Overview

  • Product name
    Anti-CD68 antibody [FA-11]
    See all CD68 primary antibodies
  • Description
    Rat monoclonal [FA-11] to CD68
  • Host species
    Rat
  • Specificity
    ab53444 detects surface CD68 at low levels in resident mouse peritoneal macrophages which can be enhanced with thioglycollate stimulation.
  • Tested applications
    Suitable for: ICC/IF, WB, IP, IHC-Fr, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse
  • Immunogen

    Tissue, cells or virus corresponding to Mouse CD68.

  • Positive control
    • IHC-Fr: Mouse lung, spleen and heart tissue sections; ICC/IF: RAW 246.7 cell line. Flow Cyt: Mouse peritoneal macrophages.
  • General notes

    Although some customers have had success with this antibody in IHC-P, we are unable to obtain positive results in this application and so cannot recommend it for IHC-P. We batch test the antibody in IHC-Fr.

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

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.
  • Storage buffer
    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 6.97% L-Arginine
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Purification notes
    Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant.
  • Clonality
    Monoclonal
  • Clone number
    FA-11
  • Isotype
    IgG2a
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab53444 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 Use a concentration of 5 µg/ml.
WB Use at an assay dependent concentration. Predicted molecular weight: 36 kDa.
IP Use at an assay dependent concentration.
IHC-Fr Use a concentration of 0.1 - 5 µg/ml.
Flow Cyt 1/50 - 1/100.

Use 10µl of the suggested working dilution to label 106 cells in 100µl. Membrane permeabilisation is required for this application.

ab18450 - Rat monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

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

  • Dual fluorescence combining IL-6 with markers for macrophages (CD68).

    O, P, and Q, dual labeling of IL-6 (red) and marker of macrophage (green) in db/db mouse heart tissues. Arrows in Q show the specific CD68 staining with absence of IL-6 staining. (R and S) Negative control: arrows show the absence of staining in vessels with control IgG and without primary antibodies. (T) Staining of nuclei with DAPI (blue) in heart tissues of the db/db mice.

    To identify and localize IL-6 protein in coronary arterioles, transverse sections of the mouse heart were stained using markers of endothelial cells, vascular smooth muscle cells, and macrophages. Freshly isolated hearts were embedded and frozen in OCT and sectioned at 5 μm. Slides were incubated with blocking solution (10% donkey serum in PBS) and permeabilized (0.1% Triton X-100 in PBS). Primary antibodies to IL-6 (goat polyclonal 15 micro g/ml, AF-406-NA; R&D) or macrophage marker CD68 (rat monoclonal, 1:1000, ab53444; Abcam) were used for sequential double immunofluorescence staining. Secondary antibodies were conjugated with the fluorophores FITC or Texas red. Sections were mounted in an anti-fading agent (Slowfade gold with DAPI; Invitrogen), and then the slides were observed and analyzed with a fluorescence microscope (IX81; Olympus) with a x40 objective (0.90 numerical aperture). For negative controls, primary antibodies were replaced with goat polyclonal IgG (Abcam), rabbit polyclonal IgG (GeneTex), and rat monoclonal IgG (Abcam) isotype controls at the same concentration. The specificity of the primary antibody was confirmed as the absence of immunofluorescence staining signals in the IL-6-/- mice.

    Diabetic mice (db/db).

     

  • Formaldehyde-fixed, frozen mouse lung tissue sections stained for CD68 using ab53444 at a 1/250 dilution in immunohistochemical analysis. Tissue sections were blocked using 1% BSA as a blocking agent for 10 minutes at 21°C. Primary antibody was incubated for 2 hours at 21°C. Secondary antibody was a biotin-conjugated goat anti-rat IgG at 1/250 dilution.

    See Abreview

  • IHC image of CD68 staining in mouse lung frozen tissue section. The section was incubated with ab53444, 0.1µg/ml, overnight at 4C. A goat anti-Rat biotinylated secondary antibody was used to detect the primary, and visualized using an ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

  • ab53444 staining CD68 in mouse spleen tissue by Immunohistochemistry (Frozen sections). Antibody was detected with HRP-conjugated Goat anti-Rat IgG, showing staining in the red pulp.
  • ab53444 stained RAW246.7 cells. The cells were 4% formaldehyde fixed for 10 minutes at room temperature and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab53444 at 5µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Rat IgG H&L (Alexa Fluor® 488) preadsorbed (ab150165) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

  • Staining of permeabilised mouse peritoneal macrophages with Rat anti mouse CD68 (ab53444).
  • ab53444 staining mouse heart tissue by Immunohistochemistry (IHC-frozen sections). Tissue underwent fixation in paraformaldehyde, no permeabilization and blocked in 1% serum for 10 minutes at 20°C. The primary antibody used undiluted and incubated with sample for 16 hours at 20°C. A Biotin conjugated goat polyclonal to rat Ig, diluted 1/100 was used as the secondary.  

    See Abreview

  • ab53444 at 1/100 dilution staining CD68 in mouse spleen tissue by immunohistochemistry (frozen sections). Sections were acetone fixed prior to blocking in 8% milk for 40 minutes at 36°C and then incubated with ab53444 for 20 hours at 4°C. A biotin conjugated goat polyclonal to rat Ig, diluted 1/400, was used as the secondary antibody.

    See Abreview

  • ab53444 staining CD68 in murine heart tissue by Immunohistochemistry (Frozen sections).
    Tissue was fixed in acetone, permeabilized using 0.3% Triton, blocked with 10% serum for 30 minutes at 20°C, then incubated with ab53444 at a 1/1000 dilution for 16 hours at 4°C. The secondary used was a FITC conjugated goat anti-rat polyclonal, used at a 1/1000 dilution.

    See Abreview

References

This product has been referenced in:
  • Medelin M  et al. Bridging pro-inflammatory signals, synaptic transmission and protection in spinal explants in vitro. Mol Brain 11:3 (2018). Read more (PubMed: 29334986) »
  • Saranchova I  et al. Type 2 Innate Lymphocytes Actuate Immunity Against Tumours and Limit Cancer Metastasis. Sci Rep 8:2924 (2018). Read more (PubMed: 29440650) »
See all 93 Publications for this product

Customer reviews and Q&As

1-10 of 24 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Acomys (spiny mouse) Tissue sections (Spleen)
Specification
Spleen
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Acetone

Miss. Crystal Jones

Verified customer

Submitted Nov 17 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate pH6.0
Permeabilization
Yes - TritonX-100
Specification
lung
Blocking step
Invitrogen DAB kit as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
Fixative
Formaldehyde

Miss. Yuna Son

Verified customer

Submitted Mar 10 2016

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step
Serum as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 25°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer
Sample
Rat Tissue sections (Spinal Cord section)
Specification
Spinal Cord section
Permeabilization
Yes - 0.2% Triton X-100
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Mar 25 2015

Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing
Sample
Mouse Cell lysate - whole cell (Raw264.7)
Specification
Raw264.7
Treatment
100 ng/ml LPS
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Aug 19 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Antigen retrieval step
Heat mediated
Sample
Mouse Tissue sections (Aorta)
Specification
Aorta
Permeabilization
No
Fixative
10% Formalin

Abcam user community

Verified customer

Submitted Jul 25 2014

Application
Immunohistochemistry (Frozen sections)
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Sample
Mouse Tissue sections (Lung)
Specification
Lung
Permeabilization
No
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Oct 04 2013

Answer

Thank you for contacting Abcam.

As we discussed on the phone, the immunogen sequences for ab53444, ab9089, and ab3283 are unknown due to all 3 immunogens arising from tissue and cell preparations.

For CD47 (ab9089 and ab3283) the human protein atlas information for detected presence in cells/tissue is displayed below
CD47 in tissue: http://www.proteinatlas.org/ENSG00000196776/normal
CD47 in cell lines: http://www.proteinatlas.org/ENSG00000196776/cell

For ab53444, the THP1 cell line that I mentioned on the phone would not be suitable as a positive control since it is a human cell line and ab53444 is raised against mouse CD68.

Potential positive controls for each protein are listed below:
ab53444: mouse peritoneal macrophages, mouse spleen tissue,

ab9089 and ab3283: human tonsil, A431 cell lysate (ab7909, https://www.abcam.com/a431-human-epithelial-carcinoma-cell-line-whole-cell-lysate-ab7909.html), Jurkat cell lysate (ab7899, https://www.abcam.com/jurkat-human-t-cell-lymphoblast-like-cell-line-whole-cell-lysate-ab7899.html),

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

Read More
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH6 (Ventana mRcc)
Permeabilization
No
Specification
Lung
Fixative
10% Neutral Buffered Formalin

Abcam user community

Verified customer

Submitted Oct 18 2012

Answer

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one vial of ab125212.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More

Question
Answer

Thank you for contacting us.

Your credit note ID is XXXXXX.

I am sorry that this antibody did not perform as stated on the datasheet, I have asked our Finance department to issue a credit note for you. The credit note may be used in one of the following ways:

(1) Redeemed against the original invoice if this hasn't already been paid.
(2) Held on the account for use against a future order.
(3) A full refund can be offered where no other invoices are outstanding.

Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge.

To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website.

The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice

Read More

1-10 of 24 Abreviews or Q&A

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