SKU PA1518
Size 100μg/vial
Reactivity Mouse, Rat
Clonality Polyclonal
Host Rabbit
Ig Isotype N/A
Applications IF, IHC-P, IHC-F, WB

Overview

Product Name Anti-CD68 Antibody
SKU/Catalog Number PA1518
Storage & Handling At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time.Avoid repeated freezing and thawing.
Size 100μg/vial
Description Rabbit IgG polyclonal antibody for Macrosialin(CD68) detection. Tested with WB, IHC-P, IHC-F, IF in Mouse;Rat.
Cite This Product Anti-CD68 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1518)
Host Rabbit
Contents/Buffer Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Form Lyophilized
Immunogen A synthetic peptide corresponding to a sequence in the middle region of mouse CD68(312-326aa AFCITRRRQSTYQPL), different from the related rat sequence by one amino acid.
Reactivity Mouse, Rat

Assay Details

Assay Dilutions Overview

Concentration: Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Western blot, 0.1-0.5μg/ml, Rat, Mouse
Immunohistochemistry(Paraffin-embedded Section), 0.5-1μg/ml, Mouse, Rat, By Heat
Immunohistochemistry(Frozen Section), 0.5-1μg/ml, Mouse, Rat, -
Immunofluorescence, 2μg/ml, Rat

Boster's Secondary Antibodies And IHC, WB Kits

The following reagents are used to generate the images below.

Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(P) and IHC(F).

Images And Assay Conditions

Figure 2. IHC analysis of CD68 using anti- CD68 antibody (PA1518).
CD68 was detected in paraffin-embedded section of rat liver tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti- CD68 Antibody (PA1518) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

Figure 3. IHC analysis of CD68 using anti- CD68 antibody (PA1518).
CD68 was detected in paraffin-embedded section of Rat Spleen tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti- CD68 Antibody (PA1518) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

Figure 4. IHC analysis of CD68 using anti- CD68 antibody (PA1518).
CD68 was detected in paraffin-embedded section of Mouse Liver tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti- CD68 Antibody (PA1518) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

Figure 1. Western blot analysis of CD68 using anti- CD68 antibody (PA1518).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: Rat Spleen Tissue Lysate
Lane 2: Mouse Spleen Tissue Lysate
Lane 3: Mouse RAW246.7 Tissue Lysate
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti- CD68 antigen affinity purified polyclonal antibody (Catalog # PA1518) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system.

Figure 5. IF analysis of CD68 using anti- CD68 antibody (PA1518)
CD68 was detected in paraffin-embedded section of rat liver tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution ) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/mL rabbit anti- CD68 Antibody (PA1518) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Figure 6. IF analysis of CD68 using anti- CD68 antibody (PA1518)
CD68 was detected in paraffin-embedded section of rat liver tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution ) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/mL rabbit anti- CD68 Antibody (PA1518) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Figure 7. IF analysis of CD68 using anti- CD68 antibody (PA1518) CD68 was detected in paraffin-embedded section of mouse liver tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution ) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/mL rabbit anti- CD68 Antibody (PA1518) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Target Info

Protein Target Info (Source: Uniprot.org)

Uniprot Id P31996
Gene Name Cd68
Protein Name Macrosialin
Tissue Specificity Expressed in tissue macrophages and to a lesser extent in dendritic cells.
Alternative Names Macrosialin;CD68;Cd68;
Subcellular Localization Isoform Long: Endosome membrane; Single-pass type I membrane protein. Lysosome membrane; Single-pass type I membrane protein.
Molecular Weight 34818 MW

*if product is indicated to react with multiple species, protein info is based on the human gene.

Ontology

Protein 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.
Research Areas Mouse, Rat

*You can search these to find other products in these research areas.
Background CD68, cluster of differentiation, is a 110-kD transmembrane glycoprotein that is highly expressed by human monocytes and tissue macrophages. CD68 is a member of a family of hematopoietic mucin-like molecules that includes leukosialin/CD43 and stem cell antigen CD34. The CD68 gene is mapped to 17p13.1. Immunohistochemistry can be used to identify the presence of CD68, which is found in the cytoplasmic granules of a range of different blood cells. It is particularly useful as a marker for the various cells of the macrophage lineage, including monocytes, histiocytes, giant cells, Kupffer cells, and osteoclasts. This allows it to be used to distinguish diseases of otherwise similar appearance, such as the monocyte/macrophage and lymphoid forms of leukaemia(the latter being CD68 negative). Its presence in macrophages also makes it useful in diagnosing conditions related to proliferation or abnormality of these cells, such as malignant histiocytosis, histiocytic lymphoma, and Gaucher's disease.

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Polyclonal antibody for SR D1/CD68 detection. Host: Rabbit.Size: 100μg/vial. Tested applications: IHC-P. Reactive species: Mouse. SR D1/CD68 information: Molecular Weight: 34818 MW; Subcellular Localization: Isoform Long: Endosome membrane; Single-pass type I membrane protein. Lysosome membrane; Single-pass type I membrane protein; Tissue Specificity: Expressed in tissue macrophages and to a lesser extent in dendritic cells.
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In stock
Order Product
PA1518
Buy one primary antibody get one 0.5ml HRP or Biotin secondary antibody for free.
*Sample sizes are prepared on demand and will take extra lead time. (cannot be conjugated)
$280.00

Troubleshooting

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Publications

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Leftin A, Zhao H, Turkekul M, de Stanchina E, Manova K, Koutcher JA. Sci Rep. 2017 Sep 14;7(1):11632. doi: 10.1038/s41598-017-11899-2. Iron deposition is associated with differential macrophage infiltration and therapeutic response to iron chelati...
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Lin L, Li R, Cai M, Huang J, Huang W, Guo Y, Yang L, Yang G, Lan T, Zhu K. Oxid Med Cell Longev. 2018 Mar 18;2018:7808656. doi: 10.1155/2018/7808656. eCollection 2018. Andrographolide Ameliorates Liver Fibrosis in Mice: Involvement of TLR4/NF-κB a...
Huang L, Zhang K, Guo Y, Huang F, Yang K, Chen L, Huang K, Zhang F, Long Q, Yang Q. Sci Rep. 2017 Sep 20;7(1):11989. doi: 10.1038/s41598-017-12095-y. Honokiol protects against doxorubicin cardiotoxicity via improving mitochondrial function in mous...
Leftin A, Ben-Chetrit N, Klemm F, Joyce JA, Koutcher JA. PLoS One. 2017 Sep 12;12(9):e0184765. doi: 10.1371/journal.pone.0184765. eCollection 2017. Iron imaging reveals tumor and metastasis macrophage hemosiderin deposits in breast cancer
Pang Y, Zhang Y, Huang L, Xu L, Wang K, Wang D, Guan L, Zhang Y, Yu F, Chen Z, Xie X. Int J Mol Sci. 2017 Dec 19;18(12). pii: E2766. doi: 10.3390/ijms18122766. Effects and Mechanisms of Total Flavonoids from Blumea balsamifera (L.) DC. on Skin Wou...
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Liu Z,, Xu S,, Huang X, Wang J, Gao S, Li H, Zhou C, Ye J, Chen S, Jin Zg, Liu P. Br J Pharmacol. 2015 Dec;172(23):5661-75. Doi: 10.1111/Bph.13068. Epub 2015 Feb 27. Cryptotanshinone, An Orally Bioactive Herbal Compound From Danshen, Attenuates At...
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Customer Q&As

  • Q: Do you offer BSA-free antibodies? Keyword: Bovine serum albumin, carrier protein, conjugation
    A: Yes, please contact us at support@bosterbio.com for more information about BSA-free antibodies and availability. The new BSA-free formula uses trehalose as a replacement to BSA. We have tested many alternative chemicals and found that trehalose protects the antibodies the best.
  • Q: Is your western blot protocol provided from the website applicable for all your antibodies? Keyword: applications, WB
    A: The protocol is applicable for all our antibodies in WB, the NC Membrane(0.45μm or 0.22μm) and transfer time(70 mins or 50 mins) depends on the protein molecular weight, details can be found in included protocol.
  • Q: Can I conjugate markers to this antibody? Can I link custom conjugates to this antibody? Keyword: conjugation
    A: The antibody is stored with BSA and cannot be conjugated with markers. Carrier free antibodies are available upon request. Please contact support@bosterbio.com
  • Q: Has this antibody been tested for cross-reactivity with human samples in IHC or Western Blot?
    A: We tested human samples and got negative results in IHC and WB.
  • Q: What should I use for negative control?
    A: Please contact us for negative control suggestions. You can also check expression databases such as genecards, uniprot etc. Due to logistic reasons, we do not sell serum or lysates that we use internally for positive or negative control.
  • Q: Where can I find troubleshooting information? What should I do if I have unexpected bands, high background, no signal, weak signal
    A: You can find Boster's troubleshoot guides under tech support tab. Please contact us for further assistance on troubleshooting your experiment.
  • Q: What is the immunogen sequence of this antibody? Is this antibody polyclonal or monoclonal?
    A: You can find the immunogen sequence under "
  • Q: What is the expected band size? Why is it different than the observed band size?
    A: The expected band size is predicted on the size of the protein. The actual band size may be affected by a few other factors including but not limited to:<br>1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected<br>2. Post-translational cleavage: this can cause smaller bands and or multiple bands <br><br>3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody. <br><br>4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.<br>5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher., <br>
  • Q: What is the suggested dilution ratio for Western Blot (WB), Immunohistochemistry (IHC) and or ELISA standards? What is the optimal pH for the sample?
    A: Check the datasheet for the product for details on dilution ratios for different experiments. You can find the datasheet button on the right side of the product page.
  • Q: What is the protocol you used for your Western blotting (WB) and Immunohistochemistry (IHC)?
    A: Check our protocols under the tech support tab.
  • Q: What are some alternative names that could be used to describe this product?
    A: Some common names include but are not limited to cd68 antibody, cd 68 antibody
  • Q: What are some alternative names that could be used to describe this product?
    A: Some common names include but are not limited to cd68 antibody, cd 68 antibody
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