CD11b Antibody [F15D4]

Catalog No.: F0040

Application: Reactivity: Mouse

当該製品は品切れ状态で、ごメールアドレスを教えていただければ、在庫があると、メールで顧客様に伝えます。

代表番号: 045-509-1970|電子メール：sales@selleck.co.jp

キーポイント

この抗体には抗ラット二次抗体が必要です。

使用情報

Dilution
IHC1:100 FCM1:4000

Application
IP, IHC, FCM

Source
Rat Monoclonal Antibody

Reactivity
Mouse

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW
165-170 kDa

ポジティブコントロール	Mouse bone marrow; Mouse spleen tissue
ネガティブコントロール

プロトコール

IHC
Experimental Protocol： Deparaffinization/Rehydration 1. Deparaffinize/hydrate sections: 2. Incubate sections in three washes of xylene for 5 min each. 3. Incubate sections in two washes of 100% ethanol for 10 min each. 4. Incubate sections in two washes of 95% ethanol for 10 min each. 5. Wash sections two times in dH2O for 5 min each. 6.Antigen retrieval: For Citrate: Heat slides in a microwave submersed in 1X citrate unmasking solution until boiling is initiated; continue with 10 min at a sub-boiling temperature (95°-98°C). Cool slides on bench top for 30 min. Staining 1. Wash sections in dH2O three times for 5 min each. 2. Incubate sections in 3% hydrogen peroxide for 10 min. 3. Wash sections in dH2O two times for 5 min each. 4. Wash sections in wash buffer for 5 min. 5. Block each section with 100–400 µl of blocking solution for 1 hr at room temperature. 6. Remove blocking solution and add 100–400 µl primary antibody diluent in to each section. Incubate overnight at 4°C. 7. Remove antibody solution and wash sections with wash buffer three times for 5 min each. 8. Cover section with 1–3 drops HRPas needed. Incubate in a humidified chamber for 30 min at room temperature. 9. Wash sections three times with wash buffer for 5 min each. 10. Add DAB Chromogen Concentrate to DAB Diluent and mix well before use. 11. Apply 100–400 µl DAB to each section and monitor closely. 1–10 min generally provides an acceptable staining intensity. 12. Immerse slides in dH2O. 13. If desired, counterstain sections with hematoxylin. 14. Wash sections in dH2O two times for 5 min each. 15. Dehydrate sections: Incubate sections in 95% ethanol two times for 10 sec each; Repeat in 100% ethanol, incubating sections two times for 10 sec each; Repeat in xylene, incubating sections two times for 10 sec each. 16. Mount sections with coverslips and mounting medium.

IHC

Experimental Protocol：

Deparaffinization/Rehydration

1. Deparaffinize/hydrate sections:

2. Incubate sections in three washes of xylene for 5 min each.

3. Incubate sections in two washes of 100% ethanol for 10 min each.

4. Incubate sections in two washes of 95% ethanol for 10 min each.

5. Wash sections two times in dH2O for 5 min each.

6.Antigen retrieval: For Citrate: Heat slides in a microwave submersed in 1X citrate unmasking solution until boiling is initiated; continue with 10 min at a sub-boiling temperature (95°-98°C). Cool slides on bench top for 30 min.

Staining

1. Wash sections in dH2O three times for 5 min each.

2. Incubate sections in 3% hydrogen peroxide for 10 min.

3. Wash sections in dH2O two times for 5 min each.

4. Wash sections in wash buffer for 5 min.

5. Block each section with 100–400 µl of blocking solution for 1 hr at room temperature.

6. Remove blocking solution and add 100–400 µl primary antibody diluent in to each section. Incubate overnight at 4°C.

7. Remove antibody solution and wash sections with wash buffer three times for 5 min each.

8. Cover section with 1–3 drops HRPas needed. Incubate in a humidified chamber for 30 min at room temperature.

9. Wash sections three times with wash buffer for 5 min each.

10. Add DAB Chromogen Concentrate to DAB Diluent and mix well before use.

11. Apply 100–400 µl DAB to each section and monitor closely. 1–10 min generally provides an acceptable staining intensity.

12. Immerse slides in dH2O.

13. If desired, counterstain sections with hematoxylin.

14. Wash sections in dH2O two times for 5 min each.

15. Dehydrate sections: Incubate sections in 95% ethanol two times for 10 sec each; Repeat in 100% ethanol, incubating sections two times for 10 sec each; Repeat in xylene, incubating sections two times for 10 sec each.

16. Mount sections with coverslips and mounting medium.

Datasheet & SDS

生物学的記述

Specificity
CD11b Antibody [F15D4] detects endogenous levels of total CD11b protein.

タンパク質の局在
細胞質、細胞核

Uniprot ID
P21127

Clone
F15D4

Synonym(s)
CD11 antigen-like family member B; CD11B (p170); cell surface glycoprotein MAC-1 alpha subunit; cell surface glycoprotein MAC-1 subunit alpha; complement component receptor 3 alpha-a; Mac1A; Cd11b; CD11b/CD18; CR3; CR3A; F730045J24Rik; Ly-40; Mac-1; Mac-1a; MAC1

Background
CD11b, also known as integrin alpha M (ITGAM), is a transmembrane alpha chain protein that non-covalently heterodimerizes with the beta-2 chain CD18 to form the Mac-1 (αMβ2 or CR3) integrin, which is prominently expressed on myeloid cells such as neutrophils, monocytes, macrophages, and dendritic cells, serving as a key marker for these innate immune populations. This integrin contains a β-propeller domain and a ligand-binding inserted (I/A) domain in its alpha subunit, allowing interactions with diverse ligands including iC3b, ICAM-1, and fibrinogen. Disease-associated ITGAM variants such as R77H (rs1143679), P1146S (rs1143678), and A858V (rs1143683) subtly alter the β-propeller structure, reducing ligand affinity without significantly affecting surface expression. CD11b mediates proinflammatory processes like leukocyte adhesion, migration, transmigration, and phagocytosis of opsonized particles (apoptotic cells and immune complexes), while also exerting important anti-inflammatory effects by negatively regulating Toll-like receptor (TLR) signaling, via Src/Syk-mediated phosphorylation and Cbl-b-dependent ubiquitination and degradation of MyD88 and TRIF, to suppress NF-κB activation and proinflammatory cytokine production (IL-6, TNF-α, IL-1β). CD11b also curtails type I interferon (IFN-I) production through an AKT/FOXO3/IRF3/7 pathway, maintaining nuclear FOXO3 to repress IRF7 and IFN-β/IRF7 expression; this mechanism is defective in ITGAM variant carriers, leading to elevated basal IFN-I, uncontrolled TLR responses, impaired B-cell tolerance, enhanced autoreactive B-cell activation, and increased Th17 differentiation. These immune dysregulations confer a high genetic risk for systemic lupus erythematosus (SLE) and lupus nephritis (LN), with ITGAM variants being associated with higher IFN-I serum levels, renal involvement, autoantibody production, glomerular injury, and poor immune complex clearance.

Background

CD11b, also known as integrin alpha M (ITGAM), is a transmembrane alpha chain protein that non-covalently heterodimerizes with the beta-2 chain CD18 to form the Mac-1 (αMβ2 or CR3) integrin, which is prominently expressed on myeloid cells such as neutrophils, monocytes, macrophages, and dendritic cells, serving as a key marker for these innate immune populations. This integrin contains a β-propeller domain and a ligand-binding inserted (I/A) domain in its alpha subunit, allowing interactions with diverse ligands including iC3b, ICAM-1, and fibrinogen. Disease-associated ITGAM variants such as R77H (rs1143679), P1146S (rs1143678), and A858V (rs1143683) subtly alter the β-propeller structure, reducing ligand affinity without significantly affecting surface expression. CD11b mediates proinflammatory processes like leukocyte adhesion, migration, transmigration, and phagocytosis of opsonized particles (apoptotic cells and immune complexes), while also exerting important anti-inflammatory effects by negatively regulating Toll-like receptor (TLR) signaling, via Src/Syk-mediated phosphorylation and Cbl-b-dependent ubiquitination and degradation of MyD88 and TRIF, to suppress NF-κB activation and proinflammatory cytokine production (IL-6, TNF-α, IL-1β). CD11b also curtails type I interferon (IFN-I) production through an AKT/FOXO3/IRF3/7 pathway, maintaining nuclear FOXO3 to repress IRF7 and IFN-β/IRF7 expression; this mechanism is defective in ITGAM variant carriers, leading to elevated basal IFN-I, uncontrolled TLR responses, impaired B-cell tolerance, enhanced autoreactive B-cell activation, and increased Th17 differentiation. These immune dysregulations confer a high genetic risk for systemic lupus erythematosus (SLE) and lupus nephritis (LN), with ITGAM variants being associated with higher IFN-I serum levels, renal involvement, autoantibody production, glomerular injury, and poor immune complex clearance.

References
[1] Schmid MC, et al. Nat Commun. 2018 Dec 19;9(1):5379. [2] Fagerholm SC, et al. Blood. 2006 Nov 15;108(10):3379-86.

PVDFメンブレン孔径参考表

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

技術サポート

ストックの作り方、阻害剤の保管方法、細胞実験や動物実験の際に注意すべき点など、製品を取扱う時に問い合わせが多かった質問に対しては取扱説明書でお答えしています。

Handling Instructions

他に質問がある場合は、お気軽にお問い合わせください。

* 必須

* 大学・企業名 大学・企業名を記入してください

* 名前 名前を記入してください

* 電子メール 電子メール・アドレスを記入してください 有効なメールアドレスを入力してください

電話番号

* 内容 お問い合わせ内容をご入力ください

Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%