Anti-Sialoadhesin/CD169 antibody [EPR27102-11] - BSA and Azide free

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-Sialoadhesin/CD169 antibody [EPR27102-11] - BSA and Azide free (AB312841)

This data was developed using ab312840, the same antibody clone in a different buffer formulation. Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse spleen tissue labeling Sialoadhesin/CD169, CD3 epsilon and CD20 with ab312840 at 1/100 dilution, ab16669 at 1 : 150 dilution and ab236434 at 1 : 5000 dilution. Panel A : merged staining of anti-Sialoadhesin/CD169 (red; Opal™690), anti-CD3 epsilon (green; Opal™520) and anti-CD20 (gray; Opal™570) on mouse spleen. Panel B : anti-Sialoadhesin/CD169 stained on macrophages. Panel C : anti-CD3 epsilon stained on T cells. Panel D : anti-CD20 stained on B cells. The section was incubated in three rounds of staining : in the order of ab312840, ab16669, and ab236434 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sialoadhesin/CD169 antibody [EPR27102-11] - BSA and Azide free (AB312841)

This data was developed using ab312840, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling Sialoadhesin/CD169 with ab312840 at 1/100 (5.16 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on rat kidney. The section was incubated with ab312840 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). 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-Sialoadhesin/CD169 antibody [EPR27102-11] - BSA and Azide free (AB312841)

This data was developed using ab312840, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Sialoadhesin/CD169 with ab312840 at 1/100 (5.16 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on mouse kidney. The section was incubated with ab312840 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). 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-Sialoadhesin/CD169 antibody [EPR27102-11] - BSA and Azide free (AB312841)

This data was developed using ab312840, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscl tissue labeling Sialoadhesin/CD169 with ab312840 at 1/100 (5.16 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on mouse skeletal muscle. The section was incubated with ab312840 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). 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-Sialoadhesin/CD169 antibody [EPR27102-11] - BSA and Azide free (AB312841)

This data was developed using ab312840, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human tissue labeling Sialoadhesin/CD169 with ab312840 at 1/2000 (0.258 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on (A) HEK-293T transfected with a mouse SIGLEC1 expression vector containing a his tag. No staining on (B) HEK-293T transfected with empty vector containing a his tag. The section was incubated with ab312840 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). 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-Sialoadhesin/CD169 antibody [EPR27102-11] - BSA and Azide free (AB312841)

This data was developed using ab312840, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling Sialoadhesin/CD169 with ab312840 at 1/100 (5.16 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse spleen interfollicular zone (PMID : 15107454). The section was incubated with ab312840 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). 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-Sialoadhesin/CD169 antibody [EPR27102-11] - BSA and Azide free (AB312841)

This data was developed using ab312840, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling Sialoadhesin/CD169 with ab312840 at 1/100 (5.16 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat spleen interfollicular zone. The section was incubated with ab312840 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Sialoadhesin/CD169 antibody [EPR27102-11] - BSA and Azide free (AB312841)

This data was developed using ab312840, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) cells labelling Sialoadhesin/CD169 with ab312840 at 1/250 (2.064 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing positive staining in RAW 264.7 cells treated with lipopolysaccharide (100ng/ml) for 16 hours.Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-Sialoadhesin/CD169 antibody [EPR27102-11] - BSA and Azide free (AB312841)

This data was developed using ab312840, the same antibody clone in a different buffer formulation. Flow cytometric analysis of Mouse bone marrow cells labelling Sialoadhesin/CD169 with ab312840 at 1/50 dilution (1ug)/ Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody. Cells co-stained with CD3 conjugated to A647. Gated on viable cells.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-Sialoadhesin/CD169 antibody [EPR27102-11] - BSA and Azide free (AB312841)

This data was developed using ab312840, the same antibody clone in a different buffer formulation. Flow cytometric analysis of Mouse bone marrow cells labelling Sialoadhesin/CD169 with ab312840 at 1/50 dilution (1ug)/ Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody. Cells co-stained with F4/80 conjugated to PE. Gated on viable cells.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-Sialoadhesin/CD169 antibody [EPR27102-11] - BSA and Azide free (AB312841)

This data was developed using ab312840, the same antibody clone in a different buffer formulation. Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Rat spleen tissue labeling Sialoadhesin/CD169, CD3 epsilon and CD20 with ab312840 at 1/100 dilution, ab16669 at 1 : 150 dilution and ab236434 at 1 : 5000 dilution. Panel A : merged staining of anti-Sialoadhesin/CD169 (red; Opal™690), anti-CD3 epsilon (green; Opal™520) and anti-CD20 (gray; Opal™570) on rat spleen. Panel B : anti-Sialoadhesin/CD169 stained on macrophages. Panel C : anti-CD3 epsilon stained on T cells. Panel D : anti-CD20 stained on B cells. The section was incubated in three rounds of staining : in the order of ab312840, ab16669, and ab236434 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sialoadhesin/CD169 antibody [EPR27102-11] - BSA and Azide free (AB312841)

This data was developed using ab312840, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Rat skeletal muscle tissue labeling Sialoadhesin/CD169 with ab312840 at 1/100 (5.16 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on rat skeletal muscle. The section was incubated with ab312840 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• WB

Supplier Data

Western blot - Anti-Sialoadhesin/CD169 antibody [EPR27102-11] - BSA and Azide free (AB312841)

This data was developed using ab312840, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST The expression of Sialoadhesin/CD169 is upregulated in response to LPS treatment (PMID : 27129263). In Western blot, anti- Vinculin antibody (ab129002) loading control staining at 1/10000 dilution. Exposure time : 180 seconds

All lanes:

Western blot - Anti-Sialoadhesin/CD169 antibody [EPR27102-11] (<a href='/en-us/products/primary-antibodies/sialoadhesin-cd169-antibody-epr27102-11-ab312840'>ab312840</a>) at 1/1000 dilution

Lane 1:

Untreated RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 2:

RAW 264.7 treated with /ml LPS for 16h whole cell 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

Observed band size: 250 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Sialoadhesin/CD169 antibody [EPR27102-11] - BSA and Azide free (AB312841)

This data was developed using ab312840, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Low expression : kidney (PMID 33408860). This blot of Lane 6 was developed using a high sensitivity ECL substrate. In Western blot, anti- Vinculin antibody (ab129002) loading control staining at 1/10000 dilution. Exposure time : Lane 1-5 : 180 seconds; Lane 6 : 114 seconds.

All lanes:

Western blot - Anti-Sialoadhesin/CD169 antibody [EPR27102-11] (<a href='/en-us/products/primary-antibodies/sialoadhesin-cd169-antibody-epr27102-11-ab312840'>ab312840</a>) at 1/1000 dilution

Lane 1:

Mouse spleen tissue lysate at 20 µg

Lane 2:

Mouse lymph node tissue lysate at 20 µg

Lane 3:

Mouse kidney tissue lysate at 20 µg

Lane 4:

Rat lymph node tissue lysate at 20 µg

Lane 5:

Rat kidney tissue lysate at 20 µg

Lane 6:

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

Observed band size: 250 kDa

false