重组Anti-Acid phosphatase抗体[EPR21787] (ab235448)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR21787] to Acid phosphatase
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
-
产品名称
Anti-Acid phosphatase抗体[EPR21787]
参阅全部 Acid phosphatase 一抗 -
描述
兔单克隆抗体[EPR21787] to Acid phosphatase -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
-
阳性对照
- WB: HepG2, SH-SY5Y, HCT 116, Jurkat, HeLa, C6 and NIH/3T3 whole cell lysates; human placenta and colon lysates. IP: HeLa whole cell lysate. IHC-P: Human colon cancer tissue; mouse colon tissue; rat colon tissue. ICC/IF: HeLa and HepG2 cells. Flow Cyt (intra): HeLa cells.
-
常规说明
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
-
形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
-
纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR21787 -
同种型
IgG -
研究领域
相关产品
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
KO cell lines
-
KO cell lysates
-
Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab235448于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
---|---|---|
Flow Cyt (Intra) |
1/50.
|
|
WB |
1/1000. Detects a band of approximately 18 kDa (predicted molecular weight: 18 kDa).
|
|
IHC-P |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
|
ICC/IF |
1/100.
|
|
IP |
1/30.
|
说明 |
---|
Flow Cyt (Intra)
1/50. |
WB
1/1000. Detects a band of approximately 18 kDa (predicted molecular weight: 18 kDa). |
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/100. |
IP
1/30. |
靶标
-
相关性
Acid phosphatases (AP) dephosphorylate phosphate groups from phosphate esters under acid conditions. Different acid phosphatase isozymes are found in different organs, and their serum levels are used as a diagnostic for disease in the corresponding organs. Elevated prostatic acid phosphatase levels may indicate the presence of prostate cancer and elevated tartrate-resistant acid phosphatase levels may indicate bone disease. -
细胞定位
ACP1: Cytoplasm. ACP2: Lysosome membrane; Single-pass membrane protein. ACP5: Lysosome. ACPP: Isoform 1: Secreted. Isoform 2: Lysosome membrane; Single-pass type I membrane protein. -
数据库链接
- Entrez Gene: 52 Human
- Entrez Gene: 53 Human
- Entrez Gene: 54 Human
- Entrez Gene: 55 Human
- Omim: 171500 Human
- Omim: 171640 Human
- Omim: 171650 Human
- Omim: 171790 Human
see all -
别名
- Acid phosphatase 1 soluble antibody
- Acid phosphatase 2 lysosomal antibody
- Acid phosphatase 5 tartrate resistant antibody
see all
图片
-
All lanes : Anti-Acid phosphatase antibody [EPR21787] (ab235448) at 1/1000 dilution
Lane 1 : Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 2 : ACP1 knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 3 : K562 (Human chronic myelogenous leukemia lymphoblast cell line ) whole cell lysate
Lane 4 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 18 kDaLanes 1 - 4: Merged signal (red and green). Green - ab235448 observed at 18 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab235448 was shown to specifically react with ACP1 (Acid phosphatase 1) in wild-type HEK 293 cells as signal was lost in ACP1 knockout cells. Wild-type and ACP1 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab235448 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
-
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labeling Acid phosphatase with ab235448 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining in human colon cancer (PMID: 25811796) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
-
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Acid phosphatase with ab235448 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and nuclear staining in HeLa cell line (PMID 26159288). The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
-
All lanes : Anti-Acid phosphatase antibody [EPR21787] (ab235448) at 1/1000 dilution
Lane 1 : HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg
Lane 2 : SH-SY5Y (human neuroblastoma cell line from bone marrow) whole cell lysate at 20 µg
Lane 3 : HCT 116 (human colorectal carcinoma cell line) whole cell lysate at 20 µg
Lane 4 : Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
Lane 5 : Human placenta lysate at 20 µg
Lane 6 : Human colon lysate at 20 µg
Lane 7 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 10 µg
Lane 8 : C6 (rat glioma cell line) whole cell lysate at 10 µg
Lane 9 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate at 10 µg
Secondary
Lanes 1-6 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Lanes 7-9 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 18 kDa
Observed band size: 18 kDaBlocking and dilution buffer: 5% NFDM/TBST.
Exposure times.
Lane 1: 37 seconds, Lanes 2-9: 3 minutes.
The molecular mass observed is consistent with what has been described in the literature (PMID:25811796).
-
Intracellular flow cytometric analysis of4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cell line labeling Acid phosphatase with ab235448 at 1/50 dilution (red) compared with a Isotype control details (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077), at 1/2000 dilution was used as the secondary antibody.
-
Acid phosphatase was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab235448 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab235448 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
Lane 1: HeLa whole cell lysate lysate 10 μg (Input).
Lane 2: ab235448 IP in HeLa whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab235448 in HeLa whole cell lysate.Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes. -
Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling Acid phosphatase with ab235448 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining in rat colon (PMID: 25811796) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
-
Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling Acid phosphatase with ab235448 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining in mouse colon (PMID: 25811796) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
-
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling Acid phosphatase with ab235448 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and nuclear staining in HepG2 cell line (PMID 26159288). The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
-
SDS download
-
Datasheet download
Certificate of Compliance
文献 (0)
ab235448 尚未被引用在任何文献中。