Anti-GRP94 antibody [EPR3988]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP94 antibody [EPR3988] (AB108606)

ab108606 staining GRP94 in paraffin embedded Human lung carcinoma tissue sections by Immunohistochemistry (IHC-P- paraformaldehyde-fixed, paraffin embedded sections). Samples are incubated in primary antibody at 1 : 1000 dilution (0.57μg/ml). A ready to use Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Hematoxylin was used as a counterstain. Cytoplasmic staining on human lung carcinoma.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP94 antibody [EPR3988] (AB108606)

ab108606, at 1/100 dilution, staining GRP94 in paraffin-embedded Human gastric adenocarcinoma tissue by Immunohistochemistry.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP94 antibody [EPR3988] (AB108606)

ab108606 staining GRP94 in paraffin embedded Human hepatocellular carcinoma tissue sections by Immunohistochemistry (IHC-P- paraformaldehyde-fixed, paraffin embedded sections). Samples are incubated in primary antibody at 1 : 1000 dilution (0.57μg/ml). A ready to use Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Hematoxylin was used as a counterstain. Cytoplasmic staining on human hepatocellular carcinoma.

• WB

Lab

Western blot - Anti-GRP94 antibody [EPR3988] (AB108606)

Lanes 1-3 : Merged signal (red and green). Green - ab108606 observed at 94 kDa. Red - loading control ab8245 observed at 36 kDa.

ab108606 Anti-GRP94 antibody [EPR3988] was shown to specifically react with GRP94 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266313 (knockout cell lysate ab257254) was used. Wild-type and GRP94 knockout samples were subjected to SDS-PAGE. ab108606 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-GRP94 antibody [EPR3988] (ab108606) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

HSP90B1 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human HSP90B1 (GRP94) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-hsp90b1-grp94-knockout-hek-293t-cell-line-ab266313'>ab266313</a>)

Lane 3:

HeLa cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 92 kDa

Observed band size: 94 kDa

false

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP94 antibody [EPR3988] (AB108606)

ab108606, at 1/100 dilution, staining GRP94 in paraffin-embedded Human breast tissue by Immunohistochemistry.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP94 antibody [EPR3988] (AB108606)

ab108606 staining GRP94 in paraffin embedded Mouse liver carcinoma tissue sections by Immunohistochemistry (IHC-P- paraformaldehyde-fixed, paraffin embedded sections). Samples are incubated in primary antibody at 1 : 1000 dilution (0.57μg/ml). A ready to use Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Hematoxylin was used as a counterstain. Cytoplasmic staining on mouse liver.

• WB

Unknown

Western blot - Anti-GRP94 antibody [EPR3988] (AB108606)

All lanes:

Western blot - Anti-GRP94 antibody [EPR3988] (ab108606) at 1/1000 dilution

Lane 1:

T47D cell lysate at 10 µg

Lane 2:

HeLa cell lysate at 10 µg

Lane 3:

K562 cell lysate at 10 µg

Lane 4:

Jurkat cell lysate at 10 µg

Lane 5:

HepG2 cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 92 kDa

Observed band size: 94 kDa

false

• WB

CiteAb

Western blot - Anti-GRP94 antibody [EPR3988] (AB108606)

GRP94 western blot using anti-GRP94 antibody [EPR3988] ab108606. Publication image and figure legend from Burrello, J., Biemmi, V., et al., 2020, Sci Rep, PubMed 32999414.

ab108606 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab108606 please see the product overview.

EV isolation and purification from plasma sample. (a) Schematic of EV isolating protocol; after serial centrifugations step to remove intact cells, cellular debris and large particles, free-platelet plasma underwent two-step ultracentrifugation to precipitate extracellular vesicles. (b) Representative cumulative distribution plot at nanoparticle tracking analysis for plasma (yellow curve) and EV (blue curve). (c) Western blot analysis for TSG101, as EV specific intraluminal marker, and potential contaminants (GRP94, apolipoprotein A1, and albumin) in representative patients (n = 7), as compared to whole plasma. Level of expression are reported as arbitrary unit (a.u.) in 30 μg of total protein EV lysate. A representative immunoblot is shown referring to STEMI patient at pre-PCI evaluation. Uncropped membranes are shown in Fig. S1a,b.

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