HA721824

ATL3 Recombinant Rabbit Monoclonal Antibody [PSH02-47]

cat.: HA721824

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat, Monkey
Applications:	WB, IF-Cell, IHC-P, FC
Clonality:	Monoclonal
Clone number:	PSH02-47

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: 61 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within human ATL3 aa 1-488 / 541.
Positive control:	HeLa cell lysate, Jurkat cell lysate, A549 cell lysate, HepG2 cell lysate, COS-1 cell lysate, NIH/3T3 cell lysate, RAW264.7 cell lysate, Neuro-2a cell lysate, PC-12 cell lysate, L6 cell lysate, mouse liver tissue lysate, rat liver tissue lysate, mouse liver tissue, rat liver tissue, HeLa, NIH/3T3.
Subcellular location:	Endoplasmic reticulum membrane.
Recommended Dilutions: WB IF-Cell IHC-P FC	1:1,000 1:100 1:1,000 1:1,000
Uniprot #:	SwissProt: Q6DD88 Human \| Q91YH5 Mouse \| Q0ZHH6 Rat
Alternative names:	4633402C03Rik 5730596K20Rik AI465397 ATL3 ATLA3_HUMAN Atlastin 3 like Atlastin GTPase 3 Atlastin-3 AW228836 DKFZp564J0863 MGC28761

Images

Fig1: Western blot analysis of ATL3 on different lysates with Rabbit anti-ATL3 antibody (HA721824) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: Jurkat cell lysate
Lane 3: A549 cell lysate
Lane 4: HepG2 cell lysate
Lane 5: COS-1 cell lysate
Lane 6: NIH/3T3 cell lysate
Lane 7: RAW264.7 cell lysate
Lane 8: Neuro-2a cell lysate
Lane 9: PC-12 cell lysate
Lane 10: L6 cell lysate
Lane 11: Mouse liver tissue lysate
Lane 12: Rat liver tissue lysate

Lysates/proteins at 30 µg/Lane.

Predicted band size: 61 kDa
Observed band size: 61 kDa

Exposure time: 30 seconds;

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 (HA721824) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig2: Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-ATL3 antibody (HA721824) 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 (HA721824) 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.

	Fig3: Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-ATL3 antibody (HA721824) 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 (HA721824) 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: Immunocytochemistry analysis of HeLa cells labeling ATL3 with Rabbit anti-ATL3 antibody (HA721824) 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-ATL3 antibody (HA721824) 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.
	Fig5: Immunocytochemistry analysis of NIH/3T3 cells labeling ATL3 with Rabbit anti-ATL3 antibody (HA721824) 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-ATL3 antibody (HA721824) 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.

Fig6: Flow cytometric analysis of HeLa cells labeling ATL3.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721824, 1μg/mL) (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: Flow cytometric analysis of NIH/3T3 cells labeling ATL3.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721824, 1μg/mL) (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).

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