Anti-Glutamine Synthetase 抗体 [EPR13022(B)]

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Glutamine Synthetase antibody [EPR13022(B)] (AB176562)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human retina tissue labeling PAX6, Glutamine Synthetase and CRX with ab109233 at 1/10000 dilution, ab240193 at 1/20000 dilution and ab248897 at 1/1000 dilution followed by a ready to use Opal Polymer HRP Ms + Rb secondary antibody. Nuclear counter stain used was DAPI. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins Panel A : merged staining of anti-CRX (gray; Opal™690), anti-Glutamine Synthetase (green; Opal™520) and anti-PAX6 (red; Opal™570) on human retina. Panel B : anti-PAX6 stained on retinal progenitor cells. Panel C : anti-Glutamine Synthetase stained on Müller glia. Panel D : anti-CRX stained on subset cells of outer nuclear layer and inner nuclear layer. The section was incubated in three rounds of staining : in the order of ab248897, ab240193, and ab109233 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-Glutamine Synthetase antibody [EPR13022(B)] (AB176562)

Immunohistochemical analysis of formalin-fixed, paraffin-embedded, Human glioma tissue labeling Glutamine Synthetase with unpurified ab176562 at a 1/100 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glutamine Synthetase antibody [EPR13022(B)] (AB176562)

Immunohistochemical analysis of formalin-fixed, paraffin-embedded, Human liver tissue labeling Glutamine Synthetase with unpurified ab176562 at a 1/100 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glutamine Synthetase antibody [EPR13022(B)] (AB176562)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human glioma tissue sections labeling Glutamine Synthetase with purified ab176562 at 1 : 500 dilution (0.18 μg/ml). Heat mediated antigen retrieval was performed using citrate Buffer, pH6.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1 : 0 dilution. PBS instead of the primary antibody was used as the negative control.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Glutamine Synthetase antibody [EPR13022(B)] (AB176562)

Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100-permeabilised mouse cerebrum tissue staining glutamine synthetase with ab176562 at 1/250 dilution, followed by alexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution. Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). DAPI was used as a nuclear counterstain.

Positive staining on mouse cerebrum (PMID : 23895693).

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Glutamine Synthetase antibody [EPR13022(B)] (AB176562)

Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100-permeabilised rat cerebrum tissue staining glutamine synthetase with ab176562 at 1/250 dilution, followed by alexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution. Heat mediated antigen retrieval was performed using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). DAPI was used as a nuclear counterstain.

Positive staining on rat cerebrum (PMID : 23895693).

• IHC-P

AbReview62809****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glutamine Synthetase antibody [EPR13022(B)] (AB176562)

ab176562 staining Glutamine Synthetasein Mouse Liver tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were fixed by perfusion with formaldehyde, blocked with PB ab64226 for 10 minutes at room temperature and antigen retrieval was by heat mediation in citrate buffer. The sample was incubated with primary antibody (1/200) for 30 minutes. A HRP-conjugated Goat anti-rabbit polyclonal (1/200) was used as the secondary antibody.

This image is courtesy of Alex Van Engelenburg

• WB

Lab

Western blot - Anti-Glutamine Synthetase antibody [EPR13022(B)] (AB176562)

Blocking and diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Glutamine Synthetase antibody [EPR13022(B)] (ab176562) at 1/2000 dilution

Lane 1:

Human fetal liver lysates at 20 µg

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 15 µg

Lane 3:

Mouse spleen lysates at 20 µg

Lane 4:

Rat spleen lysates at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 42 kDa

Observed band size: 42 kDa

false

• WB

Lab

Western blot - Anti-Glutamine Synthetase antibody [EPR13022(B)] (AB176562)

Lanes 1 - 2 : Merged signal (red and green). Green - ab176562 observed at 42 kDa. Red - loading control, ab130007, observed at 130 kDa.

ab176562 was shown to recognize Glutamine Synthetase in wild-type HAP1 cells as signal was lost at the expected MW in GLUL knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and GLUL knockout samples were subjected to SDS-PAGE. ab176562 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Glutamine Synthetase antibody [EPR13022(B)] (ab176562) at 1 µg

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

GLUL knockout HAP1 whole cell lysate at 20 µg

Predicted band size: 42 kDa

false

• IHC-Fr

AbReview6919e741f4ec5e9fb5f08989****

Immunohistochemistry (Frozen sections) - Anti-Glutamine Synthetase antibody [EPR13022(B)] (AB176562)

Immunohistochemical analysis of PFA-fixed unpermeabilized frozen Nothobranchius furzeri Retina, 6 weeks old (fresh frozen) tissue labeling Glutamine Synthetase with ab176562 at 1/100 dilution for 16 hours at 4°C followed by Goat anti Rabbit Alexa Fluor^® 488 at 1/1000 dilution (pseudocolored to cyan). 1% BSA used as blocking agent for 1 hour.

Antigen retrieval was used prior to antibody labelling. Slides were boiled in NaCit (pH=6.0) for 20 minutes and washed 3x PBS with Triton X (0.1%).

This image is courtesy of an Abreview submitted by Ryan Macdonald

• WB

Supplier Data

Western blot - Anti-Glutamine Synthetase antibody [EPR13022(B)] (AB176562)

All lanes:

Western blot - Anti-Glutamine Synthetase antibody [EPR13022(B)] (ab176562) at 1/1000 dilution

Lane 1:

Human fetal liver lysate at 10 µg

Lane 2:

Jurkat cell lysate at 10 µg

Lane 3:

HeLa cell lysate at 10 µg

Predicted band size: 42 kDa

true