Anti-Hexokinase 1 antibody [EPR10134(B)] - BSA and Azide free

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Hexokinase 1 antibody [EPR10134(B)] - BSA and Azide free (AB233837)

Fluorescence multiplex immunohistochemical analysis of paraffin-embedded Human kidney tissue. Panel A : Merged staining of anti-Hexokinase 1 (gray; Opal™690), anti-Angiotensin Converting Enzyme 1 (green; Opal™520) and anti-Tissue Factor (red; Opal™570) on human kidney. Panel B : Anti-Tissue Factor stained on renal glomeruli. Panel C : Anti-Angiotensin Converting Enzyme 1 stained on proximal tubules. Panel D : Anti-Hexokinase 1 stained on distal tubules. The section was incubated in three rounds of staining : in the order of ab150423, ab254222, and ab252918 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. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. Counterstained with DAPI.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Hexokinase 1 antibody [EPR10134(B)] - BSA and Azide free (AB233837)

This data was developed using ab150423, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling Hexokinase 1 with Purified ab150423 at 1 : 50 dilution (4.1 μg/ml). Cells were fixed in 100% Methanol and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1 : 200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor® 488,ab150077) was used as the secondary antibody at 1 : 1000 (2 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Hexokinase 1 antibody [EPR10134(B)] - BSA and Azide free (AB233837)

This data was developed using ab150423, the same antibody clone in a different buffer formulation. Intracellular Flow Cytometry analysis of K-562 (Human chronic myelogenous leukemia lymphoblast) cells labeling Hexokinase 1 with Purified ab150423 at 1/20 dilution (10 μg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488 ,ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hexokinase 1 antibody [EPR10134(B)] - BSA and Azide free (AB233837)

This data was developed using ab150423, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human kidney tissue sections labeling Hexokinase 1 with purified ab150423 at 1/50 dilution (4.14 µg/mL). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Hexokinase 1 antibody [EPR10134(B)] - BSA and Azide free (AB233837)

Fluorescence multiplex immunohistochemical analysis of the human kidney (Formalin/PFA-fixed paraffin-embedded sections). Panel A : merged staining of anti-Hexokinase 1 (ab150423, green; Opal™690), anti-Angiotensin Converting Enzyme 1 (ab254222, gray; Opal™520) and anti-Aquaporin 2 (ab199975, red; Opal™570) on human kidney. Panel B : anti-Aquaporin 2 stained on collecting tubules. Panel C : anti-Angiotensin Converting Enzyme 1 stained on proximal tubules. Panel D : anti-Hexokinase 1 stained on distal tubules and collecting tubules. Opal Polymer HRP Ms + Rb was used as a secondary antibody. The section was incubated in three rounds of staining : in the order of ab150423 at 1/250 dilution (4.224 μg/ml), ab254222 at 1/4000 dilution (0.141 μg/ml) and ab199975 at 1/4000 dilution (0.152 μg/ml) 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. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. DAPI (blue) was used as a nuclear counter stain. This data was developed using ab150423, the same antibody clone in a different buffer formulation.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hexokinase 1 antibody [EPR10134(B)] - BSA and Azide free (AB233837)

This data was developed using ab150423, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat kidney tissue sections labeling Hexokinase 1 with purified ab150423 at 1/50 dilution (4.14 µg/mL). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hexokinase 1 antibody [EPR10134(B)] - BSA and Azide free (AB233837)

This data was developed using ab150423, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse kidney tissue sections labeling Hexokinase 1 with purified ab150423 at 1/50 dilution (4.14 µg/mL). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-Hexokinase 1 antibody [EPR10134(B)] - BSA and Azide free (AB233837)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded rat kidney tissue staining NADPH oxidase 4 with ab319055 at a 1 : 500 (1.032 ug/ml) dilution, ab233837 anti-Hexokinase 1 used at 1 : 250 (4.36 ug/ml) dilution and ab254222 anti-Angiotensin Converting Enzyme 1 used at a 1 : 4000 (0.132 ug/ml) dilution.

Panel A : merged staining of anti-NADPH oxidase 4 (green; Opal™520), anti-Hexokinase 1 (magenta; Opal™690) and anti-Angiotensin Converting Enzyme 1 (yellow; Opal™570) on rat kidney.
Panel B : anti-NADPH oxidase 4 staining distal tubules in rat kidney.
Panel C : anti-Hexokinase 1 staining distal tubules in rat kidney.
Panel D : anti-Angiotensin Converting Enzyme 1 staining proximal tubules in rat kidney.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab319055, ab233837 and ab254222 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.

Nuclear counter stain with DAPI.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-Hexokinase 1 antibody [EPR10134(B)] - BSA and Azide free (AB233837)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse kidney tissue staining NADPH oxidase 4 with ab319055 at a 1 : 500 (1.032 ug/ml) dilution, ab233837 anti-Hexokinase 1 used at 1 : 250 (4.36 ug/ml) dilution and ab254222 anti-Angiotensin Converting Enzyme 1 used at a 1 : 4000 (0.132 ug/ml) dilution.

Panel A : merged staining of anti-NADPH oxidase 4 (green; Opal™520), anti-Hexokinase 1 (magenta; Opal™690) and anti-Angiotensin Converting Enzyme 1 (yellow; Opal™570) on mouse kidney.
Panel B : anti-NADPH oxidase 4 staining distal tubules in mouse kidney.
Panel C : anti-Hexokinase 1 staining distal tubules in mouse kidney.
Panel D : anti-Angiotensin Converting Enzyme 1 staining proximal tubules in mouse kidney.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab319055, ab233837 and ab254222 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.

Nuclear counter stain with DAPI.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• WB

Unknown

Western blot - Anti-Hexokinase 1 antibody [EPR10134(B)] - BSA and Azide free (AB233837)

All lanes:

Western blot - Anti-Hexokinase 1 antibody [EPR10134(B)] - Mitochondrial Outer Membrane Marker (<a href='/en-us/products/primary-antibodies/hexokinase-1-antibody-epr10134b-mitochondrial-outer-membrane-marker-ab150423'>ab150423</a>) at 1/1000 dilution

Lane 1:

Human brain lysate

Lane 2:

Mouse brain lysate

Lane 3:

Rat brain lysate

Lane 4:

HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate

Lane 5:

MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate

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

Predicted band size: 102 kDa

false

• WB

Lab

Western blot - Anti-Hexokinase 1 antibody [EPR10134(B)] - BSA and Azide free (AB233837)

This data was developed using the same antibody clone in a different buffer formulation (ab150423).

Lanes 1-4 : Merged signal (red and green). Green - ab150423 observed at 102 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab150423 Anti-Hexokinase 1 antibody [EPR10134(B)] - Mitochondrial was shown to specifically react with Hexokinase 1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267279 (knockout cell lysate ab257161) was used. Wild-type and Hexokinase 1 knockout samples were subjected to SDS-PAGE. ab150423 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 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Hexokinase 1 antibody [EPR10134(B)] - Mitochondrial Outer Membrane Marker (<a href='/en-us/products/primary-antibodies/hexokinase-1-antibody-epr10134b-mitochondrial-outer-membrane-marker-ab150423'>ab150423</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

HK1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human HK1 (Hexokinase 1) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-hk1-hexokinase-1-knockout-hek-293t-cell-line-ab267279'>ab267279</a>)

Lane 3:

MCF7 cell lysate at 20 µg

Lane 4:

HEPG2 cell lysate at 20 µg

Predicted band size: 102 kDa

Observed band size: 102 kDa

false