CD11b Recombinant Rabbit Monoclonal Antibody [JU93-81]

☑ Relative expression (RE)

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: 3 minutes; ECL: K1801;

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.

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.

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.

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.

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.

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.

☑ Relative expression (RE)

Immunocytochemistry analysis of THP-1 (positive) and Jurkat (negative) 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.

Immunocytochemistry analysis of U-937 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.

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.

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.

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.

☑ Cell treatment (CT)

Western blot analysis of CD11b on different lysates with Rabbit anti-CD11b antibody (ET1706-04) at 1/2,000 dilution.

Lane 1: HL-60 (Human acute promyelocytic leukemia cell) cell lysate
Lane 2: HL-60 treated with 1.3% DSMO for 5 days cell lysate

Lysates/proteins at 20 µg/Lane.
Exposure time: 1 minute 16 seconds; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: ET1706-04, 1/2,000 in 5% NFDM/TBST, overnight at 4 ℃
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 127 kDa
Observed band size: 200 kDa