生物化学技术

Chemokines are proinflammatory cytokines that attract and activate specific types of leukocyte1. There are two main chemokine families, based on the position of the first two cysteine residues: the CC and the CXC chemokines1. Chemokines mediate their effects through interactions wi ...

Chemokines as regulators of T cell differentiation （PDF格式，81K） SanjivA.Luther & JasonG.Cyster Chemokines and disease （PDF格式，1284K）CraigGerard & BarrettJ.Rollins Viral exploitation and subversion of the immune system through chemokine mimicry (PDF格式，194K）PhilipM ...

(一)　原理　　目前用于抗体标记的荧光素主要有异硫氰酸荧光素(Fluorescein isothiocynate,FITC)或罗达明(Lissamine rhodamine B200, RB200)。在硷性条件下FITC的碳酰胺键可与抗体赖氨酸的ε氨基共价结合，标记后的抗体仍保持与相应抗原结合的能力。在荧光灯源紫外线或兰紫光激发下产生黄绿色荧光，通过在荧光显微镜下观察或流式细胞仪分析可对相应抗原进行定性、定位或定量的检测。　(二)　操作步骤　　　　将纯化 ...

1. 8 mg 辣根过氧化物酶放入1 ml玻璃瓶，加入1 ml双蒸水溶解，液体呈棕色。2. 放入一个磁力搅拌子，电磁搅拌同时逐滴缓缓加入新配制的 NaIO4 0.2 ml，室温下继续搅拌40 min，液体呈现草绿色。3. 将全部溶液用滴管装入反复用去离子水冲洗的透析袋中，4 oC 对1 mM pH 4.4的NaAc透析过夜，中间换液3－4次，每次300 ml，溶液最终呈浅棕色。4. 按照酶：抗体＝1：1的比例准备抗体，抗体浓度不小于2 mg/ml。5. 对0.01 M Na2CO3透析过夜，中间换液3－4次。6. 将 ...

Very useful for rapid and simple generation of antibodies for western blots, ELISA assays, and immunoprecipitation.--------------------------------------------------------------------------------Rabbit ImmunizationInitial Preparation Your antigen should at least have a molecular weight of 20 kDa (kil ...

Hancock Laboratory Methods,Department of Microbiology and Immunology,University of British Columbia, British Columbia, Canadahttp://www.cmdr.ubc.ca/bobh/showmethod.php?methodid=24Kill immunized mouse by CO2 asphyxia and wet throughly with ethanol. Asepti ...

The following method is recommended for purification of monoclonal antibodies produced by tissue culture (hollow fibre cartridges or roller cultures) for laboratory use. General notes: 1) Endotoxin - you must assume that all standard laboratory glassware will be coated with loads of en ...

This note is not intended as a full protocol for making monoclonal antibodies, but is to give a few hints and tips for the use of Y3/Ag1.2.3. Generally speaking the methodology is similar to making murine (eg. with NS0) monoclonal antibodies, but there are a few critical differences which are commented on b ...

We use Affigel-10, converting its functional group first to amino and then to iodoacetyl. You can also buy amino resin (it's more expensive) and start at step 6. All washes are performed on a glass-fritted filter funnel sucking until a wet cake is formed. Do not dry the resin completely or you will introduce lo ...

Materials Tumor cells that have been treated with 8-azaguanine for 48 hours are removed from the drug and grown to a maximum concentration of 500,000 cells per ml.Rats or mice that were previously immunized and then boosted IV 72 hours prior to hybridization.Media: Auto-Pow (powdered MEM from Flow ...

Protein A, derived from the cell wall of Staphylococcus aureus, and protein G, derived from the cell wall of certain strains of b-hemolytic Streptococcci, binds the Fc region of various immunoglobulines. Protein A is a 42 kDa polypeptide that binds the residues in the CH2 and CH3 domains of immunoglo ...

Borrowed from David Bowtell (bowtell@ariel.ucs.unimelb.edu.au)Transfer the srongest positive clones to a 24 well plate by scraping the adhering cells with a truncated yellow tip. Bring the volume to 1 ml using the IMDM-m medium. When cells reach confluency, freeze them ( about E6 cells per full w ...

Fusion is an important procedure to produce monoclonal antibodies (MoAb).PROTOCOLprepare the culture of myeloma cells and maintain the concentration of the cells at the level of less than 10^6 cells/ml (IMDM + 10% FCS/FBS). Just before the fusion wash 3 times in IMDM (serum free!), 1200 rpm, 5min, 37ѓC. ...

The Following Protocol can be used to couple either FITC or Biotin to IgG. The same method is used for both, and when discrepancies occur, they will be noted. 1. Dissolve 2mg of IgG in 1mL of PBS (pH 8.0) in clean 16x125mm polypropylene test tube. If needed, adjust pH to 8.0 using 0.5M carbonate buffer.2. Prepare fresh, 1 ...

Polyclonal AntibodyImmunization of Mice:Injections should be made at intervals of at least two weeks.. We have been using either of the two following adjuvants with success: Freund adjuvant and MPL+TDM adjuvant ( RIBI ImmunoChem Research, Inc. ), the latter being less hazardous than the firs ...

Materials:1) TS Buffer: 10 mM Tris pH 8.0150 mM NaCl2) TST Buffer: TS Buffer with 0.05% Tween-203) Alkaline Phosphatase Buffer: 50 mM NaCarbonate1 mM MgCl2(Bring to pH 9.8 with 1 N HCl)4) AP Substrate: p-nitrophenyl phosphate (5 mg/tablet). SigmaCat.# N93895) AP-conjugated secondary Ab: AP-Rabbit ant ...

Refer to the Western Blot Protocol for preparation of bacterially expressed protein. From a 10 ml bacterial culture, 2-4 plates can be coated for a direct ELISA. Direct ELISA:Lyse and sonicate the bacteria that express the protein of interest. If the recombinant protein is not soluble in H2O, disso ...

IntroductionImmunohistochemistry is used to identify the locationand distribution of target antigens in cells ortissues by staining with a specific antibody. Theantibody is conjugated to either a fluorescent orcolorimetric label, and the location of the labelseen through a m ...

In these techniques an antibody is used to link a cellular antigen specifically to a stain that can be more readily seen with a microscope. Detection of antigens in tissues is known as immunohistochemistry, while detection in cultured cells is generally termed immunocytochemistry. For both, ...

National Institute of Neurological Disorders and Stroke and Béla Hunyady National Institute of Mental Health，National Institutes of HealthBethesda, Maryland 20892http://intramural.nimh.nih.gov/lcmr/snge/Protocols/IHH/immuno.htmlBoth immunohistoche ...