Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Rat, Naked mole-rat, Transgenic Mouse

Applications

Validated:

Knockout Validated, Immunohistochemistry, Western Blot, Flow Cytometry, Simple Western, Immunoprecipitation, CyTOF-ready

Cited:

Immunohistochemistry, Western Blot, Neutralization, Flow Cytometry, Immunocytochemistry, Simple Western, Immunoprecipitation, Cell Culture, Surface Plasmon Resonance

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2A Clone # 2C5
View all CD44 Primary Antibodies »

Product Specifications

Immunogen

Recombinant human CD44v3-10 (includes the invariant N-terminal exons and CD44v3-10 exons)

Specificity

Detects human CD44s on a panel of CD44 transfected COS cells by flow cytometry (Fox, S.B. et al. (1994) Cancer Res. 54:4539). This antibody recognizes an epitope in the invariant N-terminal region of all CD44 protein isoforms.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2A

Scientific Data Images for Human CD44s Pan Specific Antibody

Detection of Human CD44 antibody by Western Blot.

Detection of Human CD44 by Western Blot.

Western blot shows lysates of PC-3 human prostate cancer cell line and human tonsil tissue. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human CD44 s Pan Specific Monoclonal Antibody (Catalog # BBA10) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for CD44 at approximately 90 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

CD44 antibody in Human Lymphoma by Immunohistochemistry (IHC-P).

CD44 in Human Lymphoma.

CD44 was detected in immersion fixed paraffin-embedded sections of human lymphoma using Mouse Anti-Human CD44 s Pan Specific Monoclonal Antibody (Catalog # BBA10) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to plasma membrane. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Human CD44 antibody by Simple WesternTM.

Detection of Human CD44 by Simple WesternTM.

Simple Western lane view shows lysates of human tonsil tissue, loaded at 0.2 mg/mL. A specific band was detected for CD44 at approximately 156 kDa (as indicated) using 20 µg/mL of Mouse Anti-Human CD44 s Pan Specific Monoclonal Antibody (Catalog # BBA10). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Western Blot Shows CD44 Specificity Using Knockout Cell Line.

Western Blot Shows CD44 Specificity Using Knockout Cell Line.

Western blot shows lysates of MDA‑MB‑231 human breast cancer parental cell line and CD44 knockout MDA-MB-231 cell line (KO). Nitrocellulose membrane was probed with Mouse Anti-Human CD44 s Pan Specific Monoclonal Antibody (Catalog # BBA10) followed by HRP-conjugated secondary antibody. A specific band was detected for CD44 at approximately 81.5 kDa (as indicated) in the parental MDA-MB-231 cell line, but is not detectable in knockout MDA-MB-231 cell line. Primary antibody concentration used: 2.5 µg/mL. The Ponceau stained transfer of the blot is shown. This experiment was conducted under reducing conditions. Image, protocol, and testing courtesy of YCharOS Inc. See ycharos.com for additional details.
Detection of CD44 by Immunoprecipitation.

Detection of CD44 by Immunoprecipitation.

MDA‑MB‑231 human breast cancer cell line lysates were prepared and immunoprecipitation was performed using 2.0 μg of Mouse Anti-Human CD44 s Pan Specific Monoclonal Antibody (Catalog # BBA10) pre-coupled to Dynabeads Protein G. Immunoprecipitated CD44 was detected in Western Blot with a rabbit CD44 antibody used at 1/3000. The Ponceau stained transfer of the blot is shown. SM=4% starting material; UB=4% unbound fraction; IP=immunoprecipitate; HC=antibody heavy chain. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).
CD44 Specificity is Shown by Immunocytochemistry in Knockout Cell Line.

CD44 Specificity is Shown by Immunocytochemistry in Knockout Cell Line.

MDA‑MB‑231 human breast cancer parental cell line WT and CD44 MDA-MB-231 KO cells were labelled with a green or a far-red fluorescent dye, respectively. Cells were stained with Mouse Anti-Human CD44 s Pan Specific Monoclonal Antibody (Catalog # BBA10) followed by incubation with an Alexa-fluor 555 conjugated secondary antibody (upper panel). DAPI-only counterstained cells shown on a lower panel. Acquisition of the blue (nucleus-DAPI), green (identification of WT cells), red (antibody staining) and far-red (identification of KO cells) channels was performed. Representative images of the blue and red (grayscale) channels are shown. WT and KO cells are outlined with green and magenta dashed line, respectively. Primary antibody concentration used: 2 μg/mL. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).
Detection of CD44 by Flow Cytometry.

Detection of CD44 by Flow Cytometry.

MDA‑MB‑231 human breast cancer parental cell line WT and CD44 KO cells were labelled with a green or violet, fluorescent dye, respectively. WT and KO cells were mixed in a 1:1 ratio, fixed in 4% PFA and permeabilized in 0.1% saponin. 400,000 cells were stained with Mouse Anti-Human CD44 s Pan Specific Monoclonal Antibody (Catalog # BBA10) and a secondary antibody. Antibody staining was quantified, and representative images showing the staining intensity in the KO population (pink histogram, dashed line) compared to the WT cells (green histogram, solid line) are presented. Histograms with dotted lines represent secondary antibody-only controls in both WT and KO cells. Primary antibody concentration: 1 μg/mL. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).
Detection of Human CD44 by Immunocytochemistry/Immunofluorescence

Detection of Human CD44 by Immunocytochemistry/Immunofluorescence

Differential localization of Kv1.3 and KCa3.1 in migrating T cells.A. Distribution of Kv1.3 and KCa3.1 at the uropod. T cells were transiently transfected with either YFP-KCa3.1 or GFP-Kv1.3 (green) and stained with anti-CD44 antibody (uropod; red) without permeabilization. Yellow areas in the merge images indicate colocalization. Scale bar  = 5 µm. B. Distribution of Kv1.3 and KCa3.1 at the leading-edge. T cells, transfected with either YFP-KCa3.1 or GFP-Kv1.3 (green) and stained with anti-CXCR-4 antibody (leading-edge; red) without permeabilization, were analyzed by confocal microscopy. Colocalization between the two proteins is indicated by yellow areas in the merge images. Scale bar  = 5 µm. C. Correlation coefficients for KCa3.1 and Kv1.3 localization in the uropod (U) and leading-edge (L). The data are the average of n = 15 cells for KCa3.1 at the U and n = 8 at the L, and n = 16 for Kv1.3 at the U and n = 11 at the U from 2 healthy individuals. Statistical significance was established by one way ANOVA. D. Localization of native Kv1.3 in the leading-edge. T cells from one healthy individual were fixed and stained with extracellular anti-Kv1.3 antibody (green) together with antibodies either against CD44 (red; left) or CXCR-4 (red, right). Yellow colors in the merge images indicate strong correlation. Scale bar  = 5 µm. E. Average Correlation coefficients of native Kv1.3 with the leading-edge (L) (n = 9) and the uropod (U) markers (n = 11). Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0043859), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human CD44 by Immunocytochemistry/Immunofluorescence

Detection of Human CD44 by Immunocytochemistry/Immunofluorescence

Differential localization of Kv1.3 and KCa3.1 in migrating T cells.A. Distribution of Kv1.3 and KCa3.1 at the uropod. T cells were transiently transfected with either YFP-KCa3.1 or GFP-Kv1.3 (green) and stained with anti-CD44 antibody (uropod; red) without permeabilization. Yellow areas in the merge images indicate colocalization. Scale bar  = 5 µm. B. Distribution of Kv1.3 and KCa3.1 at the leading-edge. T cells, transfected with either YFP-KCa3.1 or GFP-Kv1.3 (green) and stained with anti-CXCR-4 antibody (leading-edge; red) without permeabilization, were analyzed by confocal microscopy. Colocalization between the two proteins is indicated by yellow areas in the merge images. Scale bar  = 5 µm. C. Correlation coefficients for KCa3.1 and Kv1.3 localization in the uropod (U) and leading-edge (L). The data are the average of n = 15 cells for KCa3.1 at the U and n = 8 at the L, and n = 16 for Kv1.3 at the U and n = 11 at the U from 2 healthy individuals. Statistical significance was established by one way ANOVA. D. Localization of native Kv1.3 in the leading-edge. T cells from one healthy individual were fixed and stained with extracellular anti-Kv1.3 antibody (green) together with antibodies either against CD44 (red; left) or CXCR-4 (red, right). Yellow colors in the merge images indicate strong correlation. Scale bar  = 5 µm. E. Average Correlation coefficients of native Kv1.3 with the leading-edge (L) (n = 9) and the uropod (U) markers (n = 11). Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0043859), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of CD44s Pan Specific by Western Blot

Detection of CD44s Pan Specific by Western Blot

A-B Expression of CD44st, CD44s, CD44v6 mRNA and miR-138-5p in breast cancer cell lines. A Semi-quantitative PCR agarose electrophoresis plots of different cell lines B Quantitative PCR results of different cell lines Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36964570), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of CD44s Pan Specific by Western Blot

Detection of CD44s Pan Specific by Western Blot

A-B Expression of CD44st, CD44s, CD44v6 mRNA and miR-138-5p in breast cancer cell lines. A Semi-quantitative PCR agarose electrophoresis plots of different cell lines B Quantitative PCR results of different cell lines Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36964570), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human CD44s Pan Specific Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

2.5 µg/106 cells
Sample: Human whole blood monocytes & MDA-MB-231 human breast cancer cell line lysates

Immunohistochemistry

8-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human lymphoma

Immunoprecipitation

Fox, S.B. et al. (1994) Cancer Res. 54:4539.

Knockout Validated

CD44 is specifically detected in MDA-MB-231 parental cell line but is not detectable in knockout MDA-MB-231 cell line.

Simple Western

20 µg/mL
Sample: Human tonsil tissue

Western Blot

2 µg/mL
Sample: PC‑3 human prostate cancer cell line and human tonsil tissue

Reviewed Applications

Read 14 reviews rated 4.6 using BBA10 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from ascites

Reconstitution

Sterile PBS to a final concentration of 0.5 mg/mL.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: CD44

CD44 is a ubiquitously expressed protein that is the major receptor for hyaluronan and exerts control over cell growth and migration (1-3). Human CD44 has a 20 amino acid (aa) signal sequence, an extracellular domain (ECD) with a 100 aa hyaluronan-binding disulfide-stabilized link region and a 325-530 aa stem region, a 21 aa transmembrane domain, and a 72 aa cytoplasmic domain. Within the stem, ten variably spliced exons (v1-10, exons 6-15) produce multiple protein isoforms (1‑3). The standard or hematopoietic form, CD44s, does not include the variable segments (1‑3). Cancer aggressiveness and T cell activation have been correlated with expression of specific isoforms (1, 3). With variable N- and O-glycosylation and splicing within the stalk, CD44 can range from 80-200 kDa (1). Within the N‑terminal invariant portion of the ECD (aa 21-220), human CD44 shares 76%, 76%, 86%, 83%, and 79% identity with corresponding mouse, rat, equine, canine, and bovine CD44, respectively. The many reported functions of CD44 fall within three categories (1). First, CD44 binds hyaluronan and other ligands within the extracellular matrix and can function as a “platform” for growth factors and metalloproteinases. Second, CD44 can function as a co-receptor that modifies activity of receptors including MET and the ERBB family of tyrosine kinases. Third, the CD44 intracellular domain links the plasma membrane to the actin cytoskeleton via the ERM proteins, ezrin, radixin and moesin. CD44 can be synthesized in a soluble form (4) or may be cleaved at multiple sites by either membrane-type matrix metalloproteinases, or ADAM proteases to produce soluble ectodomains (5, 6). The cellular portion may then undergo gamma secretase-dependent intramembrane cleavage to form an A beta -like transmembrane portion and a cytoplasmic signaling portion that affects gene expression (7, 8). These cleavage events are thought to promote metastasis by enhancing tumor cell motility and growth (1, 5).

References

  1. Ponta, H. et al. (2003) Nat. Rev. Mol. Cell Biol. 4:33.
  2. Screaton, G.R. et al. (1992) Proc. Natl. Acad. Sci. USA 89:12160.
  3. Lynch, K.W. (2004) Nat. Rev. Immunol. 4:931.
  4. Yu, Q. and B.P. Toole (1996) J. Biol. Chem. 271:20603.
  5. Nagano, O. and H. Saya (2004) Cancer Sci. 95:930.
  6. Nakamura, H. et al. (2004) Cancer Res. 64:876.
  7. Murakami, D. et al. (2003) Oncogene 22:1511.
  8. Lammich, S. et al. (2002) J. Biol. Chem. 277:44754.

Alternate Names

CD44, ECMR-III, HCAM, HCELL, LHR, MDU2, MDU3, MIC4, MUTCH-I, Pgp1

Entrez Gene IDs

960 (Human); 12505 (Mouse); 25406 (Rat); 100126860 (Porcine)

Gene Symbol

CD44

Additional CD44 Products

Product Documents for Human CD44s Pan Specific Antibody

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Product Specific Notices for Human CD44s Pan Specific Antibody

For research use only

Citations for Human CD44s Pan Specific Antibody

Customer Reviews for Human CD44s Pan Specific Antibody (14)

4.6 out of 5
14 Customer Ratings
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Showing  1 - 5 of 14 reviews Showing All
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  • Human CD44s Pan Specific Antibody
    Name: Anonymous
    Application: Immunohistochemistry
    Sample Tested: Neuroendocrine tumor
    Species: Human
    Verified Customer | Posted 04/19/2022
    Human CD44s Pan Specific Antibody BBA10
  • Human CD44s Pan Specific Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: Prostate cells
    Species: Human
    Verified Customer | Posted 08/15/2021
    Human CD44s Pan Specific Antibody BBA10
  • Human CD44s Pan Specific Antibody
    Name: Anonymous
    Application: ELISA
    Sample Tested: Serum and Plasma
    Species: Human
    Verified Customer | Posted 06/12/2019
    Human CD44s Pan Specific Antibody BBA10
  • Human CD44s Pan Specific Antibody
    Name: Anonymous
    Application: ELISA
    Sample Tested: Serum and Plasma
    Species: Human
    Verified Customer | Posted 01/30/2019
    MAB7045 was used as the capture antibody along with BBA10 as the detection antibody. Recombinant human CD44 Fc Chimera (3660-CD) was used as the calibrator material. Human serum and plasma samples were diluted 1:5 and all were quantifiable. Parallelism looked good.
    Human CD44s Pan Specific Antibody BBA10
  • Human CD44s Pan Specific Antibody
    Name: Anonymous
    Application: IHC for UMUC
    Sample Tested: Umbilical vein endothelial cells
    Species: Human
    Verified Customer | Posted 01/19/2019
    IHC for UMUC with one control 2 condition
    Human CD44s Pan Specific Antibody BBA10
  • Human CD44s Pan Specific Antibody
    Name: Anonymous
    Sample Tested: HUVEC human umbilical vein endothelial cells
    Species: Human
    Verified Customer | Posted 01/18/2019
    Chondroitiase treated increase CD44 when co-culture with macrophage
    Human CD44s Pan Specific Antibody BBA10
  • Human CD44s Pan Specific Antibody
    Name: Anonymous
    Application: ELISA
    Sample Tested: EDTA Plasma
    Species: Human
    Verified Customer | Posted 11/08/2018
  • Human CD44s Pan Specific Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: A549 human lung carcinoma cell line
    Species: Human
    Verified Customer | Posted 06/28/2017
  • Human CD44s Pan Specific Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Cell Lysates
    Species: Human
    Verified Customer | Posted 12/04/2016
    Protein expression confirms sorting results. Antibody is sensitive and reliable across multiple cell lines.
    Human CD44s Pan Specific Antibody BBA10
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: See PMID: 21364036
    Species: Human
    Verified Customer | Posted 01/16/2015
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: See PMID: 21730156
    Species: Human
    Verified Customer | Posted 01/16/2015
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: See PMID: 23736004
    Species: Human
    Verified Customer | Posted 01/16/2015
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: See PMID: 23457460
    Species: Human
    Verified Customer | Posted 01/16/2015
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: See PMID: 24512624
    Species: Human
    Verified Customer | Posted 01/16/2015

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