Antibodies, Proteins, Kits and Reagents for Life Science

257 980 ₸ Product size

100 µl 257 980 ₸ 10 µl 103 862 ₸

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Anti-LAMP2 antibody [EPR19512] (ab199946)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR19512] to LAMP2
Suitable for: WB, IHC-P
Knockout validated
Reacts with: Human

Product name

Anti-LAMP2 antibody [EPR19512]
See all LAMP2 primary antibodies
Description

Rabbit monoclonal [EPR19512] to LAMP2
Host species

Rabbit
Tested Applications & Species

Application Species
IHC-P
Human

WB
Human

See all applications and species data
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human fetal liver, fetal heart, fetal spleen, fetal kidney and placenta lysates; HeLa, HepG2, HEK-293, THP-1, Jurkat and JAR whole cell lysates. IHC-P: Human kidney and thyroid cancer tissues.
General notes
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

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, as well as customer reviews and Q&As.

Application	Species
IHC-P	Human
WB	Human

Form

Liquid
Storage instructions

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.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR19512
Isotype

IgG
Research areas

Western blot - Anti-LAMP2 antibody [EPR19512] (ab199946)

All lanes : Anti-LAMP2 antibody [EPR19512] (ab199946) at 1 µg/ml

Lane 1 : Wild-type HEK-293 cell lysate
Lane 2 : LAMP2 knockout HEK-293 cell lysate
Lane 3 : Wild-type HeLa cell lysate
Lane 4 : LAMP2 knockout HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 45 kDa

Lanes 1 - 4: Merged signal (red and green). Green - ab199946 observed at 110 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab199946 was shown to react with LAMP2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab255402 (knockout cell lysate ab263861) was used. Wild-type and LAMP2 knockout samples were subjected to SDS-PAGE. ab199946 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 µg/ml 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.
Western blot - Anti-LAMP2 antibody [EPR19512] (ab199946)

All lanes : Anti-LAMP2 antibody [EPR19512] (ab199946) at 1/2000 dilution

Lane 1 : Wild-type HEK-293 whole cell lysate
Lane 2 : LAMP2 knockout HEK-293 whole cell lysate
Lane 3 : HepG2 whole cell lysate
Lane 4 : THP1 whole cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 45 kDa

Lanes 1 - 4: Merged signal (red and green). Green - ab199946 observed at 45 kDa. Red - loading control, ab130007, observed at 130 kDa.

ab199946 was shown to recognize LAMP2 in wild-type HEK-293 cells as signal was lost at the expected MW in LAMP2 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and LAMP2 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab199946 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/2000 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2 antibody [EPR19512] (ab199946)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling LAMP2 with ab199946 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on Human kidney is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-LAMP2 antibody [EPR19512] (ab199946)

All lanes : Anti-LAMP2 antibody [EPR19512] (ab199946) at 1/2000 dilution

Lane 1 : Human fetal liver lysate
Lane 2 : Human fetal heart lysate
Lane 3 : Human fetal kidney lysate
Lane 4 : Human fetal spleen lysate
Lane 5 : Human placenta lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/100000 dilution

Predicted band size: 45 kDa
Observed band size: 110-130 kDa
why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1 and 2: 15 seconds; Lane 3,4 and 5: 5 seconds.

The molecular weight observed is consistent with what has been described in the literature (PMID: 19828315) and lots of other products.
Western blot - Anti-LAMP2 antibody [EPR19512] (ab199946)

All lanes : Anti-LAMP2 antibody [EPR19512] (ab199946) at 1/2000 dilution

Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : THP-1 (Human monocytic leukemia cell line) whole cell lysate
Lane 3 : JAR (Human placenta choriocarcinoma cell line) whole cell lysate
Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 45 kDa
Observed band size: 110-130 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1,2 and 3: 1 second; Lane 4: 5 seconds.

The molecular weight observed is consistent with what has been described in the literature (PMID: 19828315) and lots of other products.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2 antibody [EPR19512] (ab199946)

Immunohistochemical analysis of paraffin-embedded Human thyroid cancer tissue labeling LAMP2 with ab199946 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on Human thyroid cancer is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Anti-LAMP2 antibody [EPR19512] (ab199946)

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