CD44 Antibody (8E2F3) - BSA Free

Immunocytochemistry/Immunofluorescence: CD44 Antibody (8E2F3) [NBP1-47386] - Analysis of paraffin-embedded human lung cancer tissues using anti-CD44 mAb (green), showing membrane localization. DRAQ5 fluorescent DNA dye (blue).

Immunohistochemistry-Paraffin: CD44 Antibody (8E2F3) [NBP1-47386] - Analysis of FFPE human breast carcinoma tissues using CD44 antibody (8E2F3). The signal was developed using DAB based detection and the sections were processed for counterstaining with hematoxylin. The antibody generated mainly a membrane staining representative of CD44 protein.

Flow Cytometry: CD44 Antibody (8E2F3) [NBP1-47386] - Analysis of HeLa cells using anti-CD44 mAb (right) and negative control (left).

Immunocytochemistry/Immunofluorescence: CD44 Antibody (8E2F3) [NBP1-47386] - HeLa cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-CD44 Antibody [8E2F3] conjugated to DyLight 550 (NBP1-47386R) at 5 ug/ml for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Western Blot: CD44 Antibody (8E2F3) - BSA Free [NBP1-47386] - Generation and characterization of immortalized primary keratinocytes (IPKs) from K14Cre.Ptpn2w/w and K14Cre.Ptpn2fl/fl mice. Western blot analysis of pSTAT3, STAT3, and CD44 (NBP1-47386) in primary keratinocytes and IPKs from both genotypes. The A431 cells were used as positive controls. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29955047/) licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: CD44 Antibody (8E2F3) [NBP1-47386] - HeLa cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with CD44 Antibody [8E2F3] conjugated to Alexa Fluor 647 (NBP1-47386AF647) at 5 ug/ml for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Flow Cytometry: CD44 Antibody (8E2F3) [NBP1-47386] - An intracellular stain was performed on HeLa cells with NBP1-47386AF488 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.

Immunocytochemistry/Immunofluorescence: CD44 Antibody (8E2F3) [NBP1-47386] - Analysis of methanol-fixed A431 (A), HeLa (B), PANC-1 (C) and EC (D) cells using anti-CD44 mAb (green), showing membrane localization. DRAQ5 fluorescent DNA dye (blue).

Immunocytochemistry/Immunofluorescence: CD44 Antibody (8E2F3) [NBP1-47386] - Analysis of PANC-1 cells using anti-CD44 mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). DRAQ5 fluorescent DNA dye (blue).

Flow (Intracellular): CD44 Antibody (8E2F3) [NBP1-47386] - An intracellular stain was performed on HeLa cells with NBP1-47386AF647 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

Immunoprecipitation: CD44 Antibody (8E2F3) - BSA Free [NBP1-47386] - HAP1 lysates were prepared and immunoprecipitation was performed using 1.0 ug of the CD44 Antibody (NBP1-47386) pre-coupled to either protein G or protein A Sepharose beads. Ability of the antibodies to capture CD44 antigen was first assessed by comparing the level of CD44 antigen from the starting material (SM) to its level remaining in the unbound fractions (UB). Anti-CD44 antigen at 1/2000 was used for each immunoblot. Immunoprecipitate for CD44 Antibody (NBP1-47386) that showed depleted CD44 antigen in the UB can be seen. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).

CD44 (8E2F3) was detected in immersion fixed A431 human skin carcinoma cell line using Mouse anti- CD44 (8E2F3) Protein-G purified Monoclonal Antibody conjugated to Biotin (Catalog # NBP1-47386B) at 2 µg/mL overnight at 4C. Cells were stained using Streptavidin conjugated to DyLight 550 (red) and counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.

CD44 (8E2F3) was detected in immersion fixed A431 human skin carcinoma cell line using Mouse anti-CD44 (8E2F3) Protein-G purified Monoclonal Antibody conjugated to DyLight 550 (Catalog # NBP1-47386R) (red) at 5 µg/mL overnight at 4C. Cells were counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.

U-251 MG human glioblastoma cell line was stained with Mouse anti-CD44 (8E2F3) Protein-G purified Monoclonal Antibody conjugated to Alexa Fluor® 647 (Catalog # NBP1-47386AF647, blue histogram) or matched control antibody (orange histogram).

CD44 (8E2F3) was detected in immersion fixed NIH3T3 Mouse fibroblast cell line using Mouse anti- CD44 (8E2F3) Protein-G purified Monoclonal Antibody conjugated to DyLight 550 (Catalog # NBP1-47386R) (red) at 2 µg/mL overnight at 4C. Cells were counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.

An intracellular stain was performed on A431 human skin carcinoma cell line using Mouse anti-CD44 (8E2F3) Protein-G purified Monoclonal Antibody conjugated to DyLight 550 (Catalog # NBP1-47386R, blue histogram) or matched control antibody (orange histogram) at 2.5 µg/mL for 30 minutes at RT.

CD44 (8E2F3) was detected in immersion fixed U-2 OS human osteosarcoma cell line using Mouse anti-CD44 (8E2F3) Protein-G purified Monoclonal Antibody conjugated to Biotin (Catalog # NBP1-47386B) at 5 µg/mL overnight at 4C. Cells were stained using Streptavidin conjugated to DyLight 550 (red) and counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.

MDA‑MB‑231 human breast cancer cell line lysates were prepared and immunoprecipitation was performed using 2.0 μg of CD44 Antibody (8E2F3) - BSA Free (Catalog # NBP1-47386) 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).

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 CD44 Antibody (8E2F3) - BSA Free (Catalog # NBP1-47386) 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 dilution used: 1/1000. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).

• Reactivity: Hu, Mu, Rb(-), Pm, Ca, Eq
• Applications: WB, ELISA, Flow, ICC/IF, IHC, IP, KO
• Clone: 8E2F3
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1.0 mg/ml

CD44 Antibody (8E2F3) - BSA Free Summary

• Immunogen: Purified recombinant fragment of human CD44 (628-699) expressed in E. coli. [Uniprot: P16070]
• Localization: Membrane; Single-pass type I membrane protein.
• Marker: Cell Membrane Marker
• Predicted Species: Baboon (94%), Equine (93%), Canine (93%). Backed by our 100% Guarantee.
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: CD44
• Purity: Ammonium sulfate precipitation

Applications/Dilutions

• Dilutions:
  • ELISA 1:10000
  • Flow (Intracellular)
  • Flow Cytometry 1:200-1:400
  • Immunocytochemistry/ Immunofluorescence 1:200-1:1000
  • Immunohistochemistry 1:200-1:1000
  • Immunohistochemistry-Paraffin 1:200-1:1000
  • Immunoprecipitation
  • Knockout Validated Knockout validated from YCharOS Inc. (YCharOS.com)
  • Western Blot
• Theoretical MW: 82 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Control:

  HeLa Whole Cell Lysate

Reactivity Notes

Not reactive to rabbit per customer review.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Ammonium sulfate precipitation

Alternate Names for CD44 Antibody (8E2F3) - BSA Free

• CD44 antigen
• CD44 molecule (Indian blood group)
• CD44
• CD44R
• CDw44
• cell surface glycoprotein CD44
• chondroitin sulfate proteoglycan 8
• CSPG8
• ECMR-III
• Epican
• Extracellular matrix receptor III
• GP90 lymphocyte homing/adhesion receptor
• HCAM
• HCELL
• hematopoietic cell E- and L-selectin ligand
• Heparan sulfate proteoglycan
• Hermes antigen
• homing function and Indian blood group system
• HUTCH-I
• Hyaluronate receptor
• IN
• LHR
• MC56
• MDU2
• MDU2CD44 antigen (homing function and Indian blood group system)
• MDU3
• MDU3CDW44
• MIC4
• MIC4MGC10468
• MUTCH-I
• Pgp1
• PGP-1
• PGP-I
• Phagocytic glycoprotein 1
• Phagocytic glycoprotein I

Background

CD44, also known as IN, LHRMIC4, CDW44, HCELL, is a cell-surface glycoprotein involved in cell-cell interactions, cell adhesion and migration. It is a receptor for hyaluronic acid (HA) and can also interact with other ligands, such as osteopontin, collagens, and matrix metalloproteinases (MMPs). This protein participates in a wide variety of cellular functions including lymphocyte activation, recirculation and homing, hematopoiesis, and tumor metastasis. Transcripts for this gene undergo complex alternative splicing that results in many functionally distinct isoforms, however, the full length nature of some of these variants has not been determined. Alternative splicing is the basis for the structural and functional diversity of this protein, and may be related to tumor metastasis.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for CD44 Antibody (NBP1-47386)(22)

Publications using NBP1-47386

• Zuo Q, Yang Y, Lyu Y, Yang C, Chen C, Salman S, Huang T, Wicks E, Jackson W, Datan E, Qin W, Semenza G Plexin-B3 expression stimulates MET signaling, breast cancer stem cell specification, and lung metastasis. Cell Rep, 2023-02-28;42(3):112164. 2023-02-28 [PMID: 36857181]
• Lubanska D, Alrashed S, Mason GT et al. Impairing proliferation of glioblastoma multiforme with CD44+ selective conjugated polymer nanoparticles Scientific reports 2022-07-15 [PMID: 35840697] (PAGE)
• Saxena N, Bhardwaj G, Jadhav S, et al. Pre-metastatic niche drives breast cancer invasion by modulating MSC homing and CAF differentiation bioRxiv 2021-01-15 (ICC/IF, Human)
• Marzban H, Sasani F Canine Mammary Gland Cancer Stem Cell and its Potential Role in Malignant Biologic Behavior Iranian Journal of Veterinary Medicine 2020-01-01 (WB, Canine)
• Ning Li, Qin Zhu, Yuhua Tian, Kyung Jin Ahn, Xin Wang, Zvi Cramer, Justine Jou, Ian W. Folkert, Pengfei Yu, Stephanie Adams-Tzivelekidis, Priyanka Sehgal, Najia N. Mahmoud, Cary B. Aarons, Robert E. Roses, Andrei Thomas-Tikhonenko, Emma E. Furth, Ben Z. Stanger, Anil Rustgi, Malay Haldar, Bryson W. Katona, Kai Tan, Christopher J. Lengner Mapping and modeling human colorectal carcinoma interactions with the tumor microenvironment Nature Communications 2023-11-30 [PMID: 38036590]
• S Vondra, AL Höbler, AI Lackner, J Raffetsede, ZN Mihalic, A Vogel, L Saleh, V Kunihs, P Haslinger, M Wahrmann, H Husslein, R Oberle, J Kargl, S Haider, P Latos, G Schabbauer, M Knöfler, J Ernerudh, J Pollheimer The human placenta shapes the phenotype of decidual macrophages Cell Reports, 2023-01-10;42(1):111977. 2023-01-10 [PMID: 36640334]
• Salman S, Meyers DJ, Wicks EE Et al. HIF inhibitor 32-134D eradicates murine hepatocellular carcinoma in combination with anti-PD1 therapy J Clin Invest 2022-05-02 [PMID: 35499076] (FLOW, Mouse)
  
  Details:
  Citation using the Alexa Fluor 488 version of this antibody.
• Li H, Chaitankar V, Zhu J et al. Olfactomedin 4 mediation of prostate stem/progenitor-like cell proliferation and differentiation via MYC Scientific reports 2020-12-14 [PMID: 33318499] (ICC/IF, IHC-P, Human)
• Chen PY, Qin L, Li G et al. Smooth Muscle Cell Reprogramming in Aortic Aneurysms Cell Stem Cell 2020-04-02 [PMID: 32243809] (Mouse)
• Tsai FJ, Lai MT, Cheng J et al. Novel K6-K14 keratin fusion enhances cancer stemness and aggressiveness in oral squamous cell carcinoma Oncogene 2019-03-13 [PMID: 30867567] (IF/IHC, Mouse)
• Hassanzadeh H, Matin MM, Naderi-Meshkin H et al. Using paracrine effects of Ad-MSCs on keratinocyte cultivation and fabrication of epidermal sheets for improving clinical applications Cell Tissue Bank 2018-08-13 [PMID: 30105667] (FLOW, Human)
• Baek M, kim M, Lim JS et al. Epidermal-specific deletion of TC-PTP promotes UVB-induced epidermal cell survival through the regulation of Flk-1/JNk signaling. Cell Death Dis. 2018-06-28 [PMID: 29955047] (WB, Mouse)
• Jeong YJ, Oh HK, Park SH, Bong JG. Association between inflammation and cancer stem cell phenotype in breast cancer. Nat Commun. [PMID: 29434947] (IHC-P, Human)
• Samanta D, Park Y, Ni X et al. Chemotherapy induces enrichment of CD47+/CD73+/PDL1+ immune evasive triple-negative breast cancer cells Proc. Natl. Acad. Sci. U.S.A. 2018-01-24 [PMID: 29367423] (FLOW, Mouse)
  
  Details:
  This citation used the Alexa Fluor 488 form of this antbody
• Lee HC, Ling QD, Yu WC et al. Drug-resistant colon cancer cells produce high carcinoembryonic antigen and might not be cancer-initiating cells. Drug Des Devel Ther. 2013-01-01 [PMID: 23818760] (IF/IHC, Mouse)
• Papakonstantinou E, Klagas I, Karakiulakis G et al. Steroids and beta2-Agonists Regulate Hyaluronan Metabolism in Asthmatic Airway Smooth Muscle Cells Am J Respir Cell Mol Biol 2012-12-01 [PMID: 22865625]
• Du W, Wright BM, Li X, Finke J, Rini BI, Zhou M, He H, Lal P, Welford SM. Tumor-derived macrophage migration inhibitory factor promotes an autocrine loop that enhances renal cell carcinoma. Oncogene. 2012-04-30 [PMID: 22543583] (WB, Human)
• Asare O The Role of TC-PTP-Mediated Suppression of Autophagy During Skin Carcinogenesis Thesis 2023-01-01
• Samanta D, Huang T, Shah R et al. BIRC2 Expression Impairs Anti-Cancer Immunity and Immunotherapy Efficacy Cell Rep [PMID: 32846130] (FLOW, Mouse)
  
  Details:
  Citation using the PE format of this antibody.
• Byun Js, Oh M, Lee S Et Al. The transcription factor PITX1 drives astrocyte differentiation by regulating the SOX9 gene J. Biol. Chem. 2020-08-05 [PMID: 32759168] (IF/IHC, Rat)
• Scimeca M, Urbano N, Bonfiglio R et al. Breast osteoblast-like cells: a new biomarker for the management of breast cancer. Br. J. Cancer. 2018-10-17 [PMID: 30327566] (IHC-P, Human)
• Chefetz I, Alvero AB, Holmberg JC et al. TLR2 enhances ovarian cancer stem cell self-renewal and promotes tumor repair and recurrence. Cell Cycle 2013-02-01 [PMID: 23324344] (WB, IHC-P, FLOW, Human)

Review for CD44 Antibody (NBP1-47386) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Corey . WB Human 07/12/2013

Summary

• Application: Western Blot
• Sample Tested: Human Brain Tissue
• Species: Human
• Lot: 130226

Bioinformatics

• Gene Symbol: CD44
• Entrez:
  • 960(Human)
• Uniprot:
  • P16070()