Recombinant Anti-CD44 antibody [EPR1013Y] - BSA and Azide free (ab216647)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR1013Y] to CD44 - BSA and Azide free
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-CD44 antibody [EPR1013Y] - BSA and Azide free
See all CD44 primary antibodies -
Description
Rabbit monoclonal [EPR1013Y] to CD44 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-Pmore details
Unsuitable for: Flow Cyt,ICC/IF or IP -
Species reactivity
Reacts with: Human
Does not react with: Mouse, Rat -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: MDA-MB-231, TF-1, HeLa and A549 cell lysates. IHC-P: Pancreatic and cervival cancer, breast and thyroid gland carcinoma, glioma, tonsil and skin tissues.
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General notes
ab216647 is the carrier-free version of ab51037.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Dissociation constant (KD)
KD = 3.76 x 10 -10 M Learn more about KD -
Storage buffer
pH: 7.20
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR1013Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- HRP Anti-CD44 antibody [EPR1013Y] (ab194989)
- Anti-CD44 antibody [EPR1013Y] - Low endotoxin, Azide free (ab246690)
- Alexa Fluor® 488 Anti-CD44 antibody [EPR1013Y] (ab311052)
- Alexa Fluor® 647 Anti-CD44 antibody [EPR1013Y] (ab311178)
- Alexa Fluor® 594 Anti-CD44 antibody [EPR1013Y] (ab311833)
- Alexa Fluor® 568 Anti-CD44 antibody [EPR1013Y] (ab313116)
- Alexa Fluor® 555 Anti-CD44 antibody [EPR1013Y] (ab313309)
- Anti-CD44 antibody [EPR1013Y] (ab51037)
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab216647 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
Use at an assay dependent concentration. Detects a band of approximately 82 kDa (predicted molecular weight: 82 kDa).
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Notes |
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WB
Use at an assay dependent concentration. Detects a band of approximately 82 kDa (predicted molecular weight: 82 kDa). |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Target
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Function
Receptor for hyaluronic acid (HA). Mediates cell-cell and cell-matrix interactions through its affinity for HA, and possibly also through its affinity for other ligands such as osteopontin, collagens, and matrix metalloproteinases (MMPs). Adhesion with HA plays an important role in cell migration, tumor growth and progression. Also involved in lymphocyte activation, recirculation and homing, and in hematopoiesis. Altered expression or dysfunction causes numerous pathogenic phenotypes. Great protein heterogeneity due to numerous alternative splicing and post-translational modification events. -
Tissue specificity
Isoform 10 (epithelial isoform) is expressed by cells of epithelium and highly expressed by carcinomas. Expression is repressed in neuroblastoma cells. -
Sequence similarities
Contains 1 Link domain. -
Domain
The lectin-like LINK domain is responsible for hyaluronan binding. -
Post-translational
modificationsProteolytically cleaved in the extracellular matrix by specific proteinases (possibly MMPs) in several cell lines and tumors.
N-glycosylated.
O-glycosylated; contains more-or-less-sulfated chondroitin sulfate glycans, whose number may affect the accessibility of specific proteinases to their cleavage site(s).
Phosphorylated; activation of PKC results in the dephosphorylation of Ser-706 (constitutive phosphorylation site), and the phosphorylation of Ser-672. -
Cellular localization
Membrane. - Information by UniProt
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Database links
- Entrez Gene: 960 Human
- Omim: 107269 Human
- SwissProt: P16070 Human
- Unigene: 502328 Human
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Alternative names
- LHR antibody
- BA-1 antibody
- CD 44 antibody
see all
Images
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All lanes : Anti-CD44 antibody [EPR1013Y] (ab51037) at 1/5000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : CD44 knockout HeLa cell lysate
Lane 3 : A549 cell lysate
Lane 4 : LNCaP cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 82 kDa
Observed band size: 75-80 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-CD44 antibody [EPR1013Y] staining at 1/5000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab51037 was shown to bind specifically to CD44. A band was observed at 75-80 kDa in wild-type HeLa cell lysates with no signal observed at this size in CD44 knockout cell line ab262515 (knockout cell lysate ab263938). To generate this image, wild-type and CD44 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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All lanes : Anti-CD44 antibody [EPR1013Y] (ab51037) at 1/5000 dilution
Lane 1 : Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : CD44 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 3 : A549 (Human lung carcinoma cell line) whole cell lysate
Lane 4 : LNCaP (Human prostate cancer cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 82 kDa
Observed band size: 80 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation (ab51037).
Lanes 1 - 4: Merged signal (red and green). Green - ab51037 observed at 80 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab51037 was shown to react with CD44 in wild-type HeLa cells in western blot. Loss of signal was observed when CD44 knockout sample was used. Wild-type HeLa and CD44 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab51037 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 5000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR1013Y] - BSA and Azide free (ab216647)
Immunohistochemical analysis of paraffin-embedded human skin tissue labeling CD44 with ab51037 at 1/100 dilution followed by goat anti-rabbit IgG H&L (HRP) (ab97051, 1/500). The sample was counter stained with hematoxylin.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab51037).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR1013Y] - BSA and Azide free (ab216647)
Immunohistochemical analysis of paraffin-embedded human pancreatic cancer tissue labeling CD44 with ab51037 at 1/100 dilution followed by goat anti-rabbit IgG H&L (HRP) (ab97051, 1/500). The sample was counter stained with hematoxylin.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab51037).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR1013Y] - BSA and Azide free (ab216647)
Immunohistochemical analysis of paraffin-embedded human cervical cancer tissue labeling CD44 with ab51037 at 1/100 dilution followed by goat anti-rabbit IgG H&L (HRP) (ab97051, 1/500). The sample was counter stained with hematoxylin.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab51037).
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Equilibrium disassociation constant (KD)
Learn more about KD
Click here to learn more about KDThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab51037).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR1013Y] - BSA and Azide free (ab216647)
This IHC data was generated using the same anti-CD44 antibody clone, EPR1013Y, in a different buffer formulation (cat# ab51037).
ab51037 (1:100) showing positive staining in human Breast carcinoma tissue.
Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR1013Y] - BSA and Azide free (ab216647)
This IHC data was generated using the same anti-CD44 antibody clone, EPR1013Y, in a different buffer formulation (cat# ab51037).
Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CD44 with ab51037 at 1/100 dilution followed by goat anti-rabbit IgG H&L (HRP) (ab97051, 1/500). The sample was counter stained with hematoxylin.
Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (37)
ab216647 has been referenced in 37 publications.
- Johnson BE et al. An omic and multidimensional spatial atlas from serial biopsies of an evolving metastatic breast cancer. Cell Rep Med 3:100525 (2022). PubMed: 35243422
- Burlingame EA et al. Toward reproducible, scalable, and robust data analysis across multiplex tissue imaging platforms. Cell Rep Methods 1:N/A (2021). PubMed: 34485971
- Li Y et al. Plantamajoside modulates the proliferation, stemness, and apoptosis of lung carcinoma via restraining p38MAPK and AKT phosphorylation. Transl Cancer Res 9:3828-3841 (2020). PubMed: 35117750
- Zhang L et al. CircAGFG1 drives metastasis and stemness in colorectal cancer by modulating YY1/CTNNB1. Cell Death Dis 11:542 (2020). PubMed: 32681092
- Gao Y et al. LINC00311 promotes cancer stem-like properties by targeting miR-330-5p/TLR4 pathway in human papillary thyroid cancer. Cancer Med 9:1515-1528 (2020). PubMed: 31894666