cancer tissue labeling S100A12/CGRP with ab272713 at 1/5000 dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on granulocytes of human bladder cancer (PMID : 31993369). The section was incubated with ab272713 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 ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IP

Unknown

Immunoprecipitation - Anti-S100A12/CGRP antibody [EPR23677-111] (AB272713)

S100A12/CGRP was immunoprecipitated from 0.35 mg human lymph node tissue lysate 10 μg with ab272713 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab272713 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Human lymph node tissue lysate 10 μg.

Lane 2 : ab272713 IP in Human lymph node tissue lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab272713 in human lymph node tissue lysate.

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

Exposure time : 50 seconds.

All lanes:

Immunoprecipitation - Anti-S100A12/CGRP antibody [EPR23677-111] (ab272713)

Predicted band size: 10 kDa

false

• WB

Lab

Western blot - Anti-S100A12/CGRP antibody [EPR23677-111] (AB272713)

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

The molecular weight observed is consistent with what has been described in the literature (PMID : 19542470).

Negative control : Human brain (PMID : 8718672).

Exposure time : 103 seconds.

All lanes:

Western blot - Anti-S100A12/CGRP antibody [EPR23677-111] (ab272713) at 1/1000 dilution

Lane 1:

Human spleen tissue lysate at 40 µg

Lane 2:

Human lymph node tissue lysate at 40 µg

Lane 3:

Human brain tissue lysate at 40 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 1/50000 dilution

Predicted band size: 10 kDa

Observed band size: 10 kDa

false