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Product Info Summary
| SKU: | PA1518 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Mouse, Rat |
| Host: | Rabbit |
| Application: | Flow Cytometry, IF, IHC, WB |
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Product info
Product Name
Anti-Macrosialin CD68 Antibody Picoband®
View all CD68/SR-D1 Antibodies
SKU/Catalog Number
PA1518
Size
100 μg/vial
Form
Lyophilized
Description
Superstar antibody: Boster’s anti mouse CD68 antibody (PA1518) is among its top 10 bestselling antibodies, and one of the most cited mouse/rat CD68 antibodies on the market. Because of this antibody’s high demand, we have subsequently developed a rabbit monoclonal anti CD68 antibody (M00602-1) targetting a similar epitope.
High specificity: PA1518 reacts with murine macrosialin/CD68, and has been validated with Western blotting and confirmed its stellar specificity. CD68 is a highly glycosylated protein, and its expected western blot molecular weight is between 80kDa to 110kDa, depending on glycosylation level. PA1518’s observed MW in WB is near 90-100kDa. Testing on negative control tissues showed no significant bands (images available on request) or staining. See more info in the positive and negative control design section. Based on immunogen sequence homology, this antibody is not expected to cross react with other proteins from the LAMP family, which complies with QC testing observations.
Great for CD68 IHC: This antibody produces clean and specific immunobiological stains in both mouse and rat tissues. Our QC team has validated it on spleen and liver tissues of both mice and rats. Click product images for more details on experiment conditions.
More about CD68: CD68 is a cell surface marker often used to identify macrophages and other cell types in the monocyte lineage. It is a transmembrane protein that binds to electins and selectins and plays a role in macrophage homing movement. Check out the CD68 biomarker page for more information on CD68 and view all CD68 antibodies, ELISA kits and proteins.
The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.Storage & Handling
Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-Macrosialin CD68 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1518)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
A synthetic peptide corresponding to a sequence in the middle region of mouse CD68, different from the related rat sequence by one amino acid.
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
PA1518 is reactive to CD68 in Mouse, Rat
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Observed Molecular Weight
90-100 kDa
Calculated molecular weight
34818 MW
Background of CD68/SR-D1
CD68 is a heavily glycosylated glycoprotein that is highly expressed in macrophages and other mononuclear phagocytes. Macrosialin, if not glycosylated, has the main sequence that consists of 354 amino acids with an expected molecular weight of 37.4 kD. CD68, which can be located in the cytoplasmic granules of a variety of blood cells and myocytes, can be detected via immunohistochemistry. It's particularly effective as a marker for monocytes, histiocytes, giant cells, Kupffer cells, and osteoclasts, all of which belong to the macrophage lineage. CD68 is expressed in tissue macrophages and to a lesser extent in dendritic cells. CD68 is involved in tissue macrophage phagocytic activities, as well as intracellular lysosomal metabolism and extracellular cell-cell and cell-pathogen interactions. It works by binding to tissue and organ-specific lectins or selectins, enabling macrophage subsets to be directed to specific locations. Macrophages may be able to crawl over selectin-bearing substrates or other cells if CD68 is rapidly recirculated from endosomes and lysosomes to the plasma membrane.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
PA1518 is guaranteed for Flow Cytometry, IF, IHC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.1-0.5μg/ml, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Mouse, Rat, By Heat
Immunofluorescence, 5μg/ml, Mouse
Flow Cytometry(Fixed), 1-3 μg/1x106 cells, Mouse
Positive Control
WB: rat spleen tissue, mouse spleen tissue, mouse RAW2647 whole cell
IHC: mouse liver tissue, mouse spleen tissue, rat spleen tissue, rat lung tissue, rat lung tissue, rat liver tissue
IF: mouse spleen tissue
FCM: RAW2647 cell
Validation Images & Assay Conditions
Click image to see more details
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 30 ug of sample under reducing conditions.
Lane 1: rat spleen tissue lysates,
Lane 2: mouse spleen tissue lysates,
Lane 3: mouse RAW264.7 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for CD68 at approximately 90-100 kDa. The expected band size for CD68 is at 37 kDa.
Click image to see more details
Figure 2. IHC analysis of CD68 using anti-CD68 antibody (PA1518).
CD68 was detected in a paraffin-embedded section of mouse liver tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-CD68 Antibody (PA1518) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 3. IHC analysis of CD68 using anti-CD68 antibody (PA1518).
CD68 was detected in a paraffin-embedded section of mouse spleen tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-CD68 Antibody (PA1518) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 4. IHC analysis of CD68 using anti-CD68 antibody (PA1518).
CD68 was detected in a paraffin-embedded section of rat spleen tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-CD68 Antibody (PA1518) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 5. IHC analysis of CD68 using anti-CD68 antibody (PA1518).
CD68 was detected in a paraffin-embedded section of rat lung tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-CD68 Antibody (PA1518) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 6. IHC analysis of CD68 using anti-CD68 antibody (PA1518).
CD68 was detected in a paraffin-embedded section of rat lung tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-CD68 Antibody (PA1518) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 7. IHC analysis of CD68 using anti-CD68 antibody (PA1518).
CD68 was detected in a paraffin-embedded section of rat liver tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-CD68 Antibody (PA1518) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 8. IF analysis of CD68 using anti-CD68 antibody (PA1518).
CD68 was detected in a paraffin-embedded section of mouse spleen tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 5 μg/mL rabbit anti-CD68 Antibody (PA1518) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127)) was used as secondary antibody at 1:500 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.
Click image to see more details
Figure 9. Flow Cytometry analysis of RAW264.7 cells using anti-CD68 antibody (PA1518).
Overlay histogram showing RAW264.7 cells stained with PA1518 (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-CD68 Antibody (PA1518, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Protein Target Info & Infographic
Gene/Protein Information For CD68 (Source: Uniprot.org, NCBI)
Gene Name
CD68
Full Name
Macrosialin
Weight
34818 MW
Superfamily
LAMP family
Alternative Names
Macrosialin;CD68;Cd68; CD68 GP110, LAMP4, SCARD1 CD68 molecule macrosialin|CD68 |macrophage CD68|scavenger receptor class D, member 1
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on CD68, check out the CD68 Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for CD68: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-Macrosialin CD68 Antibody Picoband® (PA1518)
Hello CJ!
PA1518 has been cited in 56 publications:
*The publications in this section are manually curated by our staff scientists. They may differ from Bioz's machine gathered results. Both are accurate. If you find a publication citing this product but is missing from this list, please let us know we will issue you a thank-you coupon.
HPMA–Copolymer Nanocarrier Targets Tumor-Associated Macrophages in Primary and Metastatic Breast Cancer
The immunofluorescence detection of the CD68 was performed on tumor specimens using the Discovery XT processor to confirm the co-staining of macrophages with the HPMA copolymer P-Alexa647-FA. Using IF and confocal microscopy the P-Alexa647-FA signal within TAMs costained with CD68 from both primary and metastatic tumor tissue specimens.
Su W, Han HH, Wang Y, Zhang B, Zhou B, Cheng Y, Rumandla A, Gurrapu S, Chakraborty G, Su J, Yang G, Liang X, Wang G, Rosen N, Scher HI, Ouerfelli O, Giancotti FG. The Polycomb Repressor Complex 1 Drives Double-Negative Prostate Cancer Metastasis by Coordinating Stemness and Immune Suppression. Cancer Cell. 2019 Aug 12;36(2):139-155.e10. doi: 10.1016/j.ccell.2019.06.009. Epub 2019 Jul 18. PMID: 31327655; PMCID: PMC7210785.
Species: Mouse
A somatic mutation in erythro-myeloid progenitors causes neurodegenerative disease
Immunostaining for CD68, IBA1, YFP and phosphorylated (p)ERK demonstrated that microglial clusters represented the accumulation of EMP-derived (YFP+) pERK+ microglia. The pathophysiological consequences of a somatic BRAF(V600E) mutation in different haematopoietic lineages are markedly distinct. BRAF(V600E) expression in HSCs can cause a tumoral disease, but results in a neurodegenerative inflammatory disorder when expressed in EMP, thereby linking the pathophysiology of clonal and neurodegenerative disorders.
Species: Mouse
Myeloid-derived growth factor inhibits inflammation and alleviates endothelial injury and atherosclerosis in mice
Authors: Meng B,Li Y,Ding Y,Xu X,Wang L,Guo B,Zhu B,Zhang J,Xiang L,Dong J,Liu M,Xiang L,Xiang G.
Species: Mouse
PA1518 usage in article: APP:IHC, SAMPLE:AORTIC TISSUE, DILUTION:1:200
Protective effects of Longhu Rendan on chronic liver injury and fibrosis in mice
Authors: Guizhi Yang,Shengwen Li,Jiahua Jin,Yuanyuan Xuan,Liqin Ding,Minxia Huang,Jun Liu,Biye Wang, Tian Lan
This study aimed at assessing the effects of Longhu Rendan (LHRD), a Chinese traditional patent medicine, on liver fibrosis and its potential mechanism. Immunohistochemical staining of CD68 showed positive staining to investigate the effect of LHRD on hepatic inflammation in mice. CD68, which was used as the marker of Kupffer cells, was downregulated after the treatment of LHRD. Through the activation of Kupffer cells, TNF-α and IL-1β are produced and then amplified chronic liver inflammation leading to liver fibrosis and cirrhosis. This suggests that LHRD is a potential medicine for the treatment of liver fibrosis.
Species: Mouse
PA1518 usage in article: APP:WB, SAMPLE:LIVER TISSUE, DILUTION:1:1000
Combined topical and systemic administration with human adipose-derived mesenchymal stem cells (hADSC) and hADSC-derived exosomes markedly promoted cutaneous wound healing and regeneration.
Author: Zhou Y,Zhao B,Zhang XL,Lu YJ,Lu ST,Cheng J,Fu Y,Lin L,Zhang NY,Li PX,Zhang J,Zhang J.
Species: Human,Mouse
PA1518 usage in article: APP:IHC, SAMPLE:SKIN TISSUE, DILUTION:1:200
Tauroursodeoxycholic Acid Alleviates Secondary Injury in Spinal Cord Injury Mice Through Reducing Oxidative Stress
Authors:Yonghui Hou,Jiyao Luan,Tiancheng Deng et al.
Species: Mouse
PA1518 usage in article: APP:IF, SAMPLE:CORTICAL NEURONS, DILUTION:1:300
Schistosoma japonicum-derived peptide SJMHE1 promotes peripheral nerve repair through a macrophage-dependent mechanism
Authors:Ma Y,Wei C,Qi X,Pu Y,Dong L,Xu L,Zhou S,Zhu J,Chen X,Wang X,Su C
This study investigated whether helminths or helminth-derived molecules might have the potential to improve peripheral nerve repair. It was demonstrated that schistosome-derived SJMHE1 promoted peripheral myelin growth and functional regeneration via a macrophage-dependent mechanism and simultaneously increased the induction of M2 macrophages. The CD68 antibody from Bosterbio was used as the macrophage marker for this study. The findings indicated that schistosome-derived SJMHE1 enhances damaged peripheral nerve repair in a macrophage-dependent manner and concomitantly increases M2 macrophage polarization, highlighting the therapeutic potential of SJMHE1 for promoting peripheral nerve repair.
Species: Mouse,Rat
PA1518 usage in article: APP:IF, SAMPLE:NERVE CONDUITS, DILUTION:1:200
Shi D,Wu J,Wu Y,Lin X,Xu C,Lian X. High-Fat Diet-Related Obesity Promotes Urethane-Induced Lung Tumorigenesis in C57BL/6J Mice. Front Oncol.2021 Feb 23;11:620993.doi:10.3389/fonc.2021. 620993.PMID:33708630;PMCID:PMC7942226.
Species: Human,Mouse
PA1518 usage in article: APP:IHC, SAMPLE:TRACHEA, DILUTION:1:100
High-Fat Diet-Related Obesity Promotes Urethane-Induced Lung Tumorigenesis in C57BL/6J Mice.
Authors: Shi D,Wu J,Wu Y,Lin X,Xu C,Lian X.
The study aimed to investigate whether high-fat diet (HFD)-induced obesity influences the susceptibility to chemical-induced lung tumorigenesis, a HFD feeding condition was combined with a multi-dose urethane-induced lung tumorigenesis model using C57BL/6J mice. Lung tissue slices were incubated with antibodies against CD68 from Bosterbio for the IHC assay. The findings showed that the role of HFD-induced obesity on lung tumorigenesis was elucidated both in vivo and in vitro in this study, and highly expressed leptin mediated the activation of STAT3 pathway was likely involved in the mechanism.
Species: Human,Mouse
PA1518 usage in article: APP:IHC, SAMPLE:TRACHEA, DILUTION:1:100
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Customer Q&As
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10 Customer Q&As for Anti-Macrosialin CD68 Antibody Picoband®
Question
What Are CD68 Macrophages?
Verified customer
Asked: 2021-10-20
Answer
CD68 is a protein highly expressed by cells in the monocyte lineage by circulating macrophages, and by tissue macrophages such as Kupffer cells, microglia.
Boster Scientific Support
Answered: 2021-10-20
Question
Our team were content with the WB result of your anti-CD68 antibody. However we have seen positive staining in leukocyte isoform short: cell membrane using this antibody. Is that expected? Could you tell me where is CD68 supposed to be expressed?
Verified Customer
Verified customer
Asked: 2020-02-10
Answer
According to literature, leukocyte does express CD68. Generally CD68 expresses in isoform short: cell membrane. Regarding which tissues have CD68 expression, here are a few articles citing expression in various tissues:
Adipose tissue, Pubmed ID: 16625196
Liver, Pubmed ID: 19159218
Platelet, Pubmed ID: 16263699
Skin, Pubmed ID: 15489334
Spleen, and Umbilical cord blood, Pubmed ID: 14702039
Boster Scientific Support
Answered: 2020-02-10
Question
What tissue types is PA1518 known to stain for?
Verified customer
Asked: 2019-12-04
Answer
The Anti-Macrosialin CD68 Antibody (PA1518) stains the following tissue types: Macrophage Dendritic cells Brain Spleen Mammary gland
Boster Scientific Support
Answered: 2019-12-04
Question
We have seen staining in mouse adipose tissue. What should we do? Is anti-CD68 antibody supposed to stain adipose tissue positively?
Verified Customer
Verified customer
Asked: 2019-12-03
Answer
From what I have seen in literature adipose tissue does express CD68. From what I have seen in Uniprot.org, CD68 is expressed in leukocyte, spleen umbilical cord blood, adipose tissue, skin, platelet, liver, among other tissues. Regarding which tissues have CD68 expression, here are a few articles citing expression in various tissues:
Adipose tissue, Pubmed ID: 16625196
Liver, Pubmed ID: 19159218
Platelet, Pubmed ID: 16263699
Skin, Pubmed ID: 15489334
Spleen, and Umbilical cord blood, Pubmed ID: 14702039
Boster Scientific Support
Answered: 2019-12-03
Question
Which amino acid range is the immunogen sequence of PA1518 from?
Verified customer
Asked: 2019-11-29
Answer
For the Anti-Macrosialin CD68 Antibody (PA1518), 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.
Boster Scientific Support
Answered: 2019-12-02
Question
Has this antibody been tested for cross-reactivity with human samples in IHC or Western Blot?
Verified Customer
Verified customer
Asked: 2019-11-29
Answer
We tested human samples and got negative results in IHC and WB.
Boster Scientific Support
Answered: 2019-11-29
Question
Do you have an IHC application on Ventana machine for PA1518?
Verified customer
Asked: 2019-07-03
Answer
Our lab hasn't used the Ventana machine for our antibodies. We suggest to watch some on-line vedio or read some related articles. http://www.path.queensu.ca/qlmp/sites/webpublish.queensu.ca.qlmpwww/files/files/Ventana%20Pamphlet.pdf
Boster Scientific Support
Answered: 2019-07-05
Question
Our lab used your anti-CD68 antibody for WB on spleen umbilical cord blood in the past. I am using mouse, and I plan to use the antibody for IF next. My lab would like examining spleen umbilical cord blood as well as adipose tissue in our next experiment. Do you have any suggestion on which antibody would work the best for IF?
Verified Customer
Verified customer
Asked: 2018-05-18
Answer
I looked at the website and datasheets of our anti-CD68 antibody and it seems that PA1518 has been validated on mouse in both WB and IF. Thus PA1518 should work for your application. Our Boster satisfaction guarantee will cover this product for IF in mouse even if the specific tissue type has not been validated. We do have a comprehensive range of products for IF detection and you can check out our website bosterbio.com to find out more information about them.
Boster Scientific Support
Answered: 2018-05-18
Question
Our lab want to know about using your anti-CD68 antibody for autocrine signaling studies. Has this antibody been tested with western blotting on rat spleen tissue? We would like to see some validation images before ordering.
R. Evans
Verified customer
Asked: 2016-09-23
Answer
We appreciate your inquiry. This PA1518 anti-CD68 antibody is tested on rat spleen tissue, tissue lysate, mouse spleen tissue. It is guaranteed to work for IF, IHC-P, IHC-F, WB in mouse, rat. Our Boster guarantee will cover your intended experiment even if the sample type has not been be directly tested.
Boster Scientific Support
Answered: 2016-09-23
Question
We are currently using anti-CD68 antibody PA1518 for rat tissue, and we are satisfied with the IHC-F results. The species of reactivity given in the datasheet says mouse, rat. Is it possible that the antibody can work on zebrafish tissues as well?
K. Carter
Verified customer
Asked: 2016-01-08
Answer
The anti-CD68 antibody (PA1518) has not been validated for cross reactivity specifically with zebrafish tissues, but there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in zebrafish you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2016-01-08
