Recombinant
RabMAb

Recombinant Anti-CD68 antibody [EPR20545] (ab213363)

Overview

  • Product name

    Anti-CD68 antibody [EPR20545]
    See all CD68 primary antibodies
  • Description

    Rabbit monoclonal [EPR20545] to CD68
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human CD68 aa 200-300. The exact sequence is proprietary.
    Database link: P34810

  • Positive control

    • WB: Human tonsil, fetal liver and fetal spleen lysates; THP-1 and U937 whole cell lysates. IHC-P: Human tonsil, liver and cervix carcinoma. ICC/IF: THP-1 and U937 cells.
  • 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.

Properties

  • 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

    EPR20545
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab213363 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. Detects a band of approximately 110 kDa (predicted molecular weight: 37 kDa).
IHC-P 1/8000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/100.

Target

  • Function

    Could play a role in phagocytic activities of tissue macrophages, both in intracellular lysosomal metabolism and extracellular cell-cell and cell-pathogen interactions. Binds to tissue- and organ-specific lectins or selectins, allowing homing of macrophage subsets to particular sites. Rapid recirculation of CD68 from endosomes and lysosomes to the plasma membrane may allow macrophages to crawl over selectin-bearing substrates or other cells.
  • Tissue specificity

    Highly expressed by blood monocytes and tissue macrophages. Also expressed in lymphocytes, fibroblasts and endothelial cells. Expressed in many tumor cell lines which could allow them to attach to selectins on vascular endothelium, facilitating their dissemination to secondary sites.
  • Sequence similarities

    Belongs to the LAMP family.
  • Post-translational
    modifications

    N- and O-glycosylated.
  • Cellular localization

    Cell membrane and Endosome membrane. Lysosome membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • CD 68 antibody
    • CD68 antibody
    • CD68 antigen antibody
    • CD68 molecule antibody
    • CD68_HUMAN antibody
    • DKFZp686M18236 antibody
    • gp11 antibody
    • Gp110 antibody
    • LAMP4 antibody
    • Macrophage antigen CD68 (microsialin) antibody
    • MACROPHAGE ANTIGEN CD68 antibody
    • Macrosialin antibody
    • SCARD1 antibody
    • Scavenger receptor class D member 1 antibody
    see all

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) of human liver tissue.

    Panel A showed merged staining of anti-Collagen VI (ab182744), anti-CD68 (ab213363) and anti-Lamin B1 (ab229025). Anti-Collagen VI (panel B, green signal) stained on extracellular matrix, anti-CD68 (panel C, red signal) stained on Kupffer cells, and anti-Lamin B1 (panel D, magenta signal) stained on nuclear envelope.

    The section was incubated in three rounds of staining with ab182744, ab213363 and ab229025 for 30 mins at room temperature. Each round was followed by tyramide signal amplification with the appropriate fluorophore.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

    DAPI was used as a nuclear counter stain. A ready to use anti-Rabbit and Mouse Polymer HRP was used as a secondary. 

  • Immunofluorescent analysis of 100% methanol-fixed THP-1 (human monocytic leukemia cell line) cells labeling CD68 with ab213363 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on THP-1 cells.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue, labeling CD68 with ab213363 at 1/8000 dilution, followed by Goat anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on macrophages of human tonsil is observed (PMID: 19543531). 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.

  • Lanes 1-3 & 5 : Anti-CD68 antibody [EPR20545] (ab213363) at 1/1000 dilution
    Lane 4 : Anti-CD68 antibody [EPR20545] (ab213363) at 1/5000 dilution

    Lane 1 : Human fetal liver lysate at 20 µg
    Lane 2 : Human tonsil lysate at 20 µg
    Lane 3 : Human fetal spleen lysate at 20 µg
    Lane 4 : THP-1 (human monocytic leukemia cell line) whole cell lysate at 10 µg
    Lane 5 : U937 (human histiocytic lymphoma cell line) whole cell lysate at 10 µg

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

    Developed using the ECL technique.

    Predicted band size: 37 kDa
    Observed band size: 110 kDa
    why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1/2/4/5: 30 seconds; Lane 3: 3 minutes.

    The observed molecular weight is consistent with the literature (PMID:18405323; PMID:11739566; PMID: 16710801).

  • Immunofluorescent analysis of 100% methanol-fixed U937 (human histiocytic lymphoma cell line) cells labeling CD68 with ab213363 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on U937 cells.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human cervical carcinoma tissue labeling CD68 with ab213363 at 1/8000 dilution, followed by Goat anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on macrophages of human cervical carcinoma is observed (PMID: 12118106). 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.

  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CD68 with ab213363 at 1/5000 dilution. No blocking step performed. Anti-Rabbit HRP polymer was used as the secondary detection system. Heat-mediated antigen retrieval was performed using EDTA based pH 9.0 buffer.

  • Immunohistochemical analysis of paraffin-embedded human liver tissue labeling CD68 with ab213363 at 1/8000 dilution, followed by Goat anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on Kupffer cells of human liver is observed (PMID: 12118106). 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.

References

ab213363 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rhesus monkey Tissue sections (colon)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris-EDTA pH 9
Permeabilization
No
Specification
colon
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 2% · Temperature: 23°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jun 18 2019

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Microglia)
Permeabilization
Yes - 0.3% Triton X-100
Specification
Microglia
Blocking step
FBS as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Apr 11 2019

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