HA722302

LAMP1 Recombinant Rabbit Monoclonal Antibody [PSH05-55]

cat.: HA722302

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Mouse
Applications:	WB, IF-Cell, IHC-P, FC, IP, IF-Tissue, IHC-Fr
Clonality:	Monoclonal
Clone number:	PSH05-55

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 44 kDa
Isotype:	IgG
Immunogen:	Recombinant protein.
Positive control:	C2C12 cell lysate, NIH/3T3 cell lysate, RAW264.7 cell lysate, NIH/3T3, mouse colon tissue, mouse kidney tissue, mouse brain tissue.
Subcellular location:	Lysosome membrane, Endosome membrane, Late endosome membrane, Cell membrane, Cytolytic granule membrane.
Recommended Dilutions: WB IF-Cell IHC-P FC IP IF-Tissue IHC-Fr	1:1,000 1:100 1:1,000 1:1,000 1-2μg/sample 1:200 1:200
Uniprot #:	SwissProt: P11438 Mouse
Alternative names:	CD107 antigen like family member A CD107 antigen-like family member A CD107a CD107a antigen LAMP 1 LAMP-1 LAMP1 LAMP1_HUMAN LAMPA LGP120 lgpA Lysosomal membrane glycoprotein 120KD Lysosomal Associated Membrane Protein 1 Lysosome associated membrane glycoprotein 1 Lysosome-associated membrane glycoprotein 1 Lysosome-associated membrane protein 1 OTTHUMP00000040663

Images

Fig1: Western blot analysis of LAMP1 on different lysates with Rabbit anti-LAMP1 antibody (HA722302) at 1/1,000 dilution.

Lane 1: C2C12 cell lysate
Lane 2: NIH/3T3 cell lysate
Lane 3: RAW264.7 cell lysate
Lane 4: RAW264.7 cell lysate treated with deglycosylation

Lysates/proteins at 30 µg/Lane.

Predicted band size: 44 kDa
Observed band size: 100-120 kDa

Exposure time: 9 seconds; ECL: K1801;

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 (HA722302) at 1/1,000 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.

Fig2: Immunocytochemistry analysis of NIH/3T3 cells labeling LAMP1 with Rabbit anti-LAMP1 antibody (HA722302) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-LAMP1 antibody (HA722302) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

	Fig3: Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-LAMP1 antibody (HA722302) at 1/1,000 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 (HA722302) at 1/1,000 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-LAMP1 antibody (HA722302) at 1/1,000 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 (HA722302) at 1/1,000 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.
	Fig5: Immunofluorescence analysis of frozen mouse brain tissue with Rabbit anti-LAMP1 antibody (HA722302) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA722302, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
	Fig6: Flow cytometric analysis of NIH/3T3 cells labeling LAMP1. Cells were fixed and permeabilized. Then stained with the primary antibody (HA722302, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Fig7: LAMP1 was immunoprecipitated from 0.2 mg NIH/3T3 cell lysate with HA722302 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using HA722302 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: NIH/3T3 cell lysate (input)
Lane 2: HA722302 IP in NIH/3T3 cell lysate
Lane 3: Rabbit IgG instead of HA722302 in NIH/3T3 cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 3 seconds; ECL: K1801

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.