Anti-CD34 antibody [EPR27432-54] - BSA and Azide free

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD34 antibody [EPR27432-54] - BSA and Azide free (AB315803)

This data was developed using ab317582, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human tonsil tissue staining eNOS with ab317582 at a 1 : 500 (1.032 ug/ml) dilution, ab315802 anti-CD34 used at 1 : 2000 (0.26 ug/ml) dilution.

Panel A : merged staining of anti-eNOS (magenta; Opal™520), anti-CD34 (green; Opal™570) on human tonsil.
Panel B : anti-eNOS staining endothelium in human tonsil.
Panel C : anti-CD34 staining endothelium in human tonsil.
Panel D : Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab317582 and ab315802 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD34 antibody [EPR27432-54] - BSA and Azide free (AB315803)

This data was developed using ab315802, the same antibody clone in a different buffer formulation.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil labelling eNOS with ab317582 at 1/500 (B) and CD34 with ab315802 at 1/2000 dilution (C). Opal Polymer HRP Ms + Rb was used as a secondary antibody. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-eNOS (magenta; Opal™520), anti-CD34 (green; Opal™570) on human tonsil.

Panel B : anti-eNOS staining endothelium in human tonsil.

Panel C : anti-CD34 staining endothelium in human tonsil.

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

The section was incubated in three rounds of staining : in the order of ab317582 and ab315802 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-CD34 antibody [EPR27432-54] - BSA and Azide free (AB315803)

This data was developed using ab315802, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling CD34 with ab315802 at 1/2000 (0.262 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelium of human breast cancer (PMID : 25803686). The section was incubated with ab315802 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

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD34 antibody [EPR27432-54] - BSA and Azide free (AB315803)

This data was developed using ab315802, the same antibody clone in a different buffer formulation.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver labelling eNOS with ab317582 at 1/500 (B) and CD34 with ab315802 at 1/2000 dilution (C). Opal Polymer HRP Ms + Rb was used as a secondary antibody. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-eNOS (magenta; Opal™520), anti-CD34 (green; Opal™570) on human liver.

Panel B : anti-eNOS staining endothelium in human liver.

Panel C : anti-CD34 staining endothelium in human liver.

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

The section was incubated in three rounds of staining : in the order of ab317582 and ab315802 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-CD34 antibody [EPR27432-54] - BSA and Azide free (AB315803)

This data was developed using ab315802, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human lung tissue labeling CD34 with ab315802 at 1/2000 (0.262 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelium of human lung (PMID : 16234507). The section was incubated with ab315802 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

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD34 antibody [EPR27432-54] - BSA and Azide free (AB315803)

This data was developed using ab315802, the same antibody clone in a different buffer formulation.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human stomach labelling eNOS with ab317582 at 1/500 (B) and CD34 with ab315802 at 1/2000 dilution (C). Opal Polymer HRP Ms + Rb was used as a secondary antibody. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-eNOS (magenta; Opal™520), anti-CD34 (green; Opal™570) on human stomach.
Panel B : anti-eNOS staining endothelium in human stomach.
Panel C : anti-CD34 staining endothelium in human stomach.
Panel D : Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab317582 and ab315802 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.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD34 antibody [EPR27432-54] - BSA and Azide free (AB315803)

This data was developed using ab315802, the same antibody clone in a different buffer formulation.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human endometrium labelling eNOS with ab317582 at 1/500 (B) and CD34 with ab315802 at 1/2000 dilution (C). Opal Polymer HRP Ms + Rb was used as a secondary antibody. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-eNOS (magenta; Opal™520), anti-CD34 (green; Opal™570) on human endometrium.
Panel B : anti-eNOS staining endothelium in human endometrium.
Panel C : anti-CD34 staining endothelium in human endometrium.
Panel D : Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab317582 and ab315802 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.

The section was incubated in three rounds of staining : in the order of ab317582 and ab315802 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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CD34 antibody [EPR27432-54] - BSA and Azide free (AB315803)

This data was developed using ab315802, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized KG-1a (human bone marrow myeloblast) cells labelling CD34 with ab315802 at 1/500 (1.048 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 membranous staining in KG-1a cell line.
Negative control : Daudi (PMID : 10942240)
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-CD34 antibody [EPR27432-54] - BSA and Azide free (AB315803)

This data was developed using ab315802, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of Daudi (human Burkitt's lymphoma lymphoblast, Left) / KG-1a (human bone marrow myeloblast, Right) cells labelling CD34 with ab315802 at 1/500 dilution (0.1 ug)/Red compared with a Rabbit monoclonal IgG (ab172730) (Black) 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.

Negative control : Daudi (PMID : 21286385, 10942240). Gated on viable cells.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD34 antibody [EPR27432-54] - BSA and Azide free (AB315803)

This data was developed using ab317582, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human liver tissue staining eNOS with ab317582 at a 1 : 500 (1.032 ug/ml) dilution, ab315802 anti-CD34 used at 1 : 2000 (0.26 ug/ml) dilution.

Panel A : merged staining of anti-eNOS (magenta; Opal™520), anti-CD34 (green; Opal™570) on human liver.
Panel B : anti-eNOS staining endothelium in human liver.
Panel C : anti-CD34 staining endothelium in human liver.
Panel D : Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab317582 and ab315802 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 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-CD34 antibody [EPR27432-54] - BSA and Azide free (AB315803)

This data was developed using ab315802, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human placenta tissue labeling CD34 with ab315802 at 1/2000 (0.262 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelium of human placenta. The section was incubated with ab315802 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

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD34 antibody [EPR27432-54] - BSA and Azide free (AB315803)

This data was developed using ab315802, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human placenta tissue staining CD68 with ab213363 at a 1/500 dilution, ab315802 anti-CD34 used at 1/2000 dilution and ab255296 anti-CD177 used at a 1/2000 dilution.

Panel A : merged staining of anti-CD68 (green; Opal™520), anti-CD34 (magenta; Opal™690) and anti-CD177 (gray; Opal™570) on human placenta.

Panel B : anti-CD68 staining macrophages in human placenta.

Panel C : ant-CD34 staining endothelium in human placenta.

Panel D : ant-CD177 staining neutrophils in human placenta.

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

The section was incubated in three rounds of staining : in the order of ab213363, ab315802 and ab255296 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• WB

Supplier Data

Western blot - Anti-CD34 antibody [EPR27432-54] - BSA and Azide free (AB315803)

This data was developed using ab315802, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Negative control : Daudi (PMID : 21286385, 10942240), HuT-78.

The molecular weight observed is consistent with what has been described in the literature (PMID : 37181754, 29296932).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-CD34 antibody [EPR27432-54] (<a href='/en-us/products/primary-antibodies/cd34-antibody-epr27432-54-ab315802'>ab315802</a>) at 1/1000 dilution

Lane 1:

KG-1a (human bone marrow myeloblast) whole cell lysate at 20 µg with NFDM/TBST

Lane 2:

Daudi (human Burkitts lymphoma lymphoblast) whole cell lysate at 20 µg with NFDM/TBST

Lane 3:

HuT-78 (human Sezary syndrome cutaneous T lymphocyte) whole cell lysate at 20 µg with NFDM/TBST

Lane 4:

TF-1 (human erythroleukemia erythroblast) whole cell lysate at 20 µg with NFDM/TBST

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 1/100000 dilution

Observed band size: 120 kDa,36 kDa

true

Exposure time: 81s

• WB

Supplier Data

Western blot - Anti-CD34 antibody [EPR27432-54] - BSA and Azide free (AB315803)

This data was developed using ab315802, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The molecular weight observed is consistent with what has been described in the literature (PMID : 37181754, 29296932).

CD34 is a glycoprotein of approximately 120 kDa and detected as a 80-90 kDa band after treated with Protein Deglycosylation Peptide : N-glycosidase F (PNGase F).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-CD34 antibody [EPR27432-54] (<a href='/en-us/products/primary-antibodies/cd34-antibody-epr27432-54-ab315802'>ab315802</a>) at 1/1000 dilution

Lane 1:

Untreated human lung tissue lysate at 20 µg with NFDM/TBST

Lane 2:

Human lung tissue lysate treated with Protein Deglycosylation Peptide: N-glycosidase F (PNGase F) at 20 µg with NFDM/TBST

Lane 3:

Untreated human placenta tissue lysate at 20 µg with NFDM/TBST

Lane 4:

Human placenta tissue lysate treated with Protein Deglycosylation Peptide: N-glycosidase F (PNGase F) at 20 µg with NFDM/TBST

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 1/100000 dilution

Observed band size: 120 kDa,36 kDa

false

Exposure time: 180s