Anti-CD14 抗体 [SP192]

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD14 antibody [SP192] (AB183322)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human tonsil tissue staining C3a R with ab317321 at a 1 : 1000 (0.52 ug/ml) dilution, ab183322 anti-CD14 used at 1 : 100 (1.22 ug/ml) dilution and ab16669 anti-CD3 epsilon used at a 1 : 150 (0.06 ug/ml) dilution.

Panel A : merged staining of anti-C3AR1 (green; Opal™520) anti-CD14 (red; Opal™570) anti-CD3 epsilon (magenta; Opal™690) on human tonsil.
Panel B : anti-C3AR1 stained on immune cells.
Panel C : anti-CD14 stained on macrophages.
Panel D : anti-CD3 epsilon stained on T lymphocytes.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab317321, ab183322 and ab16669 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 was performed with Tris-EDTA buffer (pH 9.0 Epitope Retrieval Solution2) for 20 mins.

• Flow Cyt

Lab

Flow Cytometry - Anti-CD14 antibody [SP192] (AB183322)

Flow cytometry staining of human peripheral blood mononuclear cells (PBMCs) with ab183322 (right) or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (left). PBMCs were incubated for 30 mins on ice in 1x PBS containing 10 µg/ml human IgG and 10 % normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab183322 or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (1x 10⁶ in 100 µl at 0.008 μg/ml) for 30 mins on ice. The cells were simultaneously stained with CD19.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed was incubated at 1/4000 dilutionfor 30 mins on ice

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on CD19-ve.

• Flow Cyt

Lab

Flow Cytometry - Anti-CD14 antibody [SP192] (AB183322)

Flow cytometry staining of human peripheral blood mononuclear cells (PBMCs) with ab183322 (right) or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (left). PBMCs were incubated for 30 min on ice in 1x PBS containing 10 µg/ml human IgG and 10 % normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab183322 or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (1x 10⁶ in 100 µl at 0.008 μg/ml) for 30min on ice. The cells were simultaneously stained with CD19.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min on ice

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on live cells.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [SP192] (AB183322)

The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Paraffin embedded human tonsil tissue slides were dewaxed, followed by heat mediated antigen retrieval with Citrate buffer (pH 6.0, Epitope Retrieval ER1 Solution) for 10 mins. Endogenous peroxidase activity was blocked with 3% H2O2 for 10 mins. The sections were then incubated with ab183322 (1/100) at room temperature for 30 mins, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). The sections were counterstained with Hematoxylin. The images were taken with a Leica Aperio AT2.

ab183322 was stored at a range of temperatures (+4°C, +22°C, +37°C) for 1 week before being tested in IHC-P. The image shows the staining intensity remains relatively constant across all storage temperatures, demonstrating that antibody activity is not affected.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD14 antibody [SP192] (AB183322)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human liver tissue staining TLR2 with ab325499 at a 1 : 100 (5.0 µg/ml) dilution, ab183322 anti-CD14 used at 1 : 100 (1.22 µg/ml) dilution and ab245118 anti-CD8 alpha used at a 1 : 500 (0.75 µg/ml) dilution.

Panel A : anti-TLR2 (green; Opal™520), anti-CD14 (magenta; Opal™570) and anti-CD8 alpha (yellow; Opal™690) on human liver.
Panel B : anti-TLPR2 staining immune cells in human liver.
Panel C : anti-CD14 staining macrophages in human liver.
Panel D : anti-CD8 alpha staining T lymphocytes in human liver.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab325499, ab183322 and ab245118 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [SP192] (AB183322)

Immunohistochemical analysis of paraffin embedded Human tonsil tissue labeling CD14 with ab183322 at 1/100.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [SP192] (AB183322)

Immunohistochemical analysis of paraffin embedded Human esophagus tissue labeling CD14 with ab183322 at 1/100.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [SP192] (AB183322)

Immunohistochemical analysis of paraffin embedded Human liver tissue labeling CD14 with ab183322 at 1/100.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [SP192] (AB183322)

Immunohistochemical analysis of paraffin embedded Human spleen tissue labeling CD14 with ab183322 at 1/100.

• Flow Cyt

Unknown

Flow Cytometry - Anti-CD14 antibody [SP192] (AB183322)

Flow Cytometry analysis of human PBMC (human peripheral blood mononuclear cell) cells labeling CD14 with purified ab183322 at 1 : 200 dilution (0.805 µg/ml) - Red. A Goat anti rabbit IgG (Dylight® 488, ab98462) secondary antibody was used at 1 : 2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) - Black. Unlabeled control - Blue.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [SP192] (AB183322)

Immunohistochemical analysis of paraffin embedded Human lung tissue labeling CD14 with ab183322 at 1/100.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [SP192] (AB183322)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon tissue sections labeling CD14 with ab183322 at 1/100 dilution (1.61 μg/ml). Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 10mins Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. Positive staining on stromal cells in the human colon, performed on a Leica Biosystems BOND™ RX instrument.
The section was incubated with ab183322 for 10 mins at room temperature.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [SP192] (AB183322)

Immunohistochemical analysis of paraffin embedded Human HK lymphoma tissue labeling CD14 with ab183322 at 1/100.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [SP192] (AB183322)

Immunohistochemical analysis of paraffin embedded Human B cell lymphoma tissue labeling CD14 with ab183322 at 1/100.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [SP192] (AB183322)

Immunohistochemical analysis of paraffin embedded Human colon tissue labeling CD14 with ab183322 at 1/100.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [SP192] (AB183322)

Immunohistochemical analysis of paraffin embedded Human bladder tissue labeling CD14 with ab183322 at 1/100.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [SP192] (AB183322)

Immunohistochemical analysis of paraffin embedded Human bone marrow tissue labeling CD14 with ab183322 at 1/100.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [SP192] (AB183322)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human tonsil tissue sections labeling CD14 with ab183322 at 1/100 dilution (1.61 μg/ml). Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 10mins Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. Positive staining on the human tonsil, performed on a Leica Biosystems BOND™ RX instrument.
The section was incubated with ab183322 for 10 mins at room temperature.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [SP192] (AB183322)

Immunohistochemical analysis of paraffin embedded Human endometrium tissue labeling CD14 with ab183322 at 1/100.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [SP192] (AB183322)

Immunohistochemical analysis of paraffin embedded Human colon adenocarcinoma tissue labeling CD14 with ab183322 at 1/100.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD14 antibody [SP192] (AB183322)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human spleen tissue staining CD42d/GP5 with ab322703 at a 1/500 dilution, ab255296 anti-CD177 used at 1/2000 dilution and ab183322 anti-CD14 used at a 1/100 dilution.

Panel A : anti-CD42d/GP5 (green; Opal™520), anti-CD177 (magenta; Opal™690) and anti-CD14 (gray; Opal™570) on human spleen.

Panel B : anti-CD42d/GP5 staining platelets in human spleen.

Panel C : anti-CD177 staining neutrophils in human spleen.

Panel D : anti-CD14 staining blood vessel in human spleen.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab322703, ab255296 and ab183322 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.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [SP192] (AB183322)

Immunohistochemical analysis of formalin-fixed paraffin-embedded human tonsil labelling CD14 with ab230903 at a concentration of 0.5µg/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 32min with ULTRA cell conditioning solution (CC1 pH8.5). ab220903 anti CD14 antibody was incubated at 37°C for 16min. Sections were counterstained is with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

This data was developed using the same antibody clone in a different buffer formulation (ab230903).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [SP192] (AB183322)

Immunohistochemical analysis of paraffin embedded Human thymus tissue labeling CD14 with ab183322 at 1/100.

• WB

Lab

Western blot - Anti-CD14 antibody [SP192] (AB183322)

This blot was produced using 4-20% SDS-PAGE containing 15 μg of human tonsil lysate per lane at 150V for 1hr before being transferred onto a 0.45 μm PVDF membrane at 75V for 1hr. The membrane was then blocked for 1hr using 5% NFDM/TBST, then incubated with ab183322 (1/1000) at room temperature for 1hr. After being washed three times in TBST, the membrane was incubated with Peroxidase conjugated goat anti-rabbit IgG with minimal cross-reactivity with human IgG at 1/2000 dilution for 1hr at room temperature. The membrane was washed three times again. Then the signal was developed using the ECL technique. ab183322 was stored at a range of temperatures (+4°C, +22°C, +37°C) for 1 week before being tested in WB. The image shows the band intensity remains relatively constant across all storage temperatures, demonstrating that antibody activity is not affected.

All lanes:

Western blot - Anti-CD14 antibody [SP192] (ab183322) at 1/1000 dilution

All lanes:

Human tonsyl lysate at 15 µg with NDFM/TBST

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/2000 dilution

false

Exposure time: 20s

• WB

Supplier Data

Western blot - Anti-CD14 antibody [SP192] (AB183322)

All lanes:

Western blot - Anti-CD14 antibody [SP192] (ab183322) at 1/100 dilution

All lanes:

Human tonsil lysate

Predicted band size: 40 kDa

false