Anti-CD34 antibody [EPR27431-71] - BSA and Azide free

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD34 antibody [EPR27431-71] - BSA and Azide free (AB316278)

This data was developed using ab316277, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse breast tissue staining CD34 with ab316277 at a 1 : 10000 (0.05 ug/ml) dilution, ab324191 E Cadherin used at 1 : 500 (0.228 ug/ml) dilution and ab316115 Perilipin-1 used at 1 : 2000 (0.251 μg/ml) dilution.

Panel A : merged staining of anti-CD34 (green; Opal™570), anti-E Cadherin (magenta; Opal™690) and anti-Perilipin-1 (gray; Opal™570) on mouse breast.

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

Panel C : ant-E Cadherin staining epithelium in mouse breast.

Panel D : ant-Perilipin-1 staining adipocytes in mouse breast.

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

The section was incubated in three rounds of staining : in the order of ab316277, ab324191 and ab316115 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 Solution2) for 20 mins.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD34 antibody [EPR27431-71] - BSA and Azide free (AB316278)

This data was developed using ab316277, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of Mouse bone marrow cells labelling CD34 with ab316277 at 1/500 dilution (0.1 ug)/Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells. Cells are co-stained with CD117 conjugated to APC-eFluor 780. (PMID : 23295606)

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD34 antibody [EPR27431-71] - BSA and Azide free (AB316278)

This data was developed using ab316277, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of Neuro-2a (mouse neuroblastoma neuroblast, Left) / NIH/3T3 (mouse embryonic fibroblast, Right) cells labelling CD34 with ab316277 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 : Neuro-2a.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD34 antibody [EPR27431-71] - BSA and Azide free (AB316278)

This data was developed using ab322349, the same antibody clone in a different buffer formulation.

Immunohistochemistry analysis of Formalin/PFA-fixed paraffin-embedded sections mouse stomach tissue labelling Sodium/Hydrogen Exchanger 1/NHE-1 with ab322349 at 1/4000 dilution (B), CD34 with ab316277 at 1/10000 dilution (C) and Cathepsin E with ab322657 at 1/5000 dilution (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-Sodium/Hydrogen Exchanger 1/NHE-1 (magenta; Opal™690), anti-CD34 (green; Opal™520) and anti-Cathepsin E (gray; Opal™570) on mouse stomach.

Panel B : anti-Sodium/Hydrogen Exchanger 1/NHE-1 showed membranous staining in epithelium of mouse stomach.

Panel C : anti-CD34 staining endothelium in mouse stomach.

Panel D : anti-Cathepsin E showed cytoplasmic staining in mouse stomach.

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

The section was incubated in three rounds of staining : in the order of ab322349, ab316277 and ab322657 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 [EPR27431-71] - BSA and Azide free (AB316278)

This data was developed using ab316277, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling CD34 with ab316277 at 1/8000 (0.064 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : weak staining on endothelial cells of rat liver. The section was incubated with ab316277 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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-CD34 antibody [EPR27431-71] - BSA and Azide free (AB316278)

This data was developed using ab316277, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling CD34 with ab316277 at 1/8000 (0.064 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelial cells of mouse kidney. The section was incubated with ab316277 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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-CD34 antibody [EPR27431-71] - BSA and Azide free (AB316278)

This data was developed using ab316277, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling CD34 with ab316277 at 1/8000 (0.064 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelial cells of rat kidney. The section was incubated with ab316277 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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-CD34 antibody [EPR27431-71] - BSA and Azide free (AB316278)

This data was developed using ab316277, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling CD34 with ab316277 at 1/8000 (0.064 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : weak staining on endothelial cells of mouse liver (PMID : 1709048). The section was incubated with ab316277 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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-CD34 antibody [EPR27431-71] - BSA and Azide free (AB316278)

This data was developed using ab316277, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling CD34 with ab316277 at 1/8000 (0.064 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on basement membrane of rat testis. The section was incubated with ab316277 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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-CD34 antibody [EPR27431-71] - BSA and Azide free (AB316278)

This data was developed using ab316277, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling CD34 with ab316277 at 1/8000 (0.064 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on basement membrane of mouse testis (PMID : 1709048). The section was incubated with ab316277 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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-CD34 antibody [EPR27431-71] - BSA and Azide free (AB316278)

This data was developed using ab316277, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse squamouse cell carcinoma tissue labeling CD34 with ab316277 at 1/8000 (0.064 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelial cells of mouse squamouse cell carcinoma. The section was incubated with ab316277 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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-CD34 antibody [EPR27431-71] - BSA and Azide free (AB316278)

This data was developed using ab316277, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling CD34 with ab316277 at 1/8000 (0.064 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelial cells of mouse spleen (PMID : 1709048). The section was incubated with ab316277 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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-CD34 antibody [EPR27431-71] - BSA and Azide free (AB316278)

This data was developed using ab316277, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling CD34 with ab316277 at 1/8000 (0.064 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelial cells of rat cerebrum. The section was incubated with ab316277 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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-CD34 antibody [EPR27431-71] - BSA and Azide free (AB316278)

This data was developed using ab316277, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling CD34 with ab316277 at 1/100 (5.09 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 NIH/3T3 cell line. Negative control : Neuro-2a. 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.

• WB

Supplier Data

Western blot - Anti-CD34 antibody [EPR27431-71] - BSA and Azide free (AB316278)

This data was developed using ab316277, the same antibody clone in a different buffer formulation.

Low expression tissue : liver (PMID : 1709048).

The identity of the bands between 25 kDa and 37 kDa in lane 4 are unknown.

The lanes 2-4 were developed using a high sensitivity ECL substrate.

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

Exposure time : Lane 1 : 59 seconds, lanes 2-4 : 180 seconds

All lanes:

Western blot - Anti-CD34 antibody [EPR27431-71] (<a href='/en-us/products/primary-antibodies/cd34-antibody-epr27431-71-ab316277'>ab316277</a>) at 1/1000 dilution

Lane 1:

Mouse lung tissue lysate at 20 µg

Lane 2:

Mouse placenta tissue lysate at 20 µg

Lane 3:

Mouse testis tissue lysate at 20 µg

Lane 4:

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 kDa,36 kDa

true

• WB

Supplier Data

Western blot - Anti-CD34 antibody [EPR27431-71] - BSA and Azide free (AB316278)

This data was developed using ab316277, the same antibody clone in a different buffer formulation.

Negative control : Neuro-2a.

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

All lanes:

Western blot - Anti-CD34 antibody [EPR27431-71] (<a href='/en-us/products/primary-antibodies/cd34-antibody-epr27431-71-ab316277'>ab316277</a>) at 1/1000 dilution

Lane 1:

bEnd.3 (mouse brain endothelioma) 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 kDa,15 kDa

false

Exposure time: 59s

• WB

Supplier Data

Western blot - Anti-CD34 antibody [EPR27431-71] - BSA and Azide free (AB316278)

This data was developed using ab316277, the same antibody clone in a different buffer formulation.

This antibody doesn't detect with de-glycosylated form of CD34.

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

All lanes:

Western blot - Anti-CD34 antibody [EPR27431-71] (<a href='/en-us/products/primary-antibodies/cd34-antibody-epr27431-71-ab316277'>ab316277</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

NIH/3T3 treated with Peptide:N-glycosidase F (PNGase F) 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 kDa,36 kDa

true

Exposure time: 180s