HA500355

TIP49A Rabbit Polyclonal Antibody

cat.: HA500355

Product Type:	Rabbit polyclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, IF-Cell, FC
Clonality:	Polyclonal

Form:	Liquid
Storage condition:	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:	PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Immunogen affinity purified.
Molecular weight:	Predicted band size: 50 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within Human TIP49A aa 121-320 / 456.
Positive control:	HeLa cell lysate, MCF7 cell lysate, NIH/3T3 cell lysate, C6 cell lysate, Mouse testis tissue lysate, Rat testis tissue lysate, human testis tissue, mouse testis tissue, rat testis tissue, HeLa, NIH/3T3, C6.
Subcellular location:	Cytoplasm, Cytoskeleton, Membrane, Nucleus.
Recommended Dilutions: WB IHC-P IF-Cell FC	1:50,000 1:5,000 1:100 1:5,000
Uniprot #:	SwissProt: Q9Y265 Human \| P60122 Mouse \| P60123 Rat
Alternative names:	49 kDa TATA box binding protein interacting protein 49 kDa TATA box-binding protein-interacting protein 49 kDa TBP interacting protein 49 kDa TBP-interacting protein 54 kDa erythrocyte cytosolic protein ECP-54 ECP54 ERYTHROCYTE CYTOSOLIC PROTEIN, 54-KD INO80 complex subunit H NMP 238 Nuclear matrix protein 238 Pontin 52 PONTIN RuvB like 1 RuvB like AAA ATPase 1 RuvB-like 1 RUVB1_HUMAN RUVBL1 RVB1 TAP54 alpha TAP54-alpha TIP49 TIP49a TIP60 associated protein 54 alpha TIP60-associated protein 54-alpha

Images

Fig1: Western blot analysis of TIP49A on different lysates with Rabbit anti-TIP49A antibody (HA500355) at 1/50,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: MCF7 cell lysate
Lane 3: NIH/3T3 cell lysate
Lane 4: C6 cell lysate
Lane 5: Mouse testis tissue lysate
Lane 6: Rat testis tissue lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 50 kDa
Observed band size: 50 kDa

Exposure time: 2 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 (HA500355) at 1/50,000 dilution was used in primary antibody dilution (K1803) 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: Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-TIP49A antibody (HA500355) at 1/5,000 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 (HA500355) at 1/5,000 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 mouse testis tissue with Rabbit anti-TIP49A antibody (HA500355) at 1/5,000 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 (HA500355) at 1/5,000 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 rat testis tissue with Rabbit anti-TIP49A antibody (HA500355) at 1/5,000 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 (HA500355) at 1/5,000 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: Immunocytochemistry analysis of HeLa cells labeling TIP49A with Rabbit anti-TIP49A antibody (HA500355) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-TIP49A antibody (HA500355) 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 (HA601187, 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: Immunocytochemistry analysis of NIH/3T3 cells labeling TIP49A with Rabbit anti-TIP49A antibody (HA500355) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-TIP49A antibody (HA500355) 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 (HA601187, 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.
	Fig7: Immunocytochemistry analysis of C6 cells labeling TIP49A with Rabbit anti-TIP49A antibody (HA500355) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-TIP49A antibody (HA500355) 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 (HA601187, 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.
	Fig8: Flow cytometric analysis of HeLa cells labeling TIP49A. Cells were fixed and permeabilized. Then stained with the primary antibody (HA500355, 1/5,000) (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).

Fig9: Flow cytometric analysis of NIH/3T3 cells labeling TIP49A.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA500355, 1/5,000) (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).

Fig10: Flow cytometric analysis of C6 cells labeling TIP49A.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA500355, 1/5,000) (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).

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