HA722289

PABP Recombinant Rabbit Monoclonal Antibody [PSH05-51]

cat.: HA722289

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Monkey
Applications:	WB, IF-Cell, IHC-P, FC, IF-Tissue
Clonality:	Monoclonal
Clone number:	PSH05-51

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: 71 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within Human PABP protein aa 450-550.
Positive control:	TF-1 cell lysate, HeLa cell lysate, A549 cell lysate, MCF7 cell lysate, HepG2 cell lysate, HCT 116 cell lysate, HEK-293 cell lysate, NIH/3T3 cell lysate, COS-1 cell lysate, Vero cell lysate, mouse brain tissue lysate, HeLa, NIH/3T3, human colon tissue, human testis tissue, mouse testis tissue.
Subcellular location:	Cytoplasm, Stress granule, Nucleus, Cell projection, lamellipodium.
Recommended Dilutions: WB IF-Cell IHC-P FC IF-Tissue	1:1,000 1:100 1:1,000 1:1,000 1:200
Uniprot #:	SwissProt: P11940 Human \| P29341 Mouse
Alternative names:	PAB 1 PAB1 PABP 1 PABP-1 PABP1 PABP1_HUMAN PABPC 1 Pabpc1 PABPC2 PABPL1 Poly A binding protein 1 Poly A binding protein cytoplasmic 1 poly(A) binding protein, cytoplasmic 1 poly(A) binding protein, cytoplasmic 2 Poly(A)-binding protein 1 Polyadenylate binding protein 1 Polyadenylate-binding protein 1

Images

Fig1: Western blot analysis of PABP on different lysates with Rabbit anti-PABP antibody (HA722289) at 1/1,000 dilution.

Lane 1: TF-1 cell lysate (20 µg/Lane)
Lane 2: HeLa cell lysate (20 µg/Lane)
Lane 3: A549 cell lysate (20 µg/Lane)
Lane 4: MCF7 cell lysate (20 µg/Lane)
Lane 5: HepG2 cell lysate (20 µg/Lane)
Lane 6: HCT 116 cell lysate (20 µg/Lane)
Lane 7: HEK-293 cell lysate (20 µg/Lane)
Lane 8: NIH/3T3 cell lysate (20 µg/Lane)
Lane 9: COS-1 cell lysate (20 µg/Lane)
Lane 10: Vero cell lysate (20 µg/Lane)
Lane 11: Mouse brain tissue lysate (40 µg/Lane)

Predicted band size: 71 kDa
Observed band size: 71 kDa

Exposure time: 20 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 (HA722289) 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 HeLa cells labeling PABP with Rabbit anti-PABP antibody (HA722289) 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-PABP antibody (HA722289) 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: Immunocytochemistry analysis of NIH/3T3 cells labeling PABP with Rabbit anti-PABP antibody (HA722289) 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-PABP antibody (HA722289) 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-PABP antibody (HA722289) 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 (HA722289) 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 human testis tissue with Rabbit anti-PABP antibody (HA722289) 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 (HA722289) 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: Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-PABP antibody (HA722289) 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 (HA722289) 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.

Fig7: Flow cytometric analysis of HeLa cells labeling PABP.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA722289, 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).

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