Immunohistochemical analysis of paraffin-embedded human skin tissue labeling CD44 with ab157107 at 1/2000 dilution followed by goat anti-rabbit IgG H&L (HRP) (ab97051, 1/500). The sample was counter stained with hematoxylin.

ab157107 staining CD44 in C6 cell membranes by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% PFA, permeabilised with 0.1% Triton-X and incubated with primary antibody (1/1000). An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (ab150077) was used as the secondary antibody at a dilution of 1/1000, shown in the top left hand panel. Additionally, ab7291 anti-tubulin and nuclear stain DAPI (blue) were used as counterstains as shown in the top right hand panel.

Negative control 1: Primary ab157107 and ab150120 Alexa Fluor^®594 goat anti-mouse secondary were both used at a dilution of 1/1000.

Negative control 2: ab7291 anti-tubulin and ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary were both used at a dilution of 1/1000.

All lanes : Anti-CD44 antibody (ab157107) at 1/10000 dilution

Lane 1 : MCF-7 (human breast adenocarcinoma epithelial) whole cell lysates
Lane 2 : Jurkat (human acute T cell leukaemia lymphocyte) whole cell lysates
Lane 3 : MDA-MB-231 (human breast adenocarcinoma epithelial) whole cell lysates
Lane 4 : HeLa (human cervix adenocarcinoma epithelial) whole cell lysates

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 81 kDa
Additional bands at: 81 kDa. We are unsure as to the identity of these extra bands.


Exposure time: 10 seconds

Blocking buffer: 5% NFDM/TBST

Diluting buffer: 5% NFDM/TBST

The expression of CD44 in MCF-7 is low (PMID: 25635866; PMID: 26005723). Jurkat does not express CD44 (PMID: 24127558).

All lanes : Anti-CD44 antibody (ab157107) at 1/10000 dilution

All lanes :

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Rabbit polyclonal to GNAT2 (ab97501) at 1/20000 dilution

Predicted band size: 81 kDa


Exposure time: 10 seconds

Blocking buffer: 5% NFDM/TBST

Diluting Buffer: 5% NFDM/TBST

All lanes : Anti-CD44 antibody (ab157107) at 1/10000 dilution

Lane 1 : NIH/3T3 (mouse embryo fibroblast) whole cell lysates
Lane 2 : Raw264.7 (mouse macrophage) whole cell lysates
Lane 3 : Mouse brain tissue lysates
Lane 4 : Mouse spleen tissue lysates

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 81 kDa
Additional bands at: 81 kDa. We are unsure as to the identity of these extra bands.


Exposure time: 1 minute

Blocking Buffer: 5% NFDM/TBST

Diluting Buffer: 5% NFDM/TBST

All lanes : Anti-CD44 antibody (ab157107) at 1/10000 dilution

Lane 1 : Human breast cancer tissue lysate
Lane 2 : Human lung tissue lysate
Lane 3 : Human spleen tissue lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 81 kDa
Additional bands at: 81 kDa. We are unsure as to the identity of these extra bands.


Exposure time: 3 minutes

Blocking buffer: 5% NFDM/TBST

Diluting buffer: 5% NFDM/TBST

 

All lanes : Anti-CD44 antibody (ab157107) at 0.1 µg/ml

Lane 1 : HeLa whole cell lysate at 50 µg
Lane 2 : HeLa whole cell lysate at 15 µg
Lane 3 : 293T whole cell lysate at 50 µg
Lane 4 : Jurkat whole cell lysate at 50 µg

Developed using the ECL technique.

Predicted band size: 81 kDa


Exposure time: 3 minutes

Detection of CD44 in Immunoprecipitates of HeLa whole cell lysates (1 mg for IP, 20% of IP loaded) using ab157107(Lane 1). For WB detection an ab157107 was used at 1 µg/ml. Lane 2 represents control IgG IP. Detection: Chemiluminescence with an exposure time of 30 seconds.

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CD44 with ab157107 at 1/2000 dilution followed by goat anti-rabbit IgG H&L (HRP) (ab97051, 1/500). The sample was counter stained with hematoxylin.

Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling CD44 with ab157107 at 1/2000 dilution followed by goat anti-rabbit IgG H&L (HRP) (ab97051, 1/500). The sample was counter stained with hematoxylin.

Immunohistochemical analysis of paraffin-embedded human pancreas cancer tissue labeling CD44 with ab157107 at 1/2000 dilution followed by goat anti-rabbit IgG H&L (HRP) (ab97051, 1/500). The sample was counter stained with hematoxylin.

Immunohistochemical analysis of paraffin-embedded human cervical cancer tissue labeling CD44 with ab157107 at 1/2000 dilution followed by goat anti-rabbit IgG H&L (HRP) (ab97051, 1/500). The sample was counter stained with hematoxylin.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian carcinoma tissue labelling CD44 with ab157107 at 1/1000 (1µg/ml). Detection: DAB.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon carcinoma tissue labelling CD44 with ab157107 at 1/1000 (1µg/ml). Detection: DAB.

ab157107 staining CD44 in MDA-MB-231 cell membranes by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% PFA and permeabilized with 0.1% Triton-X. Samples were incubated with primary antibody and an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (ab150077) was used as the secondary antibody, both at a dilution of 1/1000, shown in the top left hand panel. Additionally, ab7291 anti-tubulin and nuclear stain DAPI (blue) were used as counterstains as shown in the top right hand panel.

Negative control 1: Primary ab157107 and ab150120 Alexa Fluor^®594 goat anti-mouse secondary were both used at a dilution of 1/1000.

Negative control 2: ab7291 anti-tubulin and ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary were both used at a dilution of 1/1000.

ab157107 showing negative staining of CD44 in MCF-7 cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% PFA and permeabilized with 0.1% Triton-X. Samples were incubated with primary antibody and an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (ab150077) was used as the secondary antibody, both at a dilution of 1/1000, shown in the top left hand panel. Additionally, ab7291 anti-tubulin and nuclear stain DAPI (blue) were used as counterstains as shown in the top right hand panel.

Negative control 1: Primary ab157107 and ab150120 Alexa Fluor^®594 goat anti-mouse secondary were both used at a dilution of 1/1000.

Negative control 2: ab7291 anti-tubulin and ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary were both used at a dilution of 1/1000.

ab157107 showing weak staining of CD44 in NIH/3T3 cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 100% methanol and incubated with primary antibody (1/1000). An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (ab150077) was used as the secondary antibody at a dilution of 1/1000, shown in the top left hand panel. Additionally, ab7291 anti-tubulin and nuclear stain DAPI (blue) were used as counterstains as shown in the top right hand panel.

Negative control 1: Primary ab157107 and ab150120 Alexa Fluor^®594 goat anti-mouse secondary were both used at a dilution of 1/1000.

Negative control 2: ab7291 anti-tubulin and ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary were both used at a dilution of 1/1000.