Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-CD168 antibody [EPR4054] - Low endotoxin, Azide free (ab228482)

Overview

  • Product name

    Anti-CD168 antibody [EPR4054] - Low endotoxin, Azide free
    See all CD168 primary antibodies
  • Description

    Rabbit monoclonal [EPR4054] to CD168 - Low endotoxin, Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, IHC-Pmore details
    Unsuitable for: Flow Cyt or ICC
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide aa 1-100. The exact sequence is proprietary.

  • Positive control

    • Lysates of: MDA-MB-435, LnCaP, T47-D, RAW264.7, PC-12. Human testis tissue
  • General notes

    The formulation and the concentration of this product is compatible for metal-conjugation for mass cytometry (CyTOF®). 

    ab228482 is a PBS only buffer version of ab124729, containing no BSA or sodium azide, ideal for antibody labeling. Please refer to ab124729 for information on protocols, dilutions, and image data.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Properties

Applications

Our Abpromise guarantee covers the use of ab228482 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 Use at an assay dependent concentration. Predicted molecular weight: 84 kDa.

The mouse and rat recommendation is based on the WB results. This antibody may not be suitable for IHC with mouse or rat samples.

IP Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.

Antigen retrieval is recommended

  • Application notes
    Is unsuitable for Flow Cyt or ICC.
  • Target

    • 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.
    • Information by UniProt
    • Database links

    • 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

    Images

    • This WB data was generated using the same anti-CD168 antibody clone, EPR4054, in a different buffer formulation (cat# ab124729).

      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 - ab124729 observed at 90 kDa. Red - loading control, ab9484, observed at 37 kDa.

      ab124729 was shown to specifically react with CD168 when CD168 knockout samples were used. Wild-type and Empty knockout samples were subjected to SDS-PAGE.  Ab124729 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.

    • ab124729 (purified) at 1:100 dilution (2ug) immunoprecipitating CD168 in MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysate.

      Lane 1 (input): MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysate 10ug

      Lane 2 (+): ab124729 & MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysate

      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab124729 in MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysate

      For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.

      Blocking and diluting buffer: 5% NFDM/TBST.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124729).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human gastric carcinoma tissue sections labeling CD168 with purified  ab124729 at 1:250 dilution (7.7 μg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using EDTA Buffer, pH9.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124729).

    • Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124729).

    • This IHC data was generated using the same anti-CD168 antibody clone, EPR4054, in a different buffer formulation (cat# ab124729).

      Immunohistochemical analysis of paraffin-embedded human testis tissue using  unpurified ab124729 at 1/50 - 1/100 dilution

    References

    ab228482 has not yet been referenced specifically in any publications.

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