CD11b Recombinant Rabbit Monoclonal Antibody [JU93-81]
cat.: ET1706-04
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, IP, IHC-P, IF-Cell, IF-Tissue, mIHC
Clonality: Monoclonal
Clone number: JU93-81
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 127 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human CD11b 1103-1152 / 1152.
Positive control: TF-1 cell lysate, THP-1 cell lysate, U-937 cell lysate, U937, THP-1, TF-1, human tonsil tissue, human lymph nodes tissue, human spleen tissue, human gastric cancer.
Subcellular location: Cell membrane, Membrane raft.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  IP
  mIHC
1:1,000
1:50-1:200
1:50-1:200
1:200-1:1,000
Use at an assay dependent concentration.
1:1,000
Uniprot #: SwissProt: P11215 Human
Alternative names: antigen CD11b (p170) Antigen CD11b p170 CD11 antigen like family member B CD11 antigen-like family member B CD11b CD11b/CD18 CD49d Cell surface glycoprotein MAC-1 subunit alpha Complement component 3 receptor 3 subunit Complement Component Receptor 3 Alpha Complement receptor type 3, alpha subunit CR 3 alpha chain (CR3A) CR 3 alpha chain CR-3 alpha chain CR3 CR3A F730045J24Rik Integrin Alpha M Integrin alpha M chain Integrin alpha-M Integrin beta 2 alpha subunit Integrin subunit alpha M integrin, alpha M (complement component 3 receptor 3 subunit) ITAM_HUMAN ITGAM Leukocyte adhesion receptor MO1 Ly-40 MAC 1 Mac-1a MAC1 Mac1, alpha subunit MAC1A Macrophage antigen alpha polypeptide MGC117044 Mo1, alpha subunit MO1A Neutrophil adherence receptor alpha M subunit Neutrophil adherence receptor SLEB6
Images
ET1706-04_1.jpg Fig1: Western blot analysis of CD11b on different lysates with Rabbit anti-CD11b antibody (ET1706-04) at 1/1,000 dilution.

Lane 1: TF-1 cell lysate (10 µg/Lane)
Lane 2: THP-1 cell lysate (15 µg/Lane)
Lane 3: U-937 cell lysate (30 µg/Lane)
Lane 4: Jurkat cell lysate (negative) (10 µg/Lane)

Predicted band size: 127 kDa
Observed band size: 170 kDa

Exposure time: 1 minute 50 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1706-04) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1706-04_2.jpg Fig2: Fluorescence multiplex immunohistochemical analysis of the human gastric cancer (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-CD31 (M1511-8, red), anti-αSMA (ET1607-53, gray), anti-CD11b (ET1706-04, cyan), anti-panCK (HA601138, magenta) and anti-CD3 (HA720082, yellow) on human gastric cancer. Panel B: anti- CD31 stained on the endothelial cells. Panel C: anti-αSMA stained on cancer-associated fibroblasts and smooth muscle cells. Panel D: anti-CD11b stained on myeloid cells. Panel E: anti-panCK stained on cancer cells. Panel F: anti-CD3 stained on T cells. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKit™MH010101, www.luminiris.cn). The section was incubated in five rounds of staining: in the order of M1511-8 (1/1,000 dilution), ET1607-53 (1/2,000 dilution), ET1706-04 (1/1,000 dilution), HA601138 (1/3,000 dilution), and HA720082 (1/500 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.
ET1706-04_3.jpg Fig3: Fluorescence multiplex immunohistochemical analysis of human gastric cancer (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-CD11b (ET1706-04, Red), anti-CD3 (HA720082, Green) and anti-CD31 (M1511-8, Yellow) on human gastric cancer. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKit™MH010101, www.luminiris.cn). The section was incubated in three rounds of staining: in the order of ET1706-04 (1/1,000 dilution), HA720082 (1/500 dilution) and M1511-8 (1/1,000 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Zeiss Observer 7 Inverted Fluorescence Microscope.
ET1706-04_4.jpg Fig4: Fluorescence multiplex immunohistochemical analysis of human cervical carcinoma (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-CD66b (HA500100, Green), anti-CD11b (ET1706-04, Red) and anti-CD68 (EM1901-95, Yellow) on human cervical carcinoma. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKit™MH010101, www.luminiris.cn). The section was incubated in three rounds of staining: in the order of HA500100 (1/1,000 dilution), ET1706-04 (1/1,000 dilution) and EM1901-95 (1/3,000 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Zeiss Observer 7 Inverted Fluorescence Microscope.
ET1706-04_5.jpg Fig5: Immunofluorescence analysis of paraffin-embedded human lymph nodes tissue labelling CD11 b (ET1706-04).

The human lymph node section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes, blocked in 10% goat serum, and then incubated with ET1706-04 at 1/50 dilution , followed by iFluorTM 488 Conjugated Goat anti-rabbit IgG at 1:1000 dilution. Nuclear was stained with Hoechst 33258 at 1/5,000 dilution.
Confocal images shows specific membrane staining of CD11b in human lymph node.
ET1706-04_6.jpg Fig6: Immunofluorescence analysis of paraffin-embedded human spleen tissue labelling CD11 b (ET1706-04).

The human spleen section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes, blocked in 10% goat serum, and then incubated with ET1706-04 at 1/50 dilution , followed by iFluorTM 488 Conjugated Goat anti-rabbit IgG at 1:1000 dilution. Nuclear was stained with Hoechst 33258 at 1/5,000 dilution.
Confocal images shows specific membrane staining of CD11b in human spleen.
ET1706-04_7.jpg Fig7: Immunocytochemistry analysis of U937 cells labeling CD11b with Rabbit anti-CD11b antibody (ET1706-04) at 1/50 dilution.

Cells were fixed in 100% methanol for 10 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 2% normal goat serum for 1 hour at 37℃. Cells were then incubated with Rabbit anti-CD11b antibody (ET1706-04) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) were used as the secondary antibody at 1/1,000 dilution.
ET1706-04_8.jpg Fig8: Immunocytochemistry analysis of TF-1 cells labeling CD11b with Rabbit anti-CD11b antibody (ET1706-04) at 1/50 dilution.

Cells were fixed in 100% methanol for 10 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 2% normal goat serum for 1 hour at 37℃. Cells were then incubated with Rabbit anti-CD11b antibody (ET1706-04) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) were used as the secondary antibody at 1/1,000 dilution.
ET1706-04_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-CD11b antibody (ET1706-04) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1706-04) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1706-04_10.jpg Fig10: Immunohistochemical analysis of paraffin-embedded human lymph nodes tissue with Rabbit anti-CD11b antibody (ET1706-04) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1706-04) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1706-04_11.jpg Fig11: Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-CD11b antibody (ET1706-04) at 1/800 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1706-04) at 1/800 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1706-04_12.jpg Fig12: Immunocytochemistry analysis of THP-1(+) Jurkat(-) cells labeling CD11b with Rabbit anti-CD11b antibody (ET1706-04) at 1/100 dilution.

Cells were fixed in 100% precooled methanol for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-CD11b antibody (ET1706-04) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
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