MA3-016

antibody from Invitrogen Antibodies

Targeting: HSP90B1 GP96, GRP94, TRA1

Provider product page for MA3-016

Western blot

Immunocytochemistry

Immunoprecipitation

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [27]
Comments [0]
Validations
Immunocytochemistry [8]
Immunohistochemistry [4]
Other assay [4]

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Product number: MA3-016 - Provider product page
Provider: Invitrogen Antibodies
Product name: GRP94 Monoclonal Antibody (9G10)
Antibody type: Monoclonal
Antigen: Other
Description: MA3-016 detects glucose regulated protein 94 kDa (GRP94) from human, rat, mouse, chick, hamster and bovine tissues. MA3-016 is also expected to react with GRP94 from Xenopus, human, canine, rabbit, non-human primate, canine, Drosophila, and ovine tissues. MA3-016 has been successfully used in Western blot, Immunohistochemistry (paraffin), immunocytochemistry, immunofluorescence and immunoprecipitation procedures. By Western blot, this antibody detects an ~108 kDa protein representing GRP94 in chick oviduct cytosol. The MA3-016 immunogen is chick oviduct GRP94.
Reactivity: Human, Mouse, Rat, Bovine, Chicken/Avian, Hamster
Host: Rat
Isotype: IgG
Antibody clone number: 9G10
Vial size: 100 µL
Concentration: Conc. Not Determined
Storage: -20° C, Avoid Freeze/Thaw Cycles

HSP90B1 protein structure - MA3-016 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of GRP94 using a monoclonal antibody (Product # MA3-016).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Glucose-Regulated Protein 94 using Glucose-Regulated Protein 94 Monoclonal Antibody (9G10) (Product # MA3-016) shows staining in C6 Cells. Glucose-Regulated Protein 94 (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with an antibody recognizing Glucose-Regulated Protein 94 (Product # MA3-016) at a dilution of 1:100 over night at 4ºC, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Glucose-Regulated Protein 94 using Glucose-Regulated Protein 94 Monoclonal Antibody (9G10) (Product # MA3-016) shows staining in Hela Cells. Glucose-Regulated Protein 94 (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with an antibody recognizing Glucose-Regulated Protein 94 (Product # MA3-016) at a dilution of 1:100 over night at 4ºC, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of GRP94 using a monoclonal antibody (Product # MA3-016).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of GRP94 using a monoclonal antibody (Product # MA3-016).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry analysis of Glucose-Regulated Protein 94 (Grp94) in U2OS cells. Cells fixed with 4% paraformaldehyde were permeabilized with 0.1% Triton X-100 (Product # 28314) in PBS for 15 minutes at room temperature and blocked with 2% BSA in PBST (from Product # 37525) for 30 minutes at room temperature. Cells were treated with Peroxidase Suppressor (Product # 35000), probed with a Grp94 monoclonal antibody (Product # MA3-016) at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBST, and incubated with an HRP-conjugated goat anti-rat IgG (H+L) secondary antibody (Product # 31471) at a dilution of 1:1000 for 30 minutes at room temperature. Chromogenic detection was performed using Metal Enhanced DAB Substrate Kit (Product # 34065). Images were taken on a Zeiss Axio Observer microscope at 20X magnification (x1.6 Optovar ~ 32X).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Glucose-Regulated Protein 94 using Glucose-Regulated Protein 94 Monoclonal Antibody (9G10) (Product # MA3-016) shows staining in NIH-3T3 Cells. Glucose-Regulated Protein 94 (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with an antibody recognizing Glucose-Regulated Protein 94 (Product # MA3-016) at a dilution of 1:20 over night at 4ºC, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of GRP94 using a monoclonal antibody (Product # MA3-016).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on normal biopsies of deparaffinized Mouse liver tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a mouse monoclonal antibody recognizing Glucose Regulated Protein 94 (Product # MA3-016) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on normal biopsies of deparaffinized Mouse breast tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a mouse monoclonal antibody recognizing Glucose Regulated Protein 94 (Product # MA3-016) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on normal biopsies of deparaffinized Mouse lymph node. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a mouse monoclonal antibody recognizing Glucose Regulated Protein 94 (Product # MA3-016) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on poorly differentiated adenocarcinoma from paraffin-embedded human colon tissue sections. To expose target proteins, heat-induced epitope retrieval was performed using 10mM sodium citrate (pH 6.0) buffer for 20 minutes at 95ºC. Following antigen retrieval, tissues were blocked in 3% BSA (Product # 37525) in PBST for 30 minutes at room temperature and then probed with a Grp94 monoclonal antibody (Product # MA3-016) at a dilution of 1:100 for 1 hour in a humidified chamber (right panel). As a negative control, the primary antibody was eliminated from the staining procedure (left panel). Tissues were washed extensively with PBS/0.025% Tween-20 (Product # 28314) and endogenous peroxidase activity quenched with Peroxidase Suppressor (Product # 35000) for 30 minutes at room temperature. Detection was performed using an HRP-conjugated goat anti-rat IgG-HRP secondary antibody (Product # 31470) at a dilution of 1:500 followed by colorimetric detection using Metal Enhanced DAB Substrate Kit (Product # 34065). Tissues were counterstained with hematoxylin and prepped for mouting. Images were taken on a Zeiss Axiovision microscope at 40X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1. RegDCs EXO protect encapsulated immunoregulatory cargo (TGFB1 and IL10) from proteolytic degradation . (A) Nano-tracking analysis to determine EXO number and size distribution in nm. (B) Transmission electron microscopy (TEM) to visualize EXO shape and immuno-gold TEM to detect EXO marker tetraspanin CD63, showing unstained (left) and positive staining (right)(arrows). (C) Western blotting to detect other EXO-related markers including CD81, TSG101, GRP94 and B-actin in donor DCs and EXO (left) and anti/pro-inflammatory cytokines including TGFB1, IL10, IL6, IL1B and TNF and the costimulatory molecule CD86 as well as EXO associated proteins ALIX and TSG101 in DCs EXO subsets (right). (D) TGFB1 and IL10 content of lysed EXO and in regDCs EXO supernatant by ELISA. (E) TGFB1 and IL10 content of non-lysed EXO (transmembrane domain) by ELISA. (F) Immunogold TEM to detect luminal and transmembrane TGFB1 in regDCs EXO. (G) regDCs EXO (left) or equivalent concentration of free TGFB1 and IL10 (right) were treated with trypsin (upper panel) or proteinase-K (lower panel) and incubated in control buffer (1hour at 37degC) and analysed by western blotting to detect the levels of TGFB1, IL10 and exosomal markers TSG101 (upper panel) and CD81 (lower panel).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 5 Increased intracellular damages after AUY-VE. A673 cells were treated with 45 nM AUY922 +- 2 uM VE821 for 24 h. a Intracellular structures were analyzed by transmission electron microscopy (TEM). Red arrows indicate lipid-filled vesicles; yellow arrows indicate lysosomes. b Golgi networks were analyzed by fluorescence microscopy of GM130. Red arrows indicate dispersed Golgi structures. c Endoplasmic reticulum (ER) networks were analyzed by fluorescence microscopy of GRP94. Red arrows indicate vesicular ER structures. A673 cells were treated with 1-2 ug/ml of tunicamycin for 24 h. e ER networks were analyzed by fluorescence microscopy of GRP94. c - e DAPI was used to stain cell nuclei. A shows one representative experiments; B-E are representative for at least two independent experiments