IHC image of ab16667 staining Ki67 in a section of formalin-fixed paraffin-embeddedrat spleen tissue. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0).. The section was then incubated with ab16667, 1/200 dilution and detected using Goat Anti-Rabbit IgG H&L (HRP) antibody. DAB was used as the chromogen. The section was then counterstained with haematoxylin.
Lanes 1 - 3: Merged signal (red and green). Green - ab16667 observed at 359 kDa. Red - loading control, ab130007 observed at 125 kDa.
ab16667 was shown to react with Ki67 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab255407 (knockout cell lysate ab263762) was used. Wild-type and Ki67 knockout samples were subjected to SDS-PAGE. ab16667 and Anti-Vinculin antibody [VIN-54] (ab130007) were incubated overnight at 4°C at 1 in 100 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
IHC image of ab16667 staining Ki67 in a section of formalin-fixed paraffin-embedded normal human tonsil tissue. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0).. The section was then incubated with ab16667, 1/200 dilution and detected using Goat Anti-Rabbit IgG H&L (HRP) antibody. DAB was used as the chromogen. The section was then counterstained with haematoxylin.
Fluorescence multiplex immunohistochemical analysis of normal human tonsil tissue (formalin-fixed paraffin-embedded section).
Merged staining of anti-PD1 (ab237728; orange; Opal™520), anti-PDL1 (ab237726; green; Opal™540), anti-CD68 (ab192847; yellow; Opal™570), anti-CD3 (ab16669; red; Opal™620), anti-Ki67 (ab16667; light blue; Opal™650) and anti-PanCK (ab7753; grey; Opal™690).
The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 7-color automation IHC kit (NEL821001KT, Akoya Biosciences®).
The section was incubated in six rounds of staining; in the order of ab237728 (1/500 dilution), ab237726 (1/500 dilution), ab192847 (1/300 dilution), ab16669 (1/300 dilution), ab16667 (1/200 dilution) and ab7753 (1/200 dilution); each using a separate fluorescent tyramide signal amplification system.
Sodium citrate antigen retrieval (Leica ER1, pH6.0, 30 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity.
DAPI (dark blue) was used as a nuclear counter stain.
Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra Polaris.
This image was generated from the hybridoma version.
IHC image of ab16667 staining Ki67 in a section of formalin-fixed paraffin-embedded mouse spleen tissue. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0).. The section was then incubated with ab16667, 1/200 dilution and detected using Goat Anti-Rabbit IgG H&L (HRP) antibody. DAB was used as the chromogen. The section was then counterstained with haematoxylin.
ab16667 staining Ki67 in wild-type HAP1 cells (top panel) and Ki67 knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol for 5 minutes, permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1 hour. The cells were then incubated with ab16667 at 1/250 dilution and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1 hour withGoat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibodyat 2μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
This image was generated from the hybridoma version.
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1000 dilution (2 µg/mL).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1000 dilution (2 µg/mL).
This image was generated from the hybridoma version.
Effect of newborn anoxia and PD156707 on proliferation and binucleation of neonatal cardiomyocytes
A representative image of cardiomyocytes stained with α-actinin (green), Ki-67 (red), and Hoescht (blue).
4% paraformaldehyde-fixed, Triton X-100 permeabilized rat cardiomyocytes were stained for Ki67 using ab16667 at 1/100 dilution in ICC/IF.
(From Figure 3A of Paradis et al)
This image was generated from the hybridoma version.
Immunocytochemistry/ Immunofluorescence analysis of human cardiac stem cells labelingKi67 withab16667 at 1/250 dilution. Cells were fixed in paraformaldehyde and permeabilized withTriton x-100, 0.01%. Cells were blocked in BSA for 1 hour at room temperature. A polyclonal chicken anti-rabbit Alex Fluor® 488 secondary antibody was used at 1/500 dilution.
This image was generated from the hybridoma version.
See Abreview
ab16667 staining Ki67 in human testis by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed using citrate buffer. Samples were then blocked with 1% BSA for 10 minutes at 21ºC followed by incubation with the primary antibody for2 hours at 1/100. A biotin-conjugated goat anti-rabbit polyclonal was used as secondary antibody at a 1/250 dilution.
This image was generated from the hybridoma version.
See Abreview
ab16667 staining Ki67 - Proliferation Marker in human HEp-2 cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and permeabilized with Triton X-100 0.25% in PBS. Samples were incubated with primary antibody (1/50 in DPBS) for 1 hour at 21°C. An Atto488-conjugated Donkey anti-rabbit polyclonal (1/50) was used as the secondary antibody.
This image was generated from the hybridoma version.
See Abreview
Abcam 位于英国的剑桥科学园,成立于1998年,专门生产和分销研究型抗体。在线目录 (www.abcam.cn) 已有超过 120,000 种抗体和试剂,并不断添加,供应予全球百多个国家。
为使研究员更容易找到蛋白质研究试剂产品,在2011并购了美国的 MitoSciences 公司,加强了免疫分析方面的产品供应;同年也并购了英国的 Ascent Scientific 公司,开展了生化试剂的供应。在2012年并购了美国的 Epitomics 公司,成为一家有领导地位的 RabMAbs® 供应商。
Abcam 的目标是给世界上最好的抗体建立最大的在线目录,为各地科学家提供尖端产品,成为各国科学界的重要伙伴,为所有产品提供技术支持来使客户获得预期的结果,为提供高质量的抗体来指向尽可能多的靶蛋白,尽所能在尽可能多的应用和物种中检测每种抗体。
Abcam 专注于特定的研究领域,包括肿瘤研究、心血管研究、染色质和基因调控研究、免疫学、内参对照、微生物学、神经科学、细胞核信号转导研究、RNAi 研究、干细胞研究、亚细胞标记物等。