Recombinant Anti-CD168 antibody [EPR4055] KO Tested (ab108339)

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Western blot - Anti-CD168 antibody [EPR4055] (ab108339)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR4055] to CD168
Suitable for: WB, IP, IHC-P
Knockout validated
Reacts with: Human

Carrier Free

Product name

Anti-CD168 antibody [EPR4055]
See all CD168 primary antibodies
Description

Rabbit monoclonal [EPR4055] to CD168
Host species

Rabbit
Tested applications

Suitable for: WB, IP, IHC-Pmore details
Unsuitable for: ICC/IF
Species reactivity

Reacts with: Human
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- T47-D MCF-7, SKBR-3 and LnCaP cell lysate Paraffin-embedded human breast carcinoma tissue Paraffin-embedded human testis tissue IP: T-47D whole cell lysate.
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 -20°C. Stable for 12 months at -20°C.
Storage buffer

pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR4055
Isotype

IgG
Research areas

Alternative Versions
- Anti-CD168 antibody [EPR4055] - Low endotoxin, Azide free (ab229447)
- Anti-CD168 antibody [EPR4055] - BSA and Azide free (ab271875)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Positive Controls
Recombinant Protein
- Recombinant Human CD168 protein (ab125997)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab108339 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
WB		1/1000 - 1/10000. Predicted molecular weight: 84 kDa.
IP		1/10 - 1/100.
IHC-P		1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Notes
WB 1/1000 - 1/10000. Predicted molecular weight: 84 kDa.
IP 1/10 - 1/100.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Application notes

Is unsuitable for ICC/IF.

Function

Involved in cell motility. When hyaluronan binds to HMMR, the phosphorylation of a number of proteins, including the focal adhesion kinase occurs. May also be involved in cellular transformation and metastasis formation, and in regulating extracellular-regulated kinase (ERK) activity.
Tissue specificity

Expressed in breast cancer cell lines and in normal breast tissue.
Cellular localization

Cell surface. Cytoplasm.
Target information above from: UniProt accession O75330 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 3161 Human
- Omim: 600936 Human
- SwissProt: O75330 Human
- Unigene: 728200 Human
Alternative names
- CD168 antibody
- CD168 antigen antibody
- HMMR antibody
- HMMR_HUMAN antibody
- Hyaluronan mediated motility receptor antibody
- Hyaluronan-mediated motility receptor (RHAMM) antibody
- IHABP antibody
- Intracellular hyaluronic acid-binding protein antibody
- MGC119494 antibody
- MGC119495 antibody
- OTTHUMP00000196920 antibody
- Receptor for hyaluronan-mediated motility antibody
- RHAMM antibody
see all

Western blot - Anti-CD168 antibody [EPR4055] (ab108339)

Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: Empty knockout HAP1 whole cell lysate (20 µg)
Lane 3: CD168 whole cell lysate (20 µg)

Lanes 1 - 3: Merged signal (red and green). Green - ab108339 observed at 90 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab108339 was shown to specifically react with CD168 when CD168 knockout samples were used. Wild-type and Empty knockout samples were subjected to SDS-PAGE. Ab108339 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1000 dilution and 1/10000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour at room temperature before imaging.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD168 antibody [EPR4055] (ab108339)

ab108339, at 1/100, staining CD168 in paraffin-embedded human breast carcinoma tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunoprecipitation - Anti-CD168 antibody [EPR4055] (ab108339)

Purified ab108339 at 1/50 dilution (2µg) immunoprecipitating CD168 in T-47D whole cell lysate.
Lane 1 (input): T-47D (Human ductal breast epithelial tumor epithelial cell) whole cell lysate 10µg
Lane 2 (+): ab108339 + T-47D whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab108339 in T-47D whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 85 kDa
Western blot - Anti-CD168 antibody [EPR4055] (ab108339)

All lanes : Anti-CD168 antibody [EPR4055] (ab108339) at 1/1000 dilution

Lane 1 : T47-D cell lysate
Lane 2 : MCF-7 cell lysate
Lane 3 : SKBR-3 cell lysate
Lane 4 : LnCaP cell lysate

Lysates/proteins at 10 µg per lane.

Predicted band size: 84 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD168 antibody [EPR4055] (ab108339)

ab108339, at 1/100, staining CD168 in paraffin-embedded human testis tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD168 antibody [EPR4055] (ab108339)

ab108339 showing positive staining in Normal thymus tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD168 antibody [EPR4055] (ab108339)

ab108339 showing positive staining in Normal stomach tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD168 antibody [EPR4055] (ab108339)

ab108339 showing negative staining in Normal brain tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD168 antibody [EPR4055] (ab108339)

ab108339 showing positive staining in Colonic adenocarcinoma tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD168 antibody [EPR4055] (ab108339)

ab108339 showing negative staining in Normal liver tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD168 antibody [EPR4055] (ab108339)

ab108339 showing positive staining in Normal tonsil tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Anti-CD168 antibody [EPR4055] (ab108339)

Western blot protocols
Immunoprecipitation protocols
Immunohistochemistry protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab108339? Please let us know so that we can cite the reference in this datasheet.

ab108339 has been referenced in 4 publications.

Chen YT et al. Immunohistochemical analysis of RHAMM expression in normal and neoplastic human tissues: a cell cycle protein with distinctive expression in mitotic cells and testicular germ cells. Oncotarget 9:20941-20952 (2018). PubMed: 29765511
Wang K & Zhang T Prognostic significance of CD168 overexpression in colorectal cancer. Oncol Lett 12:2555-2559 (2016). IHC ; Human . PubMed: 27698827
Wang D et al. Expression of the receptor for hyaluronic acid mediated motility (RHAMM) is associated with poor prognosis and metastasis in non-small cell lung carcinoma. Oncotarget 7:39957-39969 (2016). WB, IHC . PubMed: 27220886
Picaud L et al. Evaluation of the effects of hyaluronic acid-carboxymethyl cellulose barrier on ovarian tumor progression. J Ovarian Res 7:40 (2014). WB ; Human . PubMed: 24739440

Customer reviews and Q&As

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1-6 of 6 Abreviews or Q&A

Question

Could you please provide me with the concentrations for items ab124729 (lot GR82264-8) and ab108339?

Abcam community

Verified customer

Asked on Dec 03 2013

Answer

For ab124729 lot GR82264-8, the concentration is 1.797 mg/ml.

The current lot we have for sale for ab108339 is lot GR106937-4 and that is 0.193 mg/ml.

Elena Valued Customer

Abcam Scientific Support

Answered on Dec 03 2013

Question

The example on the Abcam website did say that fixation with paraformaldehyde was used but the cells were NOT permeabilized. So does this mean that the antibody recognizes denatured and not native protein or is the protein on the cell surface accessible only after the fixation process ?

Abcam community

Verified customer

Asked on Jan 15 2013

Answer

There should not be concerns about permeabilizing cells because as soon as they are fixed with parafomraldehyde, they are essentially permeabilized. The CD168 antigen is a non-integral cell surface hyaluronan receptor that is readily accessable to the antibody.

If you have not already done so, you might try an overnight incubation at 4 degrees C with the primary.

Abcam Scientific Support

Answered on Jan 15 2013

Question

Rabbit Mab, ab 108339 (clone EPR4055) – Do you have a protocol for using this antibody for flow cytometry ? I have tried this antibody using standard staining protocol and do not see any staining. Do the cells need to be fixed first (see 2D6 antibody) ?

Abcam community

Verified customer

Asked on Jan 14 2013

Answer

A link below is our Flow Cytometry protocol, which we use routinely for in-house testing.
http://www.epitomics.com/products/support/fc
Our protocol is for intracellular staining and we always fix the cells with 2% paraformaldehyde.

Abcam Scientific Support

Answered on Jan 14 2013

Question

Thank you for the information.

It is really important forthe customerto know exactly which of the four known isoforms of RHAMM ab108339 recognizes.

In the datasheet of the other antibody (ab67003) it says that it recognizes all 4 epitopes.

The 4 accession numbers of RHAMM isoforms are as follows:

a) NM_001142556 NP_001136028.1

b) NM_012484 NP_036616

c) NM_012485 NP_0.6617

d) NM_001142557 NP_001136029

Kindly advise.

Thanks in advance for your assistance and reply.

Looking forward to meeting you next week (during the distributor meeting).

Kind Regards,

Abcam community

Verified customer

Asked on Sep 03 2012

Answer

Thank you for your email.
I can confirm that ab108339 (Anti-CD168 antibody) would recognize all 4 isoforms of CD168/RHAMM.
If you need any further assistance in the future, please do not hesitate to contact me.

Abcam Scientific Support

Answered on Sep 03 2012

Question

Please send IHC-P protocol

Abcam community

Verified customer

Asked on Jul 23 2012

Answer

Thank you for contacting Abcam earlier today.

Please find attached the IHC-P protocol for both ab108339 & ab124729.

If there is anything else I can help you with, please let me know.

Abcam Scientific Support

Answered on Jul 23 2012

Question

Customer kindly asked
- the immunogenic info of antibody.
- Whether cells needs to permeabilized and fixed in FACS

Abcam community

Verified customer

Asked on Jan 24 2012

Answer

Thank you for contacting us.
The target CD168 is a cell surface protein. There no need for cell permeabilization and fixation (do not wait longer than an hour). Cell fixatiion is advised if there is a wait longer than an hour between analysis. Avoid the fixation if cells need to be viable.
Please click to link to find more information;
https://www.abcam.com/index.html?pageconfig=resource&rid=11380
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Abcam Scientific Support

Answered on Jan 24 2012

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Key features and details

Related conjugates and formulations

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

Positive Controls

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (4)

Customer reviews and Q&As