Recombinant Anti-CD44 antibody [EPR18668] KO Tested (ab189524)

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Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR18668] to CD44
Suitable for: ICC/IF, Flow Cyt, WB, IHC-P, IP
Knockout validated
Reacts with: Mouse, Rat, Human

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Primary

Protein

Secondary

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Primary - Rabbit Anti-CD44 antibody [EPR18668] (ab189524)
ICC/IF, Flow Cyt, WB, IHC-P, IP

Protein - Recombinant human CD44 protein (Active) (ab173996)
ELISA, SDS-PAGE

Secondary - Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
IHC-P, WB, ELISA, IP

View more associated products

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
Positive control
- WB: Human fetal brain, fetal heart, fetal kidney and fetal spleen lysates; Human thymus and skin lysates; HeLa, 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
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

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

pH: 7.2
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
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant human CD44 protein (Active) (ab173996)
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
- CDW44 antigen 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
- HUTCHI antibody
- Hyaluronate receptor antibody
- IN antibody
- INLU-related p80 Glycoprotein antibody
- MC56 antibody
- MDU2 antibody
- MDU3 antibody
- MGC10468 antibody
- MIC4 antibody
- MUTCH I antibody
- MUTCH1 antibody
- PGP-1 antibody
- PGP-I antibody
- PGP1 antibody
- Phagocytic glycoprotein 1 antibody
- Phagocytic glycoprotein I antibody
- Soluble CD44 antibody
see all

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.
Western blot - Anti-CD44 antibody [EPR18668] (ab189524)

All lanes : Anti-CD44 antibody [EPR18668] (ab189524) at 1/1000 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: 80 kDa
why is the actual band size different from the predicted?

Lanes 1 - 4: Merged signal (red and green). Green - ab189524 observed at 80 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

ab189524 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 ab189524 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 1000 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.
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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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.
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.
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 Detection Reagent (HRP) (ab131366), was used for detection 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 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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

Click here to view the general protocols

Datasheet

Publishing research using ab189524? Please let us know so that we can cite the reference in this datasheet.

ab189524 has been referenced in 7 publications.

Zhang C & Wang Y Metformin attenuates cells stemness and epithelial-mesenchymal transition in colorectal cancer cells by inhibiting the Wnt3a/ß-catenin pathway. Mol Med Rep 19:1203-1209 (2019). PubMed: 30569135
Zhou ZQ et al. Adipose extracellular matrix promotes skin wound healing by inducing the differentiation of adipose-derived stem cells into fibroblasts. Int J Mol Med 43:890-900 (2019). PubMed: 30535488
Fan W et al. Identification of CD206 as a potential biomarker of cancer stem-like cells and therapeutic agent in liver cancer. Oncol Lett 18:3218-3226 (2019). PubMed: 31452799
Scalzone A et al. The interplay between chondrocyte spheroids and mesenchymal stem cells boosts cartilage regeneration within a 3D natural-based hydrogel. Sci Rep 9:14630 (2019). PubMed: 31601910
Takeuchi R et al. Exosomes from conditioned media of bone marrow-derived mesenchymal stem cells promote bone regeneration by enhancing angiogenesis. PLoS One 14:e0225472 (2019). PubMed: 31751396
Liu Q et al. Upregulation of musashi1 increases malignancy of hepatocellular carcinoma via the Wnt/ß-catenin signaling pathway and predicts a poor prognosis. BMC Gastroenterol 19:230 (2019). PubMed: 31888604
Yang Z et al. Cholesterol inhibits hepatocellular carcinoma invasion and metastasis by promoting CD44 localization in lipid rafts. Cancer Lett 429:66-77 (2018). IHC-P, ICC/IF . PubMed: 29746928

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Key features and details

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Recombinant Protein

Related Products

Applications

Target

Function

Tissue specificity

Sequence similarities

Domain

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Certificate of Compliance

References (7)

Customer reviews and Q&As

Post-translational
modifications