Anti-CD44 antibody [EPR18668] Recombinant (ab189524)

Product name

Anti-CD44 antibody [EPR18668]
See all CD44 primary antibodies
Description

Rabbit monoclonal [EPR18668] to CD44
Host species

Rabbit
Tested applications

Suitable for: ICC/IF, Flow Cyt, WB, IHC-P, IPmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Recombinant fragment within Human CD44 aa 1-250. The exact sequence is proprietary.
Database link: P16070

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Positive control
- WB: Human fetal brain, fetal heart, fetal kidney and fetal spleen lysates; Human thymus and skin lysates; A549, U-87 MG, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Mouse brain and spleen lysates; Rat brain, heart, kidney and spleen lysates; IHC-P: Human breast, kidney, tonsil and breast cancer tissues; Mouse colon, stomach and spleen tissues; Rat stomach and spleen tissues; IP: A549 whole cell lysate. ICC/IF: MDA-MB-231 cell line. Flow Cytometry: MDA-MB-231 cell line.
General notes

Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab^® patents
This product is a recombinant rabbit monoclonal antibody.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR18668
Isotype

IgG
Research areas

Alternative Versions
- Anti-CD44 antibody [EPR18668] - BSA and Azide free (ab232556)
Compatible Secondaries
Related Products
- VeriBlot for IP Detection Reagent (HRP) (ab131366)
- Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

Our Abpromise guarantee covers the use of ab189524 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
ICC/IF		1/250.
Flow Cyt		1/40.
WB		1/1000. Detects a band of approximately 82 kDa (predicted molecular weight: 82 kDa).
IHC-P		1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP		1/30.

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
modifications

Proteolytically 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.
Target information above from: UniProt accession P16070 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 960 Human
- Entrez Gene: 12505 Mouse
- Entrez Gene: 25406 Rat
- Omim: 107269 Human
- SwissProt: P16070 Human
- SwissProt: P15379 Mouse
- SwissProt: P26051 Rat
- Unigene: 502328 Human
- Unigene: 423621 Mouse
- Unigene: 1120 Rat
see all
Alternative names
- LHR antibody
- BA-1 antibody
- CD 44 antibody
- CD44 antibody
- CD44 antigen antibody
- CD44 molecule (Indian blood group) antibody
- CD44 molecule antibody
- CD44_HUMAN antibody
- CDw44 antibody
- Cell surface glycoprotein CD44 antibody
- chondroitin sulfate proteoglycan 8 antibody
- CSPG8 antibody
- ECMR-III antibody
- Epican antibody
- Extracellular matrix receptor III antibody
- GP90 lymphocyte homing/adhesion receptor antibody
- HCELL antibody
- hematopoietic cell E- and L-selectin ligand antibody
- Heparan sulfate proteoglycan antibody
- Hermes antigen antibody
- homing function and Indian blood group system antibody
- HSA antibody
- HUTCH-I antibody
- HUTCH1 antibody
- Hyaluronate receptor antibody
- IN antibody
- INLU-related p80 Glycoprotein antibody
- MC56 antibody
- MDU2 antibody
- MDU3 antibody
- MGC10468 antibody
- MIC4 antibody
- MUTCH1 antibody
- PGP-1 antibody
- PGP-I antibody
- PGP1 antibody
- Phagocytic glycoprotein 1 antibody
- Phagocytic glycoprotein I antibody
- Soluble CD44 antibody
see all

Immunocytochemistry/ Immunofluorescence - Anti-CD44 antibody [EPR18668] (ab189524)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MDA-MB-231 (Human breast adenocarcinoma cell line) labeling CD44 with ab189524 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing membranous staining on MDA-MB-231 cell line.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with Mouse monoclonal [DM1A] to alpha Tubulin - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR18668] (ab189524)

Immunohistochemical analysis of paraffin-embedded human breast tissue labeling CD44 with ab189524 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and weak cytoplasmic staining on human breast [PMID: 20103682].

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR18668] (ab189524)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling CD44 with ab189524 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and weak cytoplasmic staining on human kidney.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Western blot - Anti-CD44 antibody [EPR18668] (ab189524)

All lanes : Anti-CD44 antibody [EPR18668] (ab189524) at 1/1000 dilution

Lane 1 : Human fetal brain lysate
Lane 2 : Human fetal heart lysate
Lane 3 : Human fetal kidney lysate
Lane 4 : Human fetal spleen lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/10000 dilution

Predicted band size: 82 kDa
Observed band size: 82 kDa

Exposure time: 5 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-CD44 antibody [EPR18668] (ab189524)

All lanes : Anti-CD44 antibody [EPR18668] (ab189524) at 1/2000 dilution

Lane 1 : Human thymus lysate
Lane 2 : Human skin lysate
Lane 3 : A549 (Human lung carcinoma cell line) whole cell lysate
Lane 4 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
Lanes 1-2 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/10000 dilution
Lanes 3-4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 82 kDa
Observed band size: 82 kDa

Exposure time: 1 second

Blocking/Dilution buffer: 5% NFDM/TBST.

Due to high sequence homology between CD44 isoforms, within the immunogen region, this antibody cross reacts with most isoforms of CD44. The staining pattern is consistent with reference (PMID 19507218).
Western blot - Anti-CD44 antibody [EPR18668] (ab189524)

Lanes 1-6 : Anti-CD44 antibody [EPR18668] (ab189524) at 1/1000 dilution
Lanes 7-10 : Anti-CD44 antibody [EPR18668] (ab189524) at 1/20000 dilution

Lane 1 : Mouse brain lysate at 10 µg
Lane 2 : Mouse spleen lysate at 10 µg
Lane 3 : Rat brain lysate at 10 µg
Lane 4 : Rat heart lysate at 10 µg
Lane 5 : Rat kidney lysate at 10 µg
Lane 6 : Rat spleen lysate at 10 µg
Lane 7 : C6 (Rat glial tumor cell line) whole cell lysate
Lane 8 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 9 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 10 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 82 kDa
Observed band size: 82 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1, 2, 3, 4, 5 and 6: 5 seconds; Lane 7, 8, 9 and 10: 1 second.
Flow Cytometry - Anti-CD44 antibody [EPR18668] (ab189524)

Flow cytometric analysis of 4% paraformaldehyde-fixed MDA-MB-231 (Human breast adenocarcinoma cell line) cell line labeling CD44 with ab189524 at 1/40 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.
Immunoprecipitation - Anti-CD44 antibody [EPR18668] (ab189524)

CD44v6 was immunoprecipitated from 1 mg of A549 (Human lung carcinoma cell line) whole cell lysate with ab189524 at 1/25 dilution. Western blot was performed from the immunoprecipitate using ab189524 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/10000 dilution.

Lane 1: A549 whole cell lysate 10 μg (Input).

Lane 2: ab189524 IP in A549 whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab189524 in A549 whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 1 second.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR18668] (ab189524)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CD44 with ab189524 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and weak cytoplasmic staining on human tonsil.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR18668] (ab189524)

Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling CD44 with ab189524 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and weak cytoplasmic staining on human breast cancer [PMID: 15867228].

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR18668] (ab189524)

Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling CD44 with ab189524 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and weak cytoplasmic staining on mouse colon.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR18668] (ab189524)

Immunohistochemical analysis of paraffin-embedded mouse stomach tissue labeling CD44 with ab189524 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and weak cytoplasmic staining on mouse stomach.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR18668] (ab189524)

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CD44 with ab189524 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and weak cytoplasmic staining on mouse spleen.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR18668] (ab189524)

Immunohistochemical analysis of paraffin-embedded rat stomach tissue labeling CD44 with ab189524 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous staining on rat stomach.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR18668] (ab189524)

Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling CD44 with ab189524 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and weak cytoplasmic staining on rat spleen.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Datasheet

Flow cytometry protocols
Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

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Anti-CD44 antibody [EPR18668] (ab189524)

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