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LAMP2a Recombinant Rabbit Monoclonal Antibody [SA46-01]

Usd: 205

Specification

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Catalog# ET1601-24

LAMP2a Recombinant Rabbit Monoclonal Antibody [SA46-01]

Application

  • WB

  • IHC-P

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

LAMP2a Recombinant Rabbit Monoclonal Antibody [SA46-01]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human LAMP2a aa 361-410 / 410.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IP

Molecular Weight

Predicted band size: 45 kDa

Positive Control

SK-MEL-28 cell lysate, HeLa cell lysate, JAR cell lysate, U-937 cell lysate, RAW264.7 cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, mouse liver tissue lysate, rat liver tissue lysate, rat lung tissue lysate, human kidney tissue, human liver tissue, human pancreas tissue, mouse kidney tissue, mouse placenta tissue, mouse pancreas tissue, rat kidney tissue.

Conjugation

unconjugated

Clone Number

SA46-01

RRID

AB_3069607

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:5,000

  • IHC-P

  • 1:500

  • IP

  • Use at an assay dependent concentration.

Target

Function

Lysosome-associated membrane protein 2 (LAMP2), also known as CD107b (Cluster of Differentiation 107b) and Mac-3, is a human gene. Its protein, LAMP2, is one of the lysosome-associated membrane glycoproteins. The protein encoded by this gene is a member of a family of membrane glycoproteins. This glycoprotein provides selectins with carbohydrate ligands. It may play a role in tumor cell metastasis. It may also function in the protection, maintenance, and adhesion of the lysosome. Alternative splicing of the gene produces three variants - LAMP-2A, LAMP-2B and LAMP-2C. LAMP-2A is the receptor for chaperone-mediated autophagy. Recently it has been determined that antibodies against LAMP-2 account for a fraction of patients who get a serious kidney disease termed focal necrotizing glomerulonephritis. LAMP-2B is associated with Danon disease.

Background References

1. Guan, JJ. et al. 2015. DRAM1 regulates apoptosis through increasing protein levels and lysosomal localization of BAX. Cell death & disease. 6: e1624.

2. Gu, G. et al. 2013. Ubiquitin E3 Ligase A20 is Required in Degradation of Microbial Superantigens in Vascular Endothelial Cells. Cell Biochem. Biophys. 66: 649-655.

Sequence Similarity

Belongs to the LAMP family.

Tissue Specificity

Isoform LAMP-2A is highly expressed in placenta, lung and liver, less in kidney and pancreas, low in brain and skeletal muscle. Isoform LAMP-2B is detected in spleen, thymus, prostate, testis, small intestine, colon, skeletal muscle, brain, placenta, lung, kidney, ovary and pancreas and liver. Isoform LAMP-2C is detected in small intestine, colon, heart, brain, skeletal muscle, and at lower levels in kidney and placenta.

Post-translational Modification

O- and N-glycosylated; some of the 16 N-linked glycans are polylactosaminoglycans.

Subcellular Location

Cell membrane, Endosome membrane, Lysosome membrane

UNIPROT

SwissProt: P13473 Human

SwissProt: P17047 Mouse

SwissProt: P17046 Rat

Synonyms

CD107 antigen-like family member B antibody

CD107b antibody

LAMP 2 antibody

Lamp 2a antibody

LAMP-2 antibody

LAMP2 antibody

LAMP2_HUMAN antibody

Lysosome-associated membrane glycoprotein 2 antibody

Lysosome-associated membrane protein 2 antibody

Images

  • Western blot analysis of LAMP2a on different lysates with Rabbit anti-LAMP2a antibody (<a href="/products/ET1601-24" style="font-weight: bold;text-decoration: underline;">ET1601-24</a>) at 1/5,000 dilution.<br /><br />Lane 1: SK-MEL-28 cell lysate (20 µg/Lane)<br />Lane 2: HeLa cell lysate (20 µg/Lane)<br />Lane 3: JAR cell lysate (20 µg/Lane)<br />Lane 4: U-937 cell lysate (20 µg/Lane)<br />Lane 5: RAW264.7 cell lysate (15 µg/Lane)<br />Lane 6: NIH/3T3 cell lysate (15 µg/Lane)<br />Lane 7: PC-12 cell lysate (15 µg/Lane)<br />Lane 8: Mouse liver tissue lysate (30 µg/Lane)<br />Lane 9: Rat liver tissue lysate (30 µg/Lane)<br />Lane 10: Rat lung tissue lysate (30 µg/Lane)<br /><br />Predicted band size: 45 kDa<br />Observed band size: 70-140 kDa<br /><br />Exposure time: Lane 1-4: 2 minutes 37 seconds; Lane 5-10: 5 minutes;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1601-24" style="font-weight: bold;text-decoration: underline;">ET1601-24</a>) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of LAMP2a on different lysates with Rabbit anti-LAMP2a antibody (ET1601-24) at 1/5,000 dilution.

    Lane 1: SK-MEL-28 cell lysate (20 µg/Lane)
    Lane 2: HeLa cell lysate (20 µg/Lane)
    Lane 3: JAR cell lysate (20 µg/Lane)
    Lane 4: U-937 cell lysate (20 µg/Lane)
    Lane 5: RAW264.7 cell lysate (15 µg/Lane)
    Lane 6: NIH/3T3 cell lysate (15 µg/Lane)
    Lane 7: PC-12 cell lysate (15 µg/Lane)
    Lane 8: Mouse liver tissue lysate (30 µg/Lane)
    Lane 9: Rat liver tissue lysate (30 µg/Lane)
    Lane 10: Rat lung tissue lysate (30 µg/Lane)

    Predicted band size: 45 kDa
    Observed band size: 70-140 kDa

    Exposure time: Lane 1-4: 2 minutes 37 seconds; Lane 5-10: 5 minutes;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1601-24) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-LAMP2a antibody (<a href="/products/ET1601-24" style="font-weight: bold;text-decoration: underline;">ET1601-24</a>) at 1/500 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1601-24" style="font-weight: bold;text-decoration: underline;">ET1601-24</a>) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-LAMP2a antibody (ET1601-24) at 1/500 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1601-24) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue with Rabbit anti-LAMP2a antibody (<a href="/products/ET1601-24" style="font-weight: bold;text-decoration: underline;">ET1601-24</a>) at 1/500 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1601-24" style="font-weight: bold;text-decoration: underline;">ET1601-24</a>) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue with Rabbit anti-LAMP2a antibody (ET1601-24) at 1/500 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1601-24) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-LAMP2a antibody (<a href="/products/ET1601-24" style="font-weight: bold;text-decoration: underline;">ET1601-24</a>) at 1/500 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1601-24" style="font-weight: bold;text-decoration: underline;">ET1601-24</a>) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-LAMP2a antibody (ET1601-24) at 1/500 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1601-24) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • LAMP2a was immunoprecipitated from 0.2 mg SK-MEL-28 cell lysate with <a href="/products/ET1601-24" style="font-weight: bold;text-decoration: underline;">ET1601-24</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/ET1601-24" style="font-weight: bold;text-decoration: underline;">ET1601-24</a> at 1/2,000 dilution. HRP Conjugated Rabbit IgG kappa light chain antibody (<a href="/products/M1208-2" style="font-weight: bold;text-decoration: underline;">M1208-2</a>) at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: SK-MEL-28 cell lysate (Input)<br />Lane 2: <a href="/products/ET1601-24" style="font-weight: bold;text-decoration: underline;">ET1601-24</a> IP in SK-MEL-28 cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/ET1601-24" style="font-weight: bold;text-decoration: underline;">ET1601-24</a> in SK-MEL-28 cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 20 seconds; ECL: K1801

    LAMP2a was immunoprecipitated from 0.2 mg SK-MEL-28 cell lysate with ET1601-24 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1601-24 at 1/2,000 dilution. HRP Conjugated Rabbit IgG kappa light chain antibody (M1208-2) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: SK-MEL-28 cell lysate (Input)
    Lane 2: ET1601-24 IP in SK-MEL-28 cell lysate
    Lane 3: Rabbit IgG instead of ET1601-24 in SK-MEL-28 cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 20 seconds; ECL: K1801

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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