CD44 Antibody

SKU: orb251516

Description

CD44 Antibody

Images & Validation

• Tested Applications: IF, IHC, WB
• Reactivity: Human, Rat
• Application Notes:
  Western blot, 0.1-0.5μg/ml, Human, Rat Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human Immunofluorescence, 2μg/ml, Human, Rat. Add 0.2ml of distilled water will yield a concentration of 500ug/ml

Key Properties

• Antibody Type: Primary Antibody
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: Rabbit IgG
• Immunogen: A synthetic peptide corresponding to a sequence at the N-terminus of human CD44, different from the related mouse and rat sequences by two amino acids.
• Molecular Weight: 82 kDa
• Purification: Immunogen affinity purified.
• Conjugation: Unconjugated

Storage & Handling

• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Form/Appearance: Lyophilized
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Disclaimer: For research use only

Alternative Names

CD44 antigen; CDw44; Epican; Extracellular matrix receptor III; ECMR-III; GP90 lymphocyte homing/adhesion receptor; HUTCH-I; Heparan sulfate proteoglycan; Hermes antigen; Hyaluronate receptor; Phagocytic glycoprotein 1; PGP-1; Phagocytic glycoprotein I; PGP-I; CD44; CD44; LHR, MDU2, MDU3, MIC4

IF analysis of CD44 using anti-CD44 antibody. CD44 was detected in paraffin-embedded section of human tonsil tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/mL rabbit anti-CD44 Antibody overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

IF analysis of CD44 using anti-CD44 antibody. CD44 was detected in paraffin-embedded section of rat lymphaden tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/mL rabbit anti-CD44 Antibody overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

IHC analysis of CD44 using anti-CD44 antibody. CD44 was detected in a paraffin-embedded section of Human Intestinal Cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml rabbit anti-CD44 Antibody overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.

Western blot analysis of CD44 using anti-CD44 antibody. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% milk in PBS/0.05% Tween-20 (5% milk/PBS/Tw) for 1.5 hour at RT. The membrane was incubated with rabbit anti-CD44 antibody at 1 ug/mL in 5% milk/PBS/0.05% Tween 20 overnight at 4℃, then washed with TBS-0.1% Tween 3 times with 5 minutes each and probed with a goat anti-rabbit antibody conjugated with HRP at 1:5000 in 5% milk/PBS/Tw at 4℃ for 12 hours. The signal is developed using an SuperSignal West Pico Chemiluminescent Substrate. A specific band was detected for EGFR at approximately 20 kDa. The expected band size for EGFR is at 81 kDa.

Western blot analysis of CD44 using anti-CD44 antibody. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions. Lane 1: human Hela whole cell lysates, Lane 2: human U87 whole cell lysates, Lane 3: rat PC-12 whole cell lysates, Lane 4: rat C6 whole cell lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-CD44 antigen affinity purified polyclonal antibody at 0.5 µg/mL overnight at 4°C, then washed with TBS-0.1% Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for CD44 at approximately 82 kDa. The expected band size for CD44 is at 82 kDa.

Quick Database Links

UniProt

P16070