HA600051

eIF-6 Mouse Monoclonal Antibody [A6A3]

cat.: HA600051

Product Type:	Mouse monoclonal IgG2a, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, IF-Cell, FC
Clonality:	Monoclonal
Clone number:	A6A3

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	PBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	2ug/ul
Purification:	Protein G affinity purified.
Molecular weight:	Predicted band size: 27 kDa
Isotype:	IgG2a
Immunogen:	Recombinant protein within human eIF-6 aa 1-200.
Positive control:	HeLa cell lysate, HepG2 cell lysate, RAW264.7 cell lysate, NIH/3T3 cell lysate, mouse testis tissue lysate, rat kidney tissue lysate, rat testis tissue lysate, human colon cancer tissue, human kidney tissue, mouse kidney tissue, rat kidney tissue, HepG2, NIH/3T3.
Subcellular location:	Cytoplasm, Nucleus, nucleolus.
Recommended Dilutions: WB IHC-P IF-Cell FC	1:1,000 1:1,000 1:100 1:1,000
Uniprot #:	SwissProt: P56537 Human \| O55135 Mouse \| Q3KRD8 Rat
Alternative names:	b(2)gcn B(2)GCN homolog B4 integrin interactor Binding protein of beta-4 integrin CAB eIF-6 EIF3A EIF6 Eukaryotic translation initiation factor 3A Eukaryotic translation initiation factor 6 IF6_HUMAN Integrin beta 4 binding protein ITGB4BP OK/SW-cl.27 p27 beta 4 integrin binding protein p27(BBP) p27BBP RP4-614O4.1

Images

Fig1: Western blot analysis of eIF-6 on different lysates with Mouse anti-eIF-6 antibody (HA600051) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: HepG2 cell lysate
Lane 3: RAW264.7 cell lysate
Lane 4: NIH/3T3 cell lysate
Lane 5: Mouse testis tissue lysate
Lane 6: Rat kidney tissue lysate
Lane 7: Rat testis tissue lysate

Lysates/proteins at 30 µg/Lane.

Predicted band size: 27 kDa
Observed band size: 27 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 (HA600051) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Anti-Mouse IgG for IP Nano-secondary antibody (NBI02H) at 1/5,000 dilution was used for 1 hour at room temperature.

Fig2: Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Mouse anti-eIF-6 antibody (HA600051) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (HA600051) 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 human kidney tissue with Mouse anti-eIF-6 antibody (HA600051) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (HA600051) 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 Mouse anti-eIF-6 antibody (HA600051) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (HA600051) 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: Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Mouse anti-eIF-6 antibody (HA600051) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (HA600051) 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.

	Fig6: Immunocytochemistry analysis of HepG2 cells labeling eIF-6 with Mouse anti-eIF-6 antibody (HA600051) 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 Mouse anti-eIF-6 antibody (HA600051) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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 (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.
	Fig7: Flow cytometric analysis of HepG2 cells labeling eIF-6. Cells were fixed and permeabilized. Then stained with the primary antibody (HA600051, 1μg/mL) (red) compared with Mouse IgG1 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-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
	Fig8: Flow cytometric analysis of NIH/3T3 cells labeling eIF-6. Cells were fixed and permeabilized. Then stained with the primary antibody (HA600051, 1μg/mL) (red) compared with Mouse IgG1 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-Mouse IgG Secondary antibody (HA1125) 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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