Anti-LAMP2A抗体- Lysosome Marker
参阅全部 LAMP2A 一抗
兔多克隆抗体to LAMP2A - Lysosome Marker
Rabbit
Replenishment batches of our polyclonal antibody, ab18528 are tested in WB. Previous batches were additionally validated in ICC/IF and IHC-P. These applications are still expected to work and are covered by our Abpromise guarantee. You may also be interested in our alternative recombinant antibody, ab125068.
适用于: WB, ICC/IF, IHC-Pmore details
与反应: Mouse, Human
预测可用于: Dog, Monkey, Chinese hamster
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
WB: Human liver tissue lysate. ICC/IF: CaCo2 cells, HeLa, MCF7 and HepG2 cells. IHC-P: Human breast adenocarcinoma tissue.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Liquid
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
Immunogen affinity purified
多克隆
IgG
Abpromise™承诺保证使用ab18528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 |
|---|---|---|
| WB | (8) | Use a concentration of 0.5 - 1 µg/ml. Detects a band of approximately 105 kDa (predicted molecular weight: 100 kDa). Abcam recommends using 3% milk as the blocking agent. |
| ICC/IF | (4) | Use a concentration of 0.04 - 1 µg/ml. |
| IHC-P | (1) | Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Entrez Gene: 3920 Human
Entrez Gene: 16784 Mouse
Omim: 309060 Human
SwissProt: P13473 Human
SwissProt: P17047 Mouse
Unigene: 496684 Human
Unigene: 414016 Mouse
Unigene: 486 Mouse
Alternative splicing produces 3 isoforms.
Lamp 2a antibody
LAMP-2 antibody
LAMP2 antibody
LAMP2_HUMAN antibody
Lysosome-associated membrane glycoprotein 2 antibody
Lysosome-associated membrane protein 2 antibody
CD107 antigen-like family member B antibody
CD107b antibody
LAMP 2 antibody
Immunocytochemistry/ Immunofluorescence - Anti-LAMP2A antibody - Lysosome Marker (ab18528)
Side-by-side comparison of ICC performance using the rabbit polyclonal ab18528 and RabMab® ab125068. Staining was performed on wild-type HeLa cells (top panel) and LAMP2 knockout HeLa cells (bottom panel, available as ab255402). The cells were fixed with 4% PFA (10 min), permeabilized with 0.1% Tween-20 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab18528 or ab125068 overnight at +4°C at 3 different concentrations: 1.0 µg/mL, 0.2 µg/mL and 0.04 µg/mL. Secondary antibody incubation was at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) (shown in green) at 1/1000 and nuclear DNA was labelled with DAPI (shown in blue).
Images were acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown. Some cytoplasmic cross-reactivity is seen using ab18528 at 1.0 µg/mL, but further titration of the antibody improves the ICC staining result. The RabMab® ab125068 shows negligible non-specific staining across the dilution range. Quantification of the antibody signal was performed using a minimum of 135 cells and data are presented as mean ± SD.
Optimal dilutions/concentrations may vary across different cell types/experiment conditions and should be determined by the end user.
Immunocytochemistry/ Immunofluorescence - Anti-LAMP2A antibody - Lysosome Marker (ab18528)
Side-by-side comparison of ICC performance using the rabbit polyclonal ab18528 and RabMab® ab125068. Staining was performed on wild-type HeLa cells (top panel) and LAMP2 knockout HeLa cells (bottom panel, available as ab255402). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Tween-20 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab18528 or ab125068 overnight at +4°C at 3 different concentrations: 1.0 µg/mL, 0.2 µg/mL and 0.04 µg/mL. Secondary antibody incubation was at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) (shown in green) at 1/1000 and nuclear DNA was labelled with DAPI (shown in blue).
Images were acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown. Some cytoplasmic cross-reactivity is seen using ab18528 at 1.0 µg/mL, but further titration of the antibody improves the ICC staining result. The RabMab® ab125068 shows negligible non-specific staining across the dilution range. Quantification of the antibody signal was performed using a minimum of 180 cells and data are presented as mean ± SD.
Optimal dilutions/concentrations may vary across different cell types/experiment conditions and should be determined by the end user.
Immunocytochemistry/ Immunofluorescence - Anti-LAMP2A antibody - Lysosome Marker (ab18528)
ab18528 staining LAMP2 in MCF7 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab18528 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
Immunocytochemistry/ Immunofluorescence - Anti-LAMP2A antibody - Lysosome Marker (ab18528)
ab18528 staining Lamp2A in CaCO2 cells treated with SB 202190 (ab120638), by ICC/IF. Increase of Lamp2A expression correlates with increased concentration of SB 202190, as described in literature.
The cells were incubated at 37°C for 3 hours in media containing different concentrations of ab120638 (SB 202190) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab18528 (5 μg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2A antibody - Lysosome Marker (ab18528)
IHC image of Lamp2A staining in human breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab18528, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Western blot - Anti-LAMP2A antibody - Lysosome Marker (ab18528)
All lanes : Anti-LAMP2A antibody - Lysosome Marker (ab18528) at 1 µg/ml (blocked with 3% Milk)
Lane 1 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 2 : Lung (Mouse) Tissue Lysate
Lane 3 : Human liver tissue lysate - total protein (ab29889)
Lane 4 : Human liver left lobe tissue lysate - membrane extract (ab29086)
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 100 kDa
Observed band size: 105 kDawhy is the actual band size different from the predicted?
Additional bands at: 30 kDa, 35 kDa, 55 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 8 minutes
Abcam recommends using 3% milk as the blocking agent.
Lanes 3 and 4 are human liver tissue lysates, total protein.
