Anti-CD11b antibody [EPR1344]

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD11b antibody [EPR1344] (AB133357)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse spleen tissue staining LY6C with ab317272 at a 1 : 100 (0.978 ug/ml) dilution, ab133357 anti-CD11b used at 1 : 20000 (0.05 ug/ml) dilution and ab245235 anti-CD19 used at a 1 : 1000 (0.999 ug/ml) dilution.

Panel A : merged staining of anti-Ly6c (green; Opal™690), anti-CD11b (magenta; Opal™570) and anti-CD19 (yellow; Opal™520) on mouse spleen.
Panel B : anti-Ly6c stained on monocytes/macrophages.
Panel C : anti-CD11b stained on monocytes/macrophages.
Panel D : anti-CD19 stained on B cells.
Co-staining of Ly6c and CD11b can be observed.
The section was incubated in three rounds of staining : in the order of ab317272, ab133357, and ab245235 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.
Nuclear DNA was labeled with DAPI (shown in blue).

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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD11b antibody [EPR1344] (AB133357)

Immunohistochemical staining of paraffin embedded human spleen with purified ab133357 at a 1/4000 dilution. The secondary antibody used is a HRP goat anti-rabbit (ab97051). The sample is counterstained with hematoxylin.

Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.

PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD11b antibody [EPR1344] (AB133357)

Immunohistochemical analysis of formalin fixed paraffin embedded human spleen labelling CD11b with ab133357 at a concentration of 0.32 µg/ml. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with a Bond™ Polymer Refine Detection kit. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins. ab133357Anti-CD11b antibody was incubated for 30 mins at room temperature. Sections were counterstained with Hematoxylin. Image inset shows absence of staining in secondary antibody only control.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD11b antibody [EPR1344] (AB133357)

IHC image of CD11b staining in a formalin fixed, paraffin embedded human normal spleen tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab133357 at 1/4000 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD11b antibody [EPR1344] (AB133357)

Immunohistochemical analysis of formalin fixed paraffin embedded human spleen labelling CD11b with ab133357 at a concentration of 1µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32 mins at 100°C with ULTRA cell conditioning solution (CC1, pH 8.5). ab133357 Anti-CD11b antibody was incubated at 37°C for 16 mins. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD11b antibody [EPR1344] (AB133357)

Tissue Microarrays stained for "Anti-CD11b antibody [EPR1344]" using "ab133357"in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. The sections were incubated with ab133357 for 30 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• IHC-P

PubMed

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD11b antibody [EPR1344] (AB133357)

Rottlerin decreases the number of effector cells that mainly infiltrate the skin in IMQ-treated mice

Immunohistochemical detection of immune cell-related markers was performed on paraffin-embedded sections obtained from the back skin of IMQ-induced mice treated with vehicle or rottlerin.

Representatives IHC images of CD11b (B) on the skin of the vehicle or rottlerin-treated mice. Scale bar = 100μm.

Quantification analysis of IHC staining for CD11b(E) on the skin of the vehicle and rottlerin treated mice. Two independent researchers counted the number of positive staining cells were per high-power field (HPF). The data are representative of three experiments (n = 5 mice per group). ** P<0.01 vs. vehicle.

Min, M et al PLoS One. 2017 Dec 22;12(12):e0190051. doi: 10.1371/journal.pone.0190051. eCollection 2017 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

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Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD11b antibody [EPR1344] (AB133357)

Tissue Microarrays stained for "Anti-CD11b antibody [EPR1344]" using "ab133357"in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. The sections were incubated with ab133357 for 30 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• IHC-P

AbReview63941****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD11b antibody [EPR1344] (AB133357)

Formalin-fixed, 0.2% Triton-X permeabilized Mouse tissue sections (Eo771 breast cancer) was stained for CD11b using ab133357 (Green) at 1/1000 dilution in immunohistochemical analysis. The secondary antibody was a Alexa Fluor^® 488 Goat anti-Rabbit IgG (H+L) at 1/1000 dilution.

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration : 4% · Temperature : 25°C

This image is courtesy of an abreview from Anugraha Rajagopalan

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD11b antibody [EPR1344] (AB133357)

ab133357 staining CD11b in paraffin embedded Rat cerebrum tissue sections by Immunohistochemistry. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Samples were incubated with primary antibody at 1/4000 dilution (0.29 μg/ml). A ready to use Goat Anti-rabbit IgG H&L (HRP) was used as the secondary antibody. Hematoxylin was used as a counterstain. Positive staining on gliocytes of rat cerebrum [PMID : 20483006].

• IHC-P

AbReview53642****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD11b antibody [EPR1344] (AB133357)

Formaldehyde-fixed, paraffin-embedded rat bone marrow tissue stained for CD11b using ab133357 at 1/5000 in immunohistochemical analysis.

Heat mediated antigen retrieval with EDTA buffer pH 9 was performed before commencing with staining protocol. 1% casein was used as blocking agent.

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Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD11b antibody [EPR1344] (AB133357)

ab133357 staining CD11b in paraffin embedded Mouse colon tissue sections by Immunohistochemistry. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Samples were incubated with primary antibody at 1/4000 dilution (0.031 μg/ml). A ready to use Goat Anti-rabbit IgG H&L (HRP) was used as the secondary antibody. Hematoxylin was used as a counterstain. Positive staining on stromal cells of mouse colon.

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Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD11b antibody [EPR1344] (AB133357)

ab133357 staining CD11b in paraffin embedded Mouse lung tissue sections by Immunohistochemistry. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Samples were incubated with primary antibody at 1/4000 dilution (0.031 μg/ml). A ready to use Goat Anti-rabbit IgG H&L (HRP) was used as the secondary antibody. Hematoxylin was used as a counterstain. Positive staining on stromal cells of mouse lung.

• WB

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Western blot - Anti-CD11b antibody [EPR1344] (AB133357)

Blocking buffer : 5% NFDM/TBST

Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-CD11b antibody [EPR1344] (ab133357) at 1/1000 dilution

Lane 1:

TF-1 cell lysate at 20 µg

Lane 2:

TPA treated THP-1 cell lysate at 20 µg

Secondary

All lanes:

HRP goat anti-rabbit (H+L) at 1/1000 dilution

Predicted band size: 127 kDa

Observed band size: 170 kDa

false

• WB

Lab

Western blot - Anti-CD11b antibody [EPR1344] (AB133357)

Blocking buffer : 5% NFDM/TBST

Dilution buffer : 5% NFDM/TBST

All lanes:

purified at 1/10000 dilution

Lane 1:

RAW264.7 cell lysate at 20 µg

Lane 2:

Mouse spleen tissue lysate at 20 µg

Secondary

All lanes:

HRP goat anti-rabbit (H+L) at 1/1000 dilution

Predicted band size: 127 kDa

Observed band size: 170 kDa

false

• WB

Lab

Western blot - Anti-CD11b antibody [EPR1344] (AB133357)

Blocking and diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-CD11b antibody [EPR1344] (ab133357) at 1/1000 dilution

All lanes:

Rat spleen tissue lysate at 20 µg

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 1/20000 dilution

Predicted band size: 127 kDa

Observed band size: 170 kDa

false

Exposure time: 8s

• WB

Lab

Western blot - Anti-CD11b antibody [EPR1344] (AB133357)

Different batches of ab133357 were tested on Rat spleen lysate at 0.1 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 170 kDa.

All lanes:

Western blot - Anti-CD11b antibody [EPR1344] (ab133357)

Predicted band size: 127 kDa

false