Anti-Histamine antibody [EPR28884-78]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histamine antibody [EPR28884-78] (AB324589)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Histamine with ab324589 at 1/10000 (0.051 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mast cells in human colon. The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histamine antibody [EPR28884-78] (AB324589)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Histamine with ab324589 at 1/10000 (0.051 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on human kidney. The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histamine antibody [EPR28884-78] (AB324589)

Immunohistochemical analysis of paraffin-embedded Human lung carcinoma tissue labeling Histamine with ab324589 at 1/10000 (0.051 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mast cells in human lung carcinoma. The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• mIHC

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Multiplex immunohistochemistry - Anti-Histamine antibody [EPR28884-78] (AB324589)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human colon tissue staining Histamine with ab324589 at a 1 : 10000 (0.051 ug/ml) dilution, ab283653 anti-c-Kit used at 1 : 100 (5.73 ug/ml) dilution.

Panel A : merged staining of anti-Histamine (green; Opal™520) and anti-c-Kit (magenta; Opal™690) on human colon.
Panel B : anti-Histamine staining mast cells in human colon.
Panel C : ant-c-Kit staining mast cells in human colon.
Panel D : Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324589 and ab283653 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.

Nuclear counter stain with DAPI.

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

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-Histamine antibody [EPR28884-78] (AB324589)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse stomach (fresh frozen) tissue labeling Histamine with ab324589 at 1/50 (10.16 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Confocal image showing positive staining on mouse stomach. The nuclear counterstain was DAPI (Blue). The section was incubated with the primary antibody for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histamine antibody [EPR28884-78] (AB324589)

Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling Histamine with ab324589 at 1/500 (1.016 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endocrine cells of rat colon. The primary antibody was incubated 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 was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histamine antibody [EPR28884-78] (AB324589)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling Histamine with ab324589 at 1/500 (1.016 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endocrine cells of mouse colon. The primary antibody was incubated 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 was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• cELISA

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Competitive ELISA - Anti-Histamine antibody [EPR28884-78] (AB324589)

0.1 µg/mL Histamine-crosslink-BSA was coated onto a 96-well plate followed by addition of a serial dilution of free Histamine, 3-Methylhistamine dihydrochloride, 4-(2-aminoethyl)-phenol, Histidine, L-Phenylalanine, Tryptamine or Serotonin, as indicated, (0-1000 µg/mL). 0.05 µg/mL ab324589 (50 µL) was then added into each well, before the plate was incubated for 30 minutes and washed. 1 : 20000 diluted GAR Fc-HRP was added and incubated for 30 minutes before washing again. TMB (100 µL) was added and incubated for 5-10 minutes without shaking for color development. Finally, 100 µL of stop solution was added and the OD read at 450 nm on a microplate reader within 5-10 minutes.

• I-ELISA

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Indirect ELISA - Anti-Histamine antibody [EPR28884-78] (AB324589)

Indirect ELISA analysis of ab324589 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1 : 2500 dilution dilution.

Antigen : Histamine-crosslink-BSA.

Antigen concentration : 1000 ng/ml

• cELISA

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Competitive ELISA - Anti-Histamine antibody [EPR28884-78] (AB324589)

ab324589 (used at 0.05 µg/mL) selectively binds to Histamine, with minimal cross-reactivity to other compounds, as shown in the table.