重组Anti-CD74抗体[CLIP/3127R] (ab270265)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [CLIP/3127R] to CD74
- Suitable for: IHC-P, WB, Flow Cyt
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
-
产品名称
Anti-CD74抗体[CLIP/3127R]
参阅全部 CD74 一抗 -
描述
兔单克隆抗体[CLIP/3127R] to CD74 -
宿主
Rabbit -
经测试应用
适用于: IHC-P, WB, Flow Cytmore details -
种属反应性
与反应: Human
预测可用于: Mouse, Baboon不与反应: Rat -
免疫原
Tissue, cells or virus corresponding to Human CD74. SU-DHL-4 lymphoma cells.
-
阳性对照
- IHC-P: Human tonsil tissue. Flow: Raji cells. WB: Raji and HEK-293T cell lysate.
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. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.2
Preservative: 0.05% Sodium azide
Constituents: PBS, 0.05% BSA -
Concentration information loading...
-
纯度
Protein A/G purified -
纯化说明
Purified from Bioreactor Concentrate by Protein A/G. -
克隆
单克隆 -
克隆编号
CLIP/3127R -
同种型
IgG -
研究领域
相关产品
-
Alternative Versions
-
Compatible Secondaries
-
KO cell lines
-
KO cell lysates
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab270265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 |
---|---|---|
IHC-P | (1) |
Use a concentration of 1 - 2 µg/ml. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
WB |
Use a concentration of 1 - 2 µg/ml. Predicted molecular weight: 34 kDa.
|
|
Flow Cyt |
Use 1-2µg for 106 cells.
|
说明 |
---|
IHC-P
Use a concentration of 1 - 2 µg/ml. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
Use a concentration of 1 - 2 µg/ml. Predicted molecular weight: 34 kDa. |
Flow Cyt
Use 1-2µg for 106 cells. |
靶标
-
功能
Plays a critical role in MHC class II antigen processing by stabilizing peptide-free class II alpha/beta heterodimers in a complex soon after their synthesis and directing transport of the complex from the endoplasmic reticulum to the endosomal/lysosomal system where the antigen processing and binding of antigenic peptides to MHC class II takes place. Serves as cell surface receptor for the cytokine MIF. -
序列相似性
Contains 1 thyroglobulin type-1 domain. -
细胞定位
Cell membrane. Endoplasmic reticulum membrane. Golgi apparatus > trans-Golgi network. Endosome. Lysosome. Transits through a number of intracellular compartments in the endocytic pathway. It can either undergo proteolysis or reach the cell membrane. - Information by UniProt
-
数据库链接
- Entrez Gene: 101013212 Baboon
- Entrez Gene: 972 Human
- Entrez Gene: 16149 Mouse
- Omim: 142790 Human
- SwissProt: P04233 Human
- SwissProt: P04441 Mouse
- Unigene: 436568 Human
-
别名
- CD 74 antibody
- CD74 antibody
- CD74 antigen (invariant polypeptide of major histocompatibility complex, class II antigen-associated) antibody
see all
图片
-
Formalin-fixed, paraffin-embedded human tonsil stained for CD74 using ab270265 at 2 μg/ml in imunnohistochemical analysis.
-
Flow cytometry overlay histogram showing wild-type Raji (green line) and CD74 knockout Raji cells (ab273378) stained with ab270265 (red line). The cells were incubated in 1x PBS containing 10μg/ml human IgG and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab270265) (1x106 in 100μl at 1 μg/ml) for 30 min at 4°C.
The secondary antibody Goat anti-rabbit IgG H&L (Alexa Fluor® 488, pre-adsorbed) (ab150081) was used at 1/2000 for 30 min at 4°C.
Isotype control antibody was Rabbit IgG (monoclonal) (ab172730) used at the same concentration and conditions as the primary antibody (wild-type Raji cells - black line; CD74 knockout Raji cells ab273378 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
-
All lanes : Anti-CD74 antibody [CLIP/3127R] (ab270265) at 1 µg/ml
Lane 1 : Wild-type Raji cell lysate
Lane 2 : CD74 CRISPR/Cas9 edited Raji cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : HepG2 cell lysate
Lysates/proteins at 30 µg per lane.
Performed under reducing conditions.
Predicted band size: 34 kDa
Observed band size: 35 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab270265 observed at 35 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab270265 was shown to react with CD74 in western blot. The band observed in CD74 CRISPR/Cas9 edited cell line ab273378 (CRISPR/Cas9 edited lysate ab275529) below 35 kDa is likely to represent a truncated form. This has not been investigated further. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab270265 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at 1 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 h at room temperature before imaging.
-
Flow cytometric analysis of Raji cells labeling CD74 using ab270265 at 2 μg/106 cells (Blue), followed by goat anti-mouse IgG-CF488; isotype control (Red).
-
Anti-CD74 antibody [CLIP/3127R] (ab270265) at 2 µg/ml + HEK-293T cell lysate
Predicted band size: 34 kDa
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
-
Datasheet download
文献 (0)
ab270265 尚未被引用在任何文献中。