重组Anti-CD44抗体[Hermes-3] (ab254530)

ab254530 was shown to react with CD44 in wild-type HAP1 cells in Western blot with loss of signal observed in a CD44 knockout cell line. Wild-type HAP1 and CD44 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab254530 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with goat anti-mouse HRP secondary antibodies at 0.3ug/mL before imaging. These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

Blocking/Dilution buffer: 5% NFDM/TBST.

Immunohistochemical analysis of paraffin-embedded human skin tissue labeling CD44 with ab254530 at 0.107µg/ml, followed by ready to use secondary. Membranous staining on human skin is observed. The section was incubated with ab254530 for 5 mins at RT. The immunostaining staining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, followed by ready to use secondary.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

ab254530 was shown to react with CD44 in wild-type HAP1 cells in Immunocytochemistry with loss of signal observed in a CD44 knockout cell line. Wild-type and knockout cells were mixed and pelleted at a 1:1 ratio on coverslips. The cells were fixed with 4% paraformaldehyde (15 min) then permeabilized with 0.1% Triton X-100 (10min) and then blocked. The cells were then incubated with ab254530 at 1/500 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a secondary antibody to (Alexa Fluor^® 555) at 0.5 μg/ml. Acquisition of the green (wild-type), red (antibody staining) and far-red (knockout) channels was performed. Representative grayscale images of the red channel are shown. Wild-type and knockout cells are outlined with yellow and magenta dashed line, respectively. Schematic representation of the mosaic strategy used is shown on the bottom-right panel. Image was acquired with a Zeiss(LSM-880). These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

Immunohistochemical analysis of paraffin-embedded human bladder carcinoma tissue labeling CD44 with ab254530 at 0.107µg/ml, followed by ready to use secondary. Membranous staining on human bladder carcinoma is observed. The section was incubated with ab254530 for 5 mins at RT. The immunostaining staining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, followed by ready to use secondary.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

ELISA - Anti-CD44 antibody [Hermes-3] (ab254530).

Antigen: Human CD44.

ab254530 used at 100-1000µg/ml.

Secondary is an Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Mouse IgG (H+L) used at a 1/1000 dilution.