Anti-CD34 antibody [RM2052]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD34 antibody [RM2052] (AB317588)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling CD34 with ab317588 at 1/2000 (0.247 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelium of human kidney (PMID : 16234507). The section was incubated with ab317588 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD34 antibody [RM2052] (AB317588)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human kidney tissue staining CD34 with ab317588 at a 1/2000 dilution, ab199975 anti-Aquaporin 2 used at 1/4000 dilution and ab212197 anti-Renin used at a 1/4000 dilution.

Panel A : merged staining of anti-CD34 (green; Opal™520), anti-Aquaporin 2 (magenta; Opal™690) and anti-Renin (gray; Opal™570) on human kidney.

Panel B : anti-CD34 staining endothelium in human kidney.

Panel C : anti-Aquaporin 2 staining collecting tubules in human kidney.

Panel D : anti-Renin staining juxtaglomerular cells in human kidney.

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

The section was incubated in three rounds of staining : in the order of ab317588, ab199975 and ab212197 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 solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD34 antibody [RM2052] (AB317588)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human kidney tissue staining CD34 with ab317588 at a 1/2000 dilution, ab321787 anti-SGLT1 used at 1/2000 dilution and ab212197 anti-Renin used at a 1/4000 dilution.

Panel A : merged staining of anti-CD34 (green; Opal™520), anti-SGLT1 (magenta; Opal™690) and anti-Renin (gray; Opal™570) on human kidney.

Panel B : anti-CD34 staining endothelium in human kidney.

Panel C : anti-SGLT1 staining tubules in human kidney.

Panel D : anti-Renin staining juxtaglomerular cells in human kidney.

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

The section was incubated in three rounds of staining : in the order of ab317588, ab321787 and ab212197 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 solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD34 antibody [RM2052] (AB317588)

Immunohistochemical analysis of paraffin-embedded Human colon cancer tissue labeling CD34 with ab317588 at 1/2000 (0.247 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelium of human colon cancer. The section was incubated with ab317588 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD34 antibody [RM2052] (AB317588)

Immunohistochemical analysis of paraffin-embedded Human placenta tissue labeling CD34 with ab317588 at 1/2000 (0.247 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelium of human placenta (PMID : 35263417). The section was incubated with ab317588 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD34 antibody [RM2052] (AB317588)

Flow cytometric analysis of Daudi(human Burkitt's lymphoma lymphoblast, Left) / KG-1a(human bone marrow myeloblast, Right) cells labelling CD34 with ab317588 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.

Gated on viable cell.
Low expression : Daudi.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CD34 antibody [RM2052] (AB317588)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized KG-1a (human bone marrow myeloblast) cells labelling CD34 with ab317588 at 1/50 (9.88 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 KG-1a cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Low expression : Daudi (PMID : 21286385).
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 (Magenta). 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD34 antibody [RM2052] (AB317588)

Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling CD34 with ab317588 at 1/5000 (0.099 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelium of mouse lung. The section was incubated with ab317588 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD34 antibody [RM2052] (AB317588)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling CD34 with ab317588 at 1/5000 (0.099 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelium of mouse kidney. The section was incubated with ab317588 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD34 antibody [RM2052] (AB317588)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded rat kidney tissue staining Renin with ab326049 at a 1 : 5000 (0.103 µg/ml) dilution ab317588 anti-CD34 used at 1 : 5000 (0.099 µg/ml) dilution and ab199975 anti-Aquaporin 2 used at a 1 : 4000 (0.152 µg/ml) dilution.

Panel A : merged staining of anti-Renin (green; Opal™520) anti-CD34 (magenta; Opal™690) and anti-Aquaporin 2 (gray; Opal™570) on rat kidney.
Panel B : anti-Renin staining juxtaglomerular cells in rat kidney.
Panel C : anti-CD34 staining endothelium in rat kidney.
Panel D : anti-Aquaporin 2 staining collecting tubules in rat kidney.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab326049 ab317588 and ab199975 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

Lab

Multiplex immunohistochemistry - Anti-CD34 antibody [RM2052] (AB317588)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse kidney tissue staining Renin with ab326049 at a 1 : 5000 (0.103 µg/ml) dilution ab317588 anti-CD34 used at 1 : 5000 (0.099 µg/ml) dilution and ab199975 anti-Aquaporin 2 used at a 1 : 4000 (0.152 µg/ml) dilution.

Panel A : merged staining of anti-Renin (green; Opal™520) anti-CD34 (magenta; Opal™690) and anti-Aquaporin 2 (gray; Opal™570) on mouse kidney.
Panel B : anti-Renin staining juxtaglomerular cells in mouse kidney.
Panel C : anti-CD34 staining endothelium in mouse kidney.
Panel D : anti-Aquaporin 2 staining collecting tubules in mouse kidney.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab326049 ab317588 and ab199975 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.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD34 antibody [RM2052] (AB317588)

Flow cytometric analysis of Neuro-2a(mouse neuroblastoma neuroblast, Left) / NIH/3T3(mouse embryonic fibroblast, Right) cells labelling CD34 with ab317588 at 1/50 dilution (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.

Gated on viable cell.
Low expression : Neuro-2a.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD34 antibody [RM2052] (AB317588)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling CD34 with ab317588 at 1/5000 (0.099 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelium of rat kidney. The section was incubated with ab317588 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD34 antibody [RM2052] (AB317588)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse kidney tissue staining CD34 with ab317588 at a 1/5000 dilution, ab199975 anti-Aquaporin 2 used at 1/4000 dilution and ab212197 anti-Renin used at a 1/4000 dilution.

Panel A : merged staining of anti-CD34 (green; Opal™520), anti-Aquaporin 2 (magenta; Opal™690) and anti-Renin (gray; Opal™570) on human kidney.

Panel B : anti-CD34 staining endothelium in mouse kidney.

Panel C : anti-Aquaporin 2 staining collecting tubules in mouse kidney.

Panel D : anti-Renin staining juxtaglomerular cells in mouse kidney.

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

The section was incubated in three rounds of staining : in the order of ab317588, ab199975 and ab212197 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 solution2) for 20 mins.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD34 antibody [RM2052] (AB317588)

Flow cytometric analysis of Mouse bone marrow cells labelling CD34 with ab317588 at 1/500 dilution (0.1ug)/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.

Mouse bone marrow are co-stained with CD117 conjugated APC-eFluor 780.
Gated on viable cell.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD34 antibody [RM2052] (AB317588)

Immunohistochemical analysis of paraffin-embedded Rat lung tissue labeling CD34 with ab317588 at 1/5000 (0.099 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelium of rat lung. The section was incubated with ab317588 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• WB

Supplier Data

Western blot - Anti-CD34 antibody [RM2052] (AB317588)

Low expression : Daudi (PMID : 21286385), HuT-78.

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

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

All lanes:

Western blot - Anti-CD34 antibody [RM2052] (ab317588) at 1/1000 dilution

Lane 1:

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

Lane 2:

Daudi (human Burkitts lymphoma lymphoblast) whole cell lysate at 20 µg

Lane 3:

HuT-78 (human Sezary syndrome cutaneous T lymphocyte) 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 1/100000 dilution

Observed band size: 80-120 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-CD34 antibody [RM2052] (AB317588)

Low expression : liver (PMID : 1709048).

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

The identity of the lower MW band at approximately 40 kDa (in lane 3) is unknown.

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

Exposure time : Lane 2 : 10 seconds, lanes 1 and 3 : 180 seconds

All lanes:

Western blot - Anti-CD34 antibody [RM2052] (ab317588) at 1/1000 dilution

Lane 1:

Mouse testis tissue lysate at 20 µg

Lane 2:

Mouse lung tissue lysate at 20 µg

Lane 3:

Mouse liver 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 1/100000 dilution

Observed band size: 80-120 kDa,36 kDa

true

• WB

Supplier Data

Western blot - Anti-CD34 antibody [RM2052] (AB317588)

Low expression : Neuro-2a.

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

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

Exposure time : Lane 1 : 15 seconds, lanes 2-3 : 180 seconds

All lanes:

Western blot - Anti-CD34 antibody [RM2052] (ab317588) at 1/1000 dilution

Lane 1:

bEnd.3 (mouse brain endothelial cell) whole cell lysate at 20 µg

Lane 2:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 3:

Neuro-2a (mouse neuroblastoma neuroblast) 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 1/100000 dilution

Observed band size: 80-120 kDa,36 kDa

false