Anti-SH3GL3 antibody [EPR29565-553]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SH3GL3 antibody [EPR29565-553] (AB323569)

Immunohistochemical analysis of paraffin-embedded Human astrocytoma tissue labeling SH3GL3 with ab323569 at 1/100 (4.9 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in human astrocytoma.
The section was incubated with ab323569 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-SH3GL3 antibody [EPR29565-553] (AB323569)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling SH3GL3 with ab323569 at 1/100 (4.9 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining in human liver (PMID : 9809064).
The section was incubated with ab323569 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-SH3GL3 antibody [EPR29565-553] (AB323569)

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling SH3GL3 with ab323569 at 1/100 (4.9 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in mouse testis.
The section was incubated with ab323569 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-SH3GL3 antibody [EPR29565-553] (AB323569)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling SH3GL3 with ab323569 at 1/100 (4.9 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining in mouse liver (PMID : 9809064).
The section was incubated with ab323569 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-SH3GL3 antibody [EPR29565-553] (AB323569)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling SH3GL3 with ab323569 at 1/100 (4.9 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in mouse cerebrum.
The section was incubated with ab323569 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

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Immunocytochemistry/ Immunofluorescence - Anti-SH3GL3 antibody [EPR29565-553] (AB323569)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Neuro-2a (mouse neuroblastoma neuroblast) cells labelling SH3GL3 with ab323569 at 1/100 (4.9 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 cytoplasmic and membranous staining in Neuro-2a cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Negative control : A20.

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.

• WB

Supplier Data

Western blot - Anti-SH3GL3 antibody [EPR29565-553] (AB323569)

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

Negative control : liver (PMID : 9809064)

Low expression : HeLa, SU-DHL-1.

Lanes 3-8 are incubated with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 and lanes 1-2 are incubated with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000.

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

All lanes:

Western blot - Anti-SH3GL3 antibody [EPR29565-553] (ab323569) at 1/1000 dilution

Lane 1:

Human striatum tissue lysate at 20 µg

Lane 2:

Human liver tissue lysate at 20 µg

Lane 3:

SK-N-MC (human brain epithelial cell) whole cell lysate at 20 µg

Lane 4:

HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 5:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Lane 6:

A-673 (human muscle ewings sarcoma cell line) whole cell lysate at 20 µg

Lane 7:

SU-DHL-1 (human histiocytic lymphoblast-like cell) whole cell lysate at 20 µg

Lane 8:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

Lanes 1 - 2:

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

Lanes 3 - 8:

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: 39 kDa,32 kDa,36 kDa

false

Exposure time: 59s

• WB

Supplier Data

Western blot - Anti-SH3GL3 antibody [EPR29565-553] (AB323569)

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

Negative control : heart, liver (PMID : 9809064).

Low expression : A20, B16-F10.

The identity of the bands higher than 50 kDa are unknown.

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

All lanes:

Western blot - Anti-SH3GL3 antibody [EPR29565-553] (ab323569) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse testis tissue lysate at 20 µg

Lane 3:

Mouse heart tissue lysate at 20 µg

Lane 4:

Mouse liver tissue lysate at 20 µg

Lane 5:

Mouse striatum tissue lysate at 20 µg

Lane 6:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 7:

A20 (mouse reticulum sarcoma b lymphocyte) whole cell lysate at 20 µg

Lane 8:

B16-F10 (mouse skin melanoma 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

false

Exposure time: 48s

• WB

Supplier Data

Western blot - Anti-SH3GL3 antibody [EPR29565-553] (AB323569)

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

This antibody does not cross-react with recombinant human SH3GL2 or SH3GL1 by western blot.

In Western blot, Anti-SH3GL1 + SH3GL2 + SH3GL3 antibody [EPR10215(B)] - (ab150431) - total protein Control staining at 1/1000 dilution.

All lanes:

Western blot - Anti-SH3GL3 antibody [EPR29565-553] (ab323569) at 1/1000 dilution

Lane 1:

Human SH3GL3 recombinant protein (no-tag) at 10 ng

Lane 2:

Human SH3GL2 recombinant protein (no-tag) at 10 ng

Lane 3:

Human SH3GL1 recombinant protein (no-tag) at 10 ng

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: 39 kDa,32 kDa,40 kDa,42 kDa

false

Exposure time: 8s