Anti-HMG4 antibody [EP2839Y]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HMG4 antibody [EP2839Y] (AB75782)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human placenta tissue sections labeling HMG4 with Purified ab75782 at 1/100 dilution (1.13 μg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-HMG4 antibody [EP2839Y] (AB75782)

Immunocytochemistry analysis of 293T (Human embryonic kidney epithelial cell) cells labeling HMG4 with Purified ab75782 at 1/250 dilution (0.452 μg/ml). Cells were fixed in 4% Paraformaldehyde 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-HMG4 antibody [EP2839Y] (AB75782)

Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling HMG4 with Purified ab75782 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).

• IP

Lab

Immunoprecipitation - Anti-HMG4 antibody [EP2839Y] (AB75782)

Purified ab75782 at 1/20 dilution (0.5μg) immunoprecipitating HMG4 in HeLa whole cell lysate.

Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10μg
Lane 2 : ab75782 + HeLa whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab75782 in HeLa whole cell lysate.

VeriBlot for IP Detection Reagent (HRP)(ab131366) (1/1000 dilution) was used for Western blotting.

Blocking Buffer and concentration : 5% NFDM/TBST.

Observed band size : 25 kDa

All lanes:

Immunoprecipitation - Anti-HMG4 antibody [EP2839Y] (ab75782)

Predicted band size: 23 kDa

false

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HMG4 antibody [EP2839Y] (AB75782)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labeling HMG4 with Purified ab75782 at 1/100 dilution (1.13 μg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) 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-HMG4 antibody [EP2839Y] (AB75782)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat liver tissue sections labeling HMG4 with Purified ab75782 at 1/100 dilution (1.13 μg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• WB

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Western blot - Anti-HMG4 antibody [EP2839Y] (AB75782)

Blocking buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-HMG4 antibody [EP2839Y] (ab75782) at 1/5000 dilution

All lanes:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 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

Predicted band size: 23 kDa

Observed band size: 25 kDa

false

• WB

Unknown

Western blot - Anti-HMG4 antibody [EP2839Y] (AB75782)

Blocking buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-HMG4 antibody [EP2839Y] (ab75782) at 1/1000 dilution

Lane 1:

Human bladder lysate at 20 µg

Lane 2:

Human placenta lysate at 20 µg

Lane 3:

Mouse brain lysate at 20 µg

Lane 4:

Mouse bladder lysate at 20 µg

Lane 5:

Rat brain lysate at 20 µg

Lane 6:

Rat bladder lysate at 20 µg

Lane 7:

Rat placenta 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

Predicted band size: 23 kDa

Observed band size: 25 kDa

false