I***a
发帖数: 13467 | 1 谢谢，
同样的受体在HL－60cells上的行为，
跟在macrophage 或是DC 上不一样，
但是又没有真的neutrophil 拿来试一下。 |
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a*******g
发帖数: 33 | 2 试了ice cold PBS，细胞也是岿然不动。trypsin有说会消化掉细胞表面抗原，大家做
macrophage会用trypsin、EDTA一起来消化脱壁吗？ |
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w******y
发帖数: 2504 | 3 Immunol Cell Biol. 2011 Nov 8. doi: 10.1038/icb.2011.89. [Epub ahead of
print]
Macrophage migration inhibitory factor deficiency protects pancreatic islets
from palmitic acid-induced apoptosis.
Saksida T, Stosic-Grujicic S, Timotijevic G, Sandler S, Stojanovic I.
SourceDepartment of Immunology, Institute for Biological Research 'Sinisa
Stankovic', University of Belgrade, Belgrade, Serbia.
Could anyone help me send a copy mentioned above to my email: purplewindxxh@
hotmail.com. Thanks so much. |
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w******y
发帖数: 2504 | 4 Could anyone send me a copy of the following paper to : purplewindxxh@
hotmail.com?
Int J Mol Med. 2000 Apr;5(4):397-403.
Quantitation of macrophage migration inhibitory factor (MIF) using the one-
step sandwich enzyme immunosorbent assay: elevated serum MIF concentrations
in patients with autoimmune diseases and identification of MIF in
erythrocytes.
Mizue Y, Nishihira J, Miyazaki T, Fujiwara S, Chida M, Nakamura K, Kikuchi K
, Mukai M.
SourceSapporo Immunodiagnostic Laboratory, Sapporo 001, Ja... 阅读全帖 |
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n***3
发帖数: 663 | 5 J Neuroimmunol. 2010 Jul 27;224(1-2):51-5. Epub 2010 May 31.
Ion channels in monocytes and microglia/brain macrophages: promising
therapeutic targets for neurological diseases.
Eder C.
Please send the paper to e*********[email protected]
Thanks so much! |
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h******t
发帖数: 56 | 6 对了，忘记说了。我用的是RPMI+10% heat inactivated FBS+PEN/STREP和50ng/ml GM-
CSF |
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h******t
发帖数: 56 | 7 多谢回答,我用的beads是invitrogen的untouched monocytes isolation beads，而且
cd14 positive cell 有96%+这么高, 所以我觉得应该没有多少T cell contamination. |
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H*Q
发帖数: 725 | 8 will send you some baozi as my appreciation.
J Pathol. 2012 May 18. doi: 10.1002/path.4053. [Epub ahead of print]
Anti-VEGF antibody therapy does not promote metastasis in genetically
engineered mouse tumor models.
Singh M, Couto SS, Forrest WF, Lima A, Cheng JH, Molina R, Long JE, Hamilton
P, McNutt A, Kasman I, Nannini MA, Reslan HB, Cao TC, Ho CC, Barck KH,
Carano RA, Foreman O, Eastham-Anderson J, Jubb AM, Ferrara N, Johnson L.
J Pathol. 2012 May 18. doi: 10.1002/path.4052. [Epub ahead of pr... 阅读全帖 |
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j******b
发帖数: 78 | 9 mouse mac, we used F4/80, rat anti-mouse; AbD Serotec. Worked very well in
mouse bone marrow. |
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D*a
发帖数: 6830 | 10 CD31 BD 和 F4/80请问有reference么？ |
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D*a
发帖数: 6830 | 11 CD31 BD 和 mac3 or F4/80 请问有reference么？ |
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C**S
发帖数: 522 | 13 BD的CD31抗体在paraffin切片上不好用，只能做冰冻切片。 |
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y****y
发帖数: 123 | 17 德国一家公司的CD31据说是唯一可以染老鼠paraffin的
但是好像二抗那步比较麻烦
明天去看看号
顺便问一下BD的CD31染冰冻用什么固定比较好?
formalin可以么 |
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D*a
发帖数: 6830 | 25 多谢楼上两位～剩下的包子留着我求bless的时候发了～ |
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l**********1
发帖数: 5204 | 26 LZ can also ask some chinese PD or Pre-PD at below lab:
//kingsley.stanford.edu/lab_members.html
if you still do not fix it.
from their Protocol link:
//kingsley.stanford.edu/Lab_Protocols.html
about Mouse that is amazing
NB: original hint was from former post here:
http://www.mitbbs.com/article_t/Biology/31684277.html
12 th floor |
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w***a
发帖数: 4361 | 27 physical activity如果没有变化的话，
得测Respiratory exchange ratio，看看代谢速率怎么样,fuel source主要是糖还是脂
肪。KO老鼠吃得少，但是体重不变，能量怎么平衡得首先搞清楚。
另外insulin sensitivity跟WT obese老鼠比怎么样？这个也可以提供重要线索。
如果KO老鼠更加insulin sensitive，那么很可能你敲除的基因影响了免疫细胞，比如
macrophage，T Cell,B Cell这些。 |
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l**********1
发帖数: 5204 | 28 Bingo
plus one Chinese American super Bull in this field:
Frank Hu
Harvard Medical School
his group one paper:
Malik VS et al.(2010)
Sugar-sweetened beverages, obesity, type 2 diabetes mellitus, and
cardiovascular disease risk.
Circulation. 121: 1356-1364.
link:
//www.ncbi.nlm.nih.gov/pubmed/20308626
his lab publications link:
//www.hsph.harvard.edu/faculty/frank-hu/publications/
and PubMed all papers link:
//www.ncbi.nlm.nih.gov/pubmed?term=Hu%20FB%5BAuthor%5D&cauthor=true&cauthor_
uid=2030862... 阅读全帖 |
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i****e
发帖数: 1527 | 29 听起来很有些道理...
T, B, DC, Macrophage...搞免疫的, 大家基本上都围绕这些细胞作东西，作出点突破
性的，原创性的东西不容易啊!
原 |
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J******s
发帖数: 7538 | 30 my question is how lipids meet albumin locally, because albumin should be
abundant in blood. |
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J******s
发帖数: 7538 | 31 so lipid binding albumin was from lymph, entered LN through afferent
lymphatics. |
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p****e
发帖数: 105 | 32 这个问题也困扰我很久，albumin也可以出现在interstitial fluid里，但是浓度可能
没有在serum里高，具体有多高对我很重要，但是没有文献有小鼠的数据。 |
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f**********r
发帖数: 95 | 34 在写paper, 描述一个gentamicin protection assay.老板写”We found that WT
macrophages harbored fewer viable bacteria“。虽然bacteria确实有单数和复数的
分别，但还是觉得这个读起来有点怪。这里应该说”fewer“还是”less“啊？ |
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B****w
发帖数: 48 | 35 太谢谢了，包子奉上。我的问题主要是怕人家不相信我得到的细胞是免疫细胞。
T
such
macrophage |
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w******y
发帖数: 4871 | 36 可不可以上flowcytometry啊，看看表达那些CDmarker或其它的marker,就应该知道是T,
B,NK,macrophage了吧 |
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o**d
发帖数: 3080 | 37 不光是T／B细胞，也要考虑到macrophage or DC。
For T/B cell, isolate T/b from spleen and stimulate them with CD3/CD28 for T
cell, IgM/LPS/CD40 etc for B cell. Look at proliferation and some cytokines
production (IL-2/IFN-g for T cells), will give your some ideas.
For antibody titer, get sera form mice and do ELISA for IgM/IgG/IgA, etc.
For mf or DC, try use TLRs ligands to stimulate them and look at cytokines
production (ELISA or qPCR).
Should be good for the first run screening. |
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d****i
发帖数: 2346 | 38 有同学做过这个类似的实验没有？最近做这个老不成功，想请教一些细节问题。
谢谢！ |
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W****C
发帖数: 1937 | 40 以前实验室有人做过， 把细菌培养物扔给培养的巨噬细胞然后收集划板数克隆。 还可
以用beads模拟， 用流式测吞噬的beads。 |
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c*****i
发帖数: 1392 | 42 不应该啊，无论是否有phagocytosis,至少细胞应该在。37C后，是否在显微镜下观察细
胞形态完好？考虑换buffer看看 |
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d****i
发帖数: 2346 | 43
buffer用的是RPMI+10%FBS。确实不应该，但是好几次都这样，实在不知道哪里错了。 |
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S**********e
发帖数: 1789 | 44 给你看看我们系刚进来的助理教授
Vanderbilt University School of Medicine
Fingleton, Barbara Mary , Ph.D. Assistant Professor of Cancer Biology
Publications
Barrett, CW, Fingleton, B, Williams, A, Ning, W, Fischer, MA, Washington, MK
, Chaturvedi, R, Wilson, KT, Hiebert, SW, Williams, CS. MTGR1 is required
for tumorigenesis in the murine AOM/DSS colitis-associated carcinoma model.
Cancer Res, 71(4), 1302-12, 2011.
Koon, HB, Fingleton, B, Lee, JY, Geyer, JT, Cesarman, E, Parise, RA, Egorin,
MJ, Dezube, BJ... 阅读全帖 |
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z*******6
发帖数: 679 | 46 我怎么记着inflammatory macrophage是表达lyve-1的，我不是专家，隔壁实验室天天
做，听说的... |
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p****e
发帖数: 105 | 47 Good, thanks.
Lymphatic endothelium-specific hyaluronan receptor LYVE-1 is expressed by
stabilin-1+, F4/80+, CD11b+ macrophages in malignant tumours and wound
healing tissue in vivo and in bone marrow cultures in vitro: implications
for the assessment of lymphangiogenesis. J Pathol. 209:67–77. |
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s****9
发帖数: 932 | 48 生物版讨论起千青、裙带、争authorship叽里哇啦的上万人看贴，上百人留言讨论。讨
论起真正的生物了，3个人留言，2个外行（不是专家），一个上来什么也不分析就要重
新买抗体。
我也不是专家，专家都在讨论Cui J和内幕，我贡献一下自己的2 cents。
你的staining 肯定有问题。Lyve1只表达在lymphatic endothelium上，不在
macrophage上表达。你如果把图贴出来，可能大家还能帮助你。没有图，我只能猜测有
2个可能：
1）你的F4/80antibody不好。F4/80 antibody有2个clone，一个是CI:A3-1，一个是BM8
。8M8 fix的条件比较tricky，不容易在section 上stain，FACS很好用。如果你用的是
8M8，建议改用CI:A3-1，或者仔细摸一下tissue fixation的时间。Lyve1只有一个
clone （ALY7 ），很亮也很好用。
2）即使你用错了F4/80的antibody，也不应该2个antibody co－localize，LN section
最容易出现non－specific b... 阅读全帖 |
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p****e
发帖数: 105 | 49 谢谢了。我给老鼠打了CpG ODN，淋巴结有平时3-4倍大，所以Macrophage 应该是在
inflammatory状态下，抗体的clone我从来没注意，回头看一下（我本行不是学生物的
）。我的F4/80和Lyve-1荧光大都在subcapsular sinus area，图明天贴上来。多谢各
位。
region. |
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p**t
发帖数: 160 | 50 F4/80 staining is non-specific.
Lyve-1 staining can be better.
Have a look at the attachment, which is from the paper of
Michael C. Carroll 2010 on Immunity. Blue signal is F4/80,
green is CD169. Blue signal means F4/80 positive medullary
macrophage. Although here is no higher magnification, the signal
is so different with yours.
You can also check Jason Cyster's Nature Immunology paper to see
how the signal of F4/80 in their hands. Carefully check your
abs for histology. The problem should be t... 阅读全帖 |
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