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Fluorochrome  Antibodies
Fluorochrome  Antibodies
  • Fluorochrome  Antibodies

Fluorochrome Antibodies

产品报价:询价

更新时间:2023/4/3 18:11:18

地:美国

牌:Fluorochrome

号:Antibodies

厂商性质: 生产型,

公司名称: 世联博研(北京)科技有限公司

产品关键词: Fluorochrome 抗体   Antibodies   Fluorochrome Antibodies   Fluorochrome  

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在PVN中注入荧光金后的杏仁核细胞(40X)。抗体为1 / 50,000。
VTA(将氟金注射到伏安中后40倍。


我们的氟金抗体是迄今为止最有效的氟金抗体。自从Fluorochrome在1998年首次推出以来,它已经取得了行之有效的记录,并已成为行业标准。

Ki-67抗体-W,GFAP抗体-W和BrdU抗体-W是最近推出的优秀抗体。这些抗体包括被特定封闭肽吸收的滤纸。我们认为在使用这些类型抗体的所有研究中均应使用封闭剂。只有使用封闭剂,研究人员才能验证并确信观察到的信号是预期的信号。我们已免费提供了阻断剂,该阻断剂使研究人员可以轻松,准确地进行验证。

使用指南和协议

氟金抗体使用指南和方案

在PVN中
注入Flouro-Gold后,沿着第三个左心室(40X)的下丘脑细胞
抗体为1 / 50,000。

抗体的储存和制备
您将在一个小瓶中收到氟金抗体。它是在兔中产生的多克隆抗体。小瓶将包含约100μl抗体溶液。应立即将其冷冻并保存在寒冷(-20摄氏度),黑暗,干燥的环境中。好的商用冰箱不具备无霜功能的冰柜部分就足够了。如果正确并连续冷冻,抗体溶液可以保存长达一年。

抗体溶液应保持冷冻直至准备使用。抗体的100 μl等分试样的稀释度为1/100。可以在BSA稀释剂(50 mM KPBS,0.4%Triton,1%BSA,1%NGS)中将抗体进一步稀释1 / 50,000至1 / 100,000倍,以产生50ml至100ml的工作效价。这意味着您可以将收到的溶液稀释500到1,000倍。如果使用Vector elite套件,则应处理300至1000个切片。制备后,抗体溶液可以在阴凉干燥的阴凉(4摄氏度)环境中最多保存7天。好的商用冰箱的冰箱部分就足够了。我们不建议在准备后7天以上使用该溶液。不要重新冷冻抗体溶液。

抗体的使用

可以使用几种不同的方法注射氟金,包括压力,离子电渗疗法和由许多研究人员开发的其他应用程序。参见Schmued和Fallon,《荧光金》:“具有许多独特特性的荧光逆行轴突示踪剂”,《大脑研究》,377(1986)147-154以及Pieribone和Aston-Jones,《荧光金在离子电渗疗法中的应用》。大脑深部神经核传入的研究,” Brain Research,475(1988)259-271。许多研究人员已经开发了自己的修改程序。抗体的使用不应依赖于使用Fluoro-Gold的方法。

注射氟金后,将浮动切片(我们从大鼠中注入30%的切片,灌以4%甲醛)与氟金抗体溶液在4摄氏度下孵育过夜。将切片洗涤,然后在生物素化GAR中孵育(Vector Labs)在室温下以1/1000放置1小时,然后再次洗涤。然后将切片在抗生物素蛋白/生物素(Vector Labs)中以1/1000孵育1小时,洗涤并转移至含0.16%Na2乙酸盐和0.06%H2O2的二氨基联苯胺(0.04%)和氯化镍(2.5%)中六分钟。然后将切片洗涤,固定,干燥,脱水并盖玻片。

我们的经验是,如果以上述方式存储和制备抗体,则使用Vector Elite试剂盒时,每小瓶抗体应可处理300至1000微米的白化病大鼠大脑(或类似大小的动物)切片。但是,您应该尝试使用浓度和程序来确定最适合您的情况。

Ki-67抗体-W使用指南和方案

一般:
Ki-67抗体-W对Ki-67蛋白起反应。Ki-67蛋白是正在进行的细胞增殖的内在标志物,存在于细胞周期的所有活跃阶段(G1,S,G2和有丝分裂),而静止细胞(G0)则不存在。这意味着它是确定给定细胞群体生长分数的极佳标记。它广泛用于诊断,研究和药物发现应用中。Ki-67抗体-W是多克隆的,在兔体内被育成人肽序列–TPKEKAQALEDLAGFKELFQT –在注射入兔前通过戊二醛与甲状腺球蛋白偶联。我们的抗体已用于大鼠和人脑组织中的免疫细胞化学。它将与吸收了特定肽的封闭滤纸一起出售。这使研究人员可以确认特定信号。

Ki-67抗体-W的表征


A. 7日龄大鼠海马中的Ki-67阳性细胞(10X)
B.共聚焦显微镜拍摄的海马中的Ki-67细胞(60X)
C.海马中的Ki-67阻滞(10X)

通过进行阻断研究确定特异性。将7天大的大鼠海马的相邻切片与单独的Ki-67抗体-W(A-10X)和(B-60X)或与Ki-67抗体-W预先孵育Ki-67肽(C -10X)。注意在A和B中标记,在C中没有标记,表明抗体结合的特异性。

储存和准备:
您将收到的小瓶包含大约200μl溶液(稀释度为1/100)。Ki-67抗体-W可以在BSA稀释剂(50mM KPBS,0.4%Triton,1%BSA,1%NGS)中进一步稀释至至少1 / 20,000。

抗体应储存在-20℃。还包括被特定肽吸收的用于阻塞的滤纸。可以将500μl稀释的抗体添加到试管中,以达到10μM封闭。在施用于组织之前,应在室温(RT)下孵育至少1小时。该抗体特异性地阻断该肽,而不阻断吸收在滤纸上的其他肽。

协议:
将新鲜冷冻的10-30微米切片在室温下固定在4%多聚甲醛(FA)中1小时,在磷酸盐缓冲液(PBS)中洗涤,然后在90%的10%柠檬酸钠中处理40分钟。使切片在柠檬酸钠溶液中冷却20分钟,在PBS中洗涤,并与0.1至0.3%的H 2 O 2在PBS中孵育10至30分钟。再次在PBS中洗涤组织,然后用抗生物素蛋白/生物素封闭试剂盒(Vector-cat#SP-2001)处理,洗涤并与BSA稀释剂在室温下孵育1小时。将Ki67抗体(1 / 20,000)在室温下过夜。用PBS洗涤后,将切片在RT / 1/1000(Vector labs-cat#BA1000)的生物素化GAR中孵育1小时,洗涤,然后在Avidin / Biotin / 1/1000(Vector labs-cat#PK6100)的孵育中1hr。在RT。洗涤后,用0.1M乙酸钠中的二氨基联苯胺(0.04%)和0.06%的H2O2溶液观察信号六分钟(也可以添加2.5%的氯化镍)。然后将切片用水洗涤(如果需要,可以冲洗),脱水并盖玻片。

Ki-67 Antibody-W还可以与用于可视化Fluorochrome,LLC销售的BRDU Antibody-W的协议配合使用。但是,可以使用4%FA灌注的组织代替新鲜的冷冻切片。

出版物:
Perez等人,《神经科学杂志》,2009年5月13日:29(19):6379-6387 Garcia-Fuster等人。

GFAP抗体-W使用指南和方案

概述:
GFAP抗体-W与胶质纤维酸性蛋白(GFAP)反应,胶质纤维酸性蛋白是III类中间细丝,是星形胶质细胞中中间细丝的主要组成部分,是成熟胶质细胞的细胞特异性标记物。它可用作神经干细胞和星形细胞的标志物,包括表达GFAP的多种类型的源自星形细胞的脑肿瘤。GFAP抗体-W是多克隆的,在兔体内被制备成戊二醛与甲状腺球蛋白偶联的肽序列NAGFKETRASERAE(人,大鼠和小鼠)(人,大鼠和小鼠),然后注入兔。我们的抗体已用于大鼠和人脑组织中的免疫细胞化学。它也将与吸收了特定肽的封闭滤纸一起出售。这使研究人员可以确认特定信号。

GFAP抗体-W的表征

GF_large

A.成年大鼠海马中的GFAP glail细胞(10X)
B.共聚焦显微镜拍摄的海马中的GFAP glail细胞(60X)
C.海马中的GFAP阻滞(10)

通过进行阻断研究确定特异性。将成年大鼠海马的相邻切片与单独的GFAP抗体-W(A-10X)和(B-60X)或与GFAP肽预孵育的GFAP抗体-W(C-10X)一起孵育。注意在神经胶质细胞中在A和B中标记,在C中没有标记,表明抗体结合的特异性。

储存和准备:
您将收到的小瓶含有大约200μl的1/100稀释液。抗体可以在BSA稀释剂(50mM KPBS,0.4%Triton,1%BSA,1%NGS)中进一步稀释至至少1 / 30,000。

抗体应储存在-20℃。还包括被特定肽吸收的用于阻塞的滤纸。可以将500μl稀释的抗体添加到试管中,以达到10 μM封闭。在施用于组织之前,应在室温(RT)下孵育至少1小时。该抗体特异性地阻断该肽,而不阻断吸收在滤纸上的其他肽。

协议:
将新鲜冷冻的10-30微米切片在室温下固定于4%多聚甲醛(FA)中1小时,在磷酸盐缓冲液(PBS)中洗涤,然后与0.1-0.3%H2O2的PBS孵育10至30分钟。再次在PBS中洗涤组织,然后用抗生物素蛋白/生物素封闭试剂盒(Vector-cat#SP-2001)处理,洗涤并与BSA稀释剂在室温下孵育1小时。将GFAP抗体-W(1 / 30,000)在室温下过夜。用PBS洗涤后,将切片在RT / 1/1000(Vector labs-cat#BA1000)的生物素化GAR中孵育1小时,洗涤,然后在Avidin / Biotin / 1/1000(Vector labs-cat#PK6100)的孵育中1hr。在RT。洗涤后,用0.1M乙酸钠中的二氨基联苯胺(0.04%)和0.06%的H2O2溶液观察信号六分钟(也可以添加2.5%的氯化镍)。

GFAP Antibody-W还可以与用于可视化Fluorochrome,LLC销售的BRDU Antibody-W的协议配合使用。但是,可以使用4%FA灌注的组织代替新鲜的冷冻切片。

BrdU抗体-W使用指南和方案

概述:
该抗体与合成的胸苷类似物BrdU反应,当将其注射入动物体内后,在细胞周期的S期将其掺入新合成的活跃增殖细胞的DNA链中。可以通过免疫细胞化学测量存活的细胞数量。作为评估一组细胞增殖状态的出色工具,它已成为药物发现研究的关键,尤其是在ADME / Tox研究期间进行癌症治疗以及确定细胞健康方面。BrdU抗体-W是兔抗BrdU的多克隆抗体,其与牛血清白蛋白(BSA)偶联。我们的抗体已用于大鼠脑组织的免疫细胞化学。它将与吸收有BrdU的滤纸一起销售以进行阻塞。这使研究人员可以确认特定信号。

BrdU抗体-W的表征

Br_large

A.先前注射过BrdU的成年大鼠的齿状回(60X)中的BrdU阳性细胞。在共焦显微镜B.BrdU块在齿状回中拍摄的照片

通过进行阻断研究确定特异性。将已预先注射BrdU的成年大鼠海马的相邻切片与单独的BrdU Antibody-W(A-60X)或与BrdU预孵育的BrdU Antibody-W(B-60X)一起孵育。注意在A中标记而在B中没有标记,表明抗体阻断的特异性。

储存和准备:
您将收到的小瓶含有大约200μl的1/100稀释液。BrdU Antibody-W可以在BSA稀释剂(50mM KPBS,0.4%Triton,1%BSA,1%NGS)中进一步稀释至至少1 / 30,000。

抗体应储存在-20℃。还包括被BrdU吸收的滤纸,用于阻塞。可以将500μl稀释的抗体添加到试管中,以达到10 μM封闭。应用于组织之前,应在室温(RT)下孵育至少1小时。该抗体特异性地被BrdU阻断,而不被吸收在滤纸上的其他肽阻断。

协议:
将新鲜冷冻的10-30微米切片在室温下固定在4%多聚甲醛(FA)中1小时,在磷酸盐缓冲液(PBS)中洗涤,然后用50%甲酰胺/ 2X SSC(300 mM氯化钠和30 mM柠檬酸钠处理) )在65C下放置2小时。将切片在2X SSC中洗涤,在2N HCl中于37℃温育30分钟,然后在RT下直接置于100mM硼酸钠(pH 8.5)中10分钟。将切片在PBS中洗涤,并与PBS中的0.1至0.3%H 2 O 2一起温育10至30分钟。再次在PBS中洗涤组织,然后用抗生物素蛋白/生物素封闭试剂盒(Vector-cat#SP-2001)处理,洗涤并与BSA稀释剂在室温下孵育1小时。将BrdU抗体-W(1 / 30,000)在室温下过夜。用PBS洗涤后,将切片在生物素化的GAR中于1/1000(Vector labs-cat#BA1000)于室温孵育1小时,洗涤,然后在抗生物素蛋白/生物素中以1/1000(Vector labs-cat#PK6100)在室温下孵育1小时。洗涤后,用0.1M乙酸钠中的二氨基联苯胺(0.04%)和0.06%的H2O2溶液观察信号六分钟(也可以添加2.5%的氯化镍)。然后将切片用水洗涤(如果需要,可以冲洗),脱水并盖玻片。

*注意:氟金,氟金抗体,氟红宝石,KI-67抗体-W,GFAP抗体-W和BrdU抗体-W仅用于实验室研究动物或用于体外测试。不适用于人类。这些药物仅应由经常从事神经解剖学研究和体外测试的人员或仅用于实验室研究的动物使用。



Our Antibody to Fluoro-Gold is by far the most effective antibody to Fluoro-Gold available. Since Fluorochrome first introduced it in 1998, it has a achieved a proven record of accomplishment and has become the standard in the industry.

The Ki-67 Antibody-W, GFAP Antibody-W, and BrdU Antibody-W are excellent antibodies that are recent introductions. Included with these antibodies is filter paper absorbed with a specific blocking peptide. We feel that a blocking agent should be utilized in all investigations using these types of antibodies. Only with a blocking agent can the researcher verify and have confidence that the observed signal is the intended signal. At no additional costs, we have included the blocking agent, which makes this verification easy and accurate for the researcher to implement.

Use Guide and Protocol

Fluoro-Gold Antibody Use Guide and Protocol

Hypothalamic cells along the 3rd left 
ventricle (40X) after Flouro-Gold injection 
in the PVN. Antibody is at 1/50,000.

Storage and Preparation of the Antibody
You will receive the Antibody to Fluoro-Gold in a small vial. It is a polyclonal antibody raised in rabbit. The vial will contain approximately 100μl of the antibody solution. It should be immediately frozen and stored in a cold (-20 degrees C), dark, dry environment. The freezer section, without the frost free feature, of a good commercial refrigerator should suffice. If properly and continuously frozen, the antibody solution can be stored up to one year.

The antibody solution should remain frozen until ready for use. The 100μl aliquot of the Antibody is at a dilution of 1/100. The Antibody can be further diluted 1/50,000 to 1/100,000 times in BSA diluent (50 mM KPBS, 0.4% Triton, 1% BSA, 1% NGS) to produce 50ml to 100ml of working titer. This means you can dilute the solution you receive by 500 to 1,000 times. This should treat 300 to 1000 sections if you use the Vector elite kit. After preparation, the antibody solution can be stored for up to seven days in a cool (4 degrees C), dark and dry environment. The refrigerator section of a good commercial refrigerator should suffice. We do not recommend that the solution be used beyond seven days after preparation. Do not refreeze the antibody solution.

Use of the Antibody

Fluoro-Gold can be injected using several different methods, including pressure, iontophoretic and other applications developed by a variety of researchers. See Schmued and Fallon, Fluoro-Gold: “A fluorescent retrograde axonal tracer with numerous unique properties,” Brain Research, 377 (1986) 147-154 as well as Pieribone and Aston-Jones, “The Iontophoretic Application of Fluoro-Gold for the study of afferents to deep brain nuclei,” Brain Research, 475 (1988) 259-271. Many researchers have developed their own modified procedures. Use of the antibody should not be dependent upon the methodology used to employ Fluoro-Gold.

After the Fluoro-Gold has been injected, floating sections (we used thirty um sections from a rat perfused with 4% formaldehyde) are incubated with the Fluoro-Gold Antibody solution overnight at 4 degrees C. Sections are washed, then incubated in Biotinylated GAR (Vector Labs) at 1/1000 for 1 hour at room temperature and washed again. Sections are then incubated in Avidin/Biotin (Vector Labs) at 1/1000 for 1 hour, washed and transferred to Diaminobenzidine (.04%) and Nickel Chloride (2.5%) in 0.1 M NaAcetate with 0.06% H2O2 for six minutes. Sections are then washed, mounted, dried, dehydrated and cover slipped.

It has been our experience that if stored and prepared in the manner set out above, each vial of the antibody should treat 300 to 1000 thirty um sections of the albino rat brain (or similar sized animals) if you use the Vector elite kit. However, you should experiment with the concentration and procedures to determine which best fits your circumstance.

Ki-67 Antibody-W Use Guide and Protocol

General:
Ki-67 Antibody-W reacts against the Ki-67 protein. The Ki-67 protein is an intrinsic marker of ongoing cell proliferation and is present during all active phases of the cell cycle (G1, S, G2 and mitosis) and is absent from resting cells (G0). This means it is an excellent marker for determining the growth fraction of a given cell population. It is used extensively in diagnostic, research and drug discovery applications. Ki-67 Antibody-W is polyclonal and was raised in rabbit to the human peptide sequence –TPKEKAQALEDLAGFKELFQT – that was coupled to thyroglobulin by glutaraldehyde before injection into the rabbit. Our antibody has been used for immunocytochemistry in rat and human brain tissue. It will be sold with filter paper absorbed with the specific peptide for blocking. This allows the researcher to confirm the specific signal.

Characterization of the Ki-67 Antibody-W


A. Ki-67 positive cells in the hippocampus (10X) of a 7 day old rat
B. Ki-67 cells in hippocampus (60X) taken on a confocal microscope
C. Ki-67 block in hippocampus (10X)

Specificity was ascertained by performing a blocking study. Adjacent sections from the hippocampus of a 7 day old rat were incubated with the Ki-67 Antibody-W alone (A-10X) and (B-60X) or with the Ki-67 Antibody-W preincubated with Ki-67 peptide (C-10X). Note labeling in A and B and the absence of labeling in C, indicating specificity of the antibody binding.

Storage and Preparation:
The vial you will receive contains approximately 200μl of solution at a dilution of 1/100. The Ki-67Antibody-W can be further diluted to at least 1/20,000 in BSA diluent (50mM KPBS, 0.4% Triton, 1% BSA, 1% NGS).

The antibody should be stored at -20C. Also included is filter paper absorbed with the specific peptide for blocking. 500μl of the diluted antibody can be added to the tube to achieve a 10μM block. Incubate for at least 1 hour at room temperature (RT) before applying to tissue. The antibody blocks specifically with this peptide and does not block with other peptides absorbed on filter paper.

Protocol:
Fresh frozen 10-30 micron sections are fixed in 4% paraformaldehyde (FA) for 1 hour at RT, washed in phosphate buffered saline (PBS), and then treated in 10% Sodium Citrate for 40 minutes at 90C. The sections are allowed to cool for 20 minutes in the sodium citrate solution, washed in PBS and incubated with 0.1 to 0.3% H2O2 in PBS for 10 to 30 minutes. The tissue is again washed in PBS, then treated with an Avidin/Biotin blocking kit (Vector-cat #SP-2001), washed and incubated with BSA diluent for 1 hour at RT. The Ki67 antibody (1/20,000) is applied to the tissue overnight at RT. After washing in PBS the sections are incubated in Biotinylated GAR at 1/1000 (Vector labs-cat#BA1000) for 1hr at RT, washed and then incubated in Avidin/Biotin at 1/1000 (Vector labs-cat#PK6100) for 1hr at RT. After washing the signal is visualized with Diaminobenzidine (0.04%) in 0.1M Sodium Acetate with 0.06% H2O2 for six minutes (Nickel Chloride at 2.5% may also be added). Sections are then water washed (counterstained if desired), dehydrated and cover slipped.

Ki-67 Antibody-W also works with the protocol used to visualize the BRDU Antibody-W sold by Fluorochrome, LLC. However, 4% FA perfused tissue may be used instead of fresh frozen sections.

Publications:
Perez et al, The Journal of Neuroscience, May 13, 2009:29(19):6379-6387 Garcia-Fuster et al, in preparation: Individual differences in adult hippocampal neurogensis following experimenter administration of cocaine.

GFAP Antibody-W Use Guide and Protocol

General:
GFAP Antibody-W reacts against the Glial Fibrillary Acidic Protein (GFAP), which is a class-III intermediate filament and the main constitute of intermediate filaments in astrocytes and serves as a cell specific marker for mature glial cells. It is useful as a marker of neural stem cells and astrocytic cells, including many types of brain tumor derived from astrocytic cells, which express GFAP. GFAP Antibody-W is polyclonal and was raised in rabbit to the peptide sequence NAGFKETRASERAE (human, rat and mouse) coupled to thyroglobulin by glutaraldehyde before injection into the rabbit. Our antibody has been used for immunocytochemistry in rat and human brain tissue. It will also be sold with filter paper absorbed with the specific peptide for blocking. This allows the researcher to confirm the specific signal.

Characterization of the GFAP Antibody-W

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A. GFAP glail cells in hippocampus (10X) of an adult rat
B. GFAP glail cells in hippocampus (60X) taken on a confocal microscope
C. GFAP block in hippocampus (10)

Specificity was ascertained by performing a blocking study. Adjacent sections from the hippocampus of an adult rat were incubated with the GFAP Antibody-W alone (A-10X) and (B-60X) or with GFAP Antibody-W preincubated with GFAP peptide (C-10X). Note labeling in A and B in glial cells and the absence of labeling in C, indicating specificity of the antibody binding.

Storage and Preparation:
The vial you will receive contains approximately 200μl at a dilution of 1/100. The antibody can be further diluted to at least 1/30,000 in BSA diluent (50mM KPBS, 0.4% Triton, 1% BSA, 1% NGS).

The antibody should be stored at -20C. Also included is filter paper absorbed with the specific peptide for blocking. 500μl of the diluted antibody can be added to the tube to achieve a 10 μM block. Incubate for at least 1 hour at room temperature (RT) before applying to tissue. The antibody blocks specifically with this peptide and does not block with other peptides absorbed on filter paper.

Protocol:
Fresh frozen 10-30 micron sections are fixed in 4% paraformaldehyde (FA) for 1 hour at RT, washed in phosphate buffered saline (PBS) and incubated with 0.1 to 0.3% H2O2 in PBS for 10 to 30 minutes. The tissue is again washed in PBS, then treated with an Avidin/Biotin blocking kit (Vector-cat #SP-2001), washed and incubated with BSA diluent for 1 hr at RT. The GFAP Antibody-W (1/30,000) is applied to the tissue overnight at RT. After washing in PBS the sections are incubated in Biotinylated GAR at 1/1000 (Vector labs-cat#BA1000) for 1hr at RT, washed and then incubated in Avidin/Biotin at 1/1000 (Vector labs-cat#PK6100) for 1hr at RT. After washing the signal is visualized with Diaminobenzidine (0.04%) in 0.1M Sodium Acetate with 0.06% H2O2 for six minutes (Nickel Chloride at 2.5% may also be added). Sections are then water washed (counterstained if desired), dehydrated and cover slipped.

GFAP Antibody-W also works with the protocol used to visualize the BRDU Antibody-W sold by Fluorochrome, LLC. However, 4% FA perfused tissue may be used instead of fresh frozen sections.

BrdU Antibody-W Use Guide and Protocol

General:
This antibody reacts against the synthetic thymidine analog BrdU, which when injected into an animal is incorporated during the S phase of the cell cycle into newly synthesized DNA strands of actively proliferating cells. The number of cells that survive can be measured by immunocytochemistry. As an outstanding tool to assess the proliferation state of a group of cells, it has become vital to drug discovery research, especially with cancer therapeutics and ascertaining the health of cells-during ADME/Tox studies. BrdU Antibody-W is a polyclonal raised in rabbit against BrdU conjugated to bovine serum albumin (BSA). Our antibody has been used for immunocytochemistry in rat brain tissue. It will be sold with filter paper absorbed with BrdU for blocking. This allows the researcher to confirm the specific signal.

Characterization of the BrdU Antibody-W

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A. BrdU positive cells in dentate gyrus (60X) of an adult rat previously injected with BrdU. Photo taken on confocal microscope
B. BrdU block in dentate gyrus

Specificity was ascertained by performing a blocking study. Adjacent sections from the hippocampus of an adult rat that had been previously injected with BrdU were incubated with the BrdU Antibody-W alone (A-60X) or with BrdU Antibody-W preincubated with BrdU (B-60X). Note labeling in A and the absence of labeling in B, indicating specificity of the antibody blocking.

Storage and Preparation:
The vial you will receive contains approximately 200μl at a dilution of 1/100. BrdU Antibody-W can be further diluted to at least 1/30,000 in BSA diluent (50mM KPBS, 0.4% Triton, 1% BSA, 1% NGS).

The antibody should be stored at -20C. Also included is filter paper absorbed with BrdU for blocking. 500μl of the diluted antibody can be added to the tube to achieve a 10 μM block. Incubate for at least 1hour at room temperature (RT) before applying to tissue. The antibody blocks specifically with BrdU and does not block with other peptides absorbed on filter paper.

Protocol:
Fresh frozen 10-30 micron sections are fixed in 4% paraformaldehyde (FA) for 1 hour at RT, washed in phosphate buffered saline (PBS), then treated with 50% Formamide/2X SSC (300 mM sodium chloride and 30 mM sodium citrate) at 65C for 2 hours. The sections are washed in 2X SSC, incubated in 2N HCL at 37C for 30 minutes, and then placed directly in 100mM Sodium Borate (pH 8.5) for 10 minutes at RT. The sections are washed in PBS and incubated with 0.1 to 0.3% H2O2 in PBS for 10 to 30 minutes. The tissue is again washed in PBS, then treated with an Avidin/Biotin blocking kit (Vector-cat #SP-2001), washed and incubated with BSA diluent for 1 hr at RT. The BrdU Antibody-W (1/30,000) is applied to the tissue overnight at RT. After washing in PBS the sections are incubated in Biotinylated GAR at 1/1000 (Vector labs-cat#BA1000) for 1hr at RT, washed and then incubated in Avidin/Biotin at 1/1000 (Vector labs-cat#PK6100) for 1hr at RT. After washing the signal is visualized with Diaminobenzidine (0.04%) in 0.1M Sodium Acetate with 0.06% H2O2 for six minutes (Nickel Chloride at 2.5% may also be added). Sections are then water washed (counterstained if desired), dehydrated and cover slipped.

Publications:
Garcia-Fuster et al, in preparation: Individual differences in adult hippocampal neurogensis following experimenter administration of cocaine.

*CAUTION: Fluoro-Gold, Antibody to Fluoro-Gold, Fluoro-Ruby, KI-67 Antibody-W, GFAP Antibody-W, and BrdU Antibody-W are for investigational use only in laboratory research animals or for tests in vitro. NOT FOR USE IN HUMANS. These drugs should be used only by persons regularly engaged in conducting neuroanatomical studies and tests in vitro or in animals used only for laboratory research.