Anti-HYAL2 antibody [EPR28917-156] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HYAL2 antibody [EPR28917-156] - BSA and Azide free (AB323610)

This data was developed using ab323609, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling HYAL2 with ab323609 at 1/100 (4.89 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelial cells of human spleen. The section was incubated with ab323609 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-HYAL2 antibody [EPR28917-156] - BSA and Azide free (AB323610)

This data was developed using ab323609, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human placenta tissue labeling HYAL2 with ab323609 at 1/100 (4.89 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelial cells of human placenta. The section was incubated with ab323609 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-HYAL2 antibody [EPR28917-156] - BSA and Azide free (AB323610)

This data was developed using ab323609, the same antibody clone in a different buffer formulation.

Panel A : anti-CD42d/GP5 (green; Opal™520), anti-CD177 (magenta; Opal™690) and anti-HYAL2 (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-HYAL2 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 ab323609 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-HYAL2 antibody [EPR28917-156] - BSA and Azide free (AB323610)

This data was developed using ab323609, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human cervical carcinoma tissue labeling HYAL2 with ab323609 at 1/100 (4.89 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelial cells of human cervical carcinoma. The section was incubated with ab323609 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-HYAL2 antibody [EPR28917-156] - BSA and Azide free (AB323610)

This data was developed using ab323609, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 100% methanol-fixed, 0.1% Tween-20 permeabilized Saos-2 (human osteosarcoma epithelial cell) cells labeling HYAL2 with ab323609 at 1/50 (9.78 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green).

Confocal image showing partial membranous staining in Saos-2 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue). 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 2ug/ml dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HYAL2 antibody [EPR28917-156] - BSA and Azide free (AB323610)

This data was developed using ab323609, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human lung carcinoma tissue labeling HYAL2 with ab323609 at 1/100 (4.89 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelial cells of human lung carcinoma. The section was incubated with ab323609 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-HYAL2 antibody [EPR28917-156] - BSA and Azide free (AB323610)

This data was developed using ab323609, the same antibody clone in a different buffer formulation.

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

The samples are unboiled.

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

All lanes:

Western blot - Anti-HYAL2 antibody [EPR28917-156] (<a href='/en-us/products/primary-antibodies/hyal2-antibody-epr28917-156-ab323609'>ab323609</a>) at 1/1000 dilution

Lane 1:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

SW480 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

JEG-3 (human placenta epithelial cell) 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: 54 kDa,36 kDa

false

Exposure time: 136s

• WB

Supplier Data

Western blot - Anti-HYAL2 antibody [EPR28917-156] - BSA and Azide free (AB323610)

This data was developed using ab323609, the same antibody clone in a different buffer formulation.

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

The samples are unboiled.

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

All lanes:

Western blot - Anti-HYAL2 antibody [EPR28917-156] (<a href='/en-us/products/primary-antibodies/hyal2-antibody-epr28917-156-ab323609'>ab323609</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HeLa transfected with siRNA specifically targeting HYAL2 whole cell lysate at 20 µg

Lane 3:

Saos-2 (human osteosarcoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 54 kDa,36 kDa

false

Exposure time: 136s

• WB

Supplier Data

Western blot - Anti-HYAL2 antibody [EPR28917-156] - BSA and Azide free (AB323610)

This data was developed using ab323609, the same antibody clone in a different buffer formulation.

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

The samples are unboiled.

All lanes:

Western blot - Anti-HYAL2 antibody [EPR28917-156] (<a href='/en-us/products/primary-antibodies/hyal2-antibody-epr28917-156-ab323609'>ab323609</a>) at 1/1000 dilution

Lane 1:

Human lung cancer tissue lysate at 40 µg

Lane 2:

Human testis tissue lysate at 40 µg

Lane 3:

Human lung tissue lysate at 40 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 1/2000 dilution

Observed band size: 54 kDa

false

Exposure time: 15s