Recombinant Anti-CD68 antibody [KP1] - BSA and Azide free (ab233172)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Mouse monoclonal [KP1] to CD68 - BSA and Azide free
- Suitable for: Mass Cytometry, ICC/IF, IHC-P, WB
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-CD68 antibody [KP1] - BSA and Azide free
See all CD68 primary antibodies -
Description
Mouse monoclonal [KP1] to CD68 - BSA and Azide free -
Host species
Mouse -
Tested applications
Suitable for: Mass Cytometry, ICC/IF, IHC-P, WBmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat, Rabbit -
Immunogen
Tissue, cells or virus. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Human spleen lysate, THP-1 whole cell lysate IHC-P: Human tonsil and liver tissue. ICC/IF: THP-1 cells. IMC: Human tonsil tissue
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General notes
ab233172 is the carrier-free version of ab955.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
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.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
KP1 -
Isotype
IgG1 -
Light chain type
kappa -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Immunohistochemistry reagents
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab233172 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 |
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Mass Cytometry | (1) |
Use at an assay dependent concentration.
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ICC/IF |
1/50.
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IHC-P |
1/3000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB |
1/1000. Detects a band of approximately 110 kDa (predicted molecular weight: 37 kDa).
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Notes |
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Mass Cytometry
Use at an assay dependent concentration. |
ICC/IF
1/50. |
IHC-P
1/3000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/1000. Detects a band of approximately 110 kDa (predicted molecular weight: 37 kDa). |
Target
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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
modificationsN- and O-glycosylated. -
Cellular localization
Cell membrane and Endosome membrane. Lysosome membrane. - Information by UniProt
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Database links
- Entrez Gene: 968 Human
- Entrez Gene: 12514 Mouse
- Entrez Gene: 287435 Rat
- Omim: 153634 Human
- SwissProt: P34810 Human
- SwissProt: P31996 Mouse
- Unigene: 647419 Human
- Unigene: 15819 Mouse
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Alternative names
- CD 68 antibody
- CD68 antibody
- CD68 antigen antibody
see all
Images
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Lanes 1 & 3 : Anti-CD68 antibody [KP1] (ab955) at 1/1000 dilution
Lanes 2 & 4 : Anti-CD68 antibody [RM1031] (ab303565) at 1/1000 dilution
Lanes 1-2 : Human spleen tissue lysate
Lanes 3-4 : THP-1 (Human monocytic leukemia monocyte) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 37 kDa
Observed band size: 110 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation (ab955).
Blocking and diluting buffer and concentration: 5% NFDM /TBST
Exposure time: Lane1: 60 seconds; Lane2: 5 seconds; Lane3: 180 seconds; Lane4: 7 seconds
ab303565 works better than ab955 in western blot testing.
We suggest optimizing experimental protocols (increasing lysate amount, using lower dilution or higher sensitivity ECL substrate) to improve results when using ab955 in western blot.
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Imaging Mass Cytometry™ (IMC™) image of human tonsil tissue stained with Anti-CD68 antibody [KP1]. ab233172 (carrier-free antibody, purified) was metal-conjugated using a Maxpar® Antibody Labeling Kit from Fluidigm. Immunostaining was performed according to Fluidigm’s protocols. Briefly, slides were subject to deparaffinization and heat-induced epitope retrieval, followed by overnight incubation at 4°C with an antibody cocktail containing metal-tagged antibodies in blocking buffer. Slides were subsequently washed with 0.2% Triton-X and 1x PBS, counterstained with Cell-ID™ Intercalator-Ir diluted at 1/400 in 1x PBS for 30 min at room temperature, rinsed for 5 min with distilled H2O, and air-dried prior to IMC™ acquisition. IMC™ acquisition was performed using the Fluidigm Hyperion™ Imaging System.
Imaging Mass Cytometry™, IMC™, Cell-ID™, Hyperion™ and Maxpar® are trademarks of Fluidigm Canada
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Immunohistochemical analysis of paraffin-embedded human liver tissue labeling CD68 with ab233172 at 1/3000 dilution, followed by Goat Anti-mouse IgG H&L (HRP) ready to use. Cytoplasmic staining on Kupffer cells of human liver (PMID: 12118106) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-mouse IgG H&L (HRP) ready to use.
Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
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Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CD68 with ab233172 at 1/3000 dilution, followed by Goat Anti-mouse IgG H&L (HRP) ready to use. Cytoplasmic staining on macrophages of human tonsil (PMID: 19543531) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-mouse IgG H&L (HRP) ready to use.
Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
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Immunofluorescent analysis of 100% methanol-fixed THP-1 (human monocytic leukemia cell line) cells labeling CD68 with ab233172 at 1/50 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining in THP-1 cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with Recombinant Anti-beta IV Tubulin antibody [EPR16775] (ab179504) at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080) secondary antibody at 1/1000 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113) secondary antibody at 1/1000 dilution.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
References (1)
ab233172 has been referenced in 1 publication.
- McDowell SAC et al. Neutrophil oxidative stress mediates obesity-associated vascular dysfunction and metastatic transmigration. Nat Cancer 2:545-562 (2021). PubMed: 35122017