HA751777

ATRX Recombinant Rabbit Monoclonal Antibody [PSH20-95]

cat.: HA751777

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P, IP
Clonality:	Monoclonal
Clone number:	PSH20-95

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*PBS (pH7.4).
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 283 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within human ATRX aa 2,193-2,492.
Positive control:	SH-SY5Y cell lysate, HeLa cell lysate, MOLT-4 cell lysate, NIH/3T3 cell lysate, C2C12 cell lysate, L6 cell lysate, C6 cell lysate, NRK cell lysate, NR8383 cell lysate, PC-12 cell lysate, human breast carcinoma tissue, human colon tissue, mouse brain tissue, mouse colon tissue, rat brain tissue, rat colon tissue, SH-SY5Y, NIH/3T3.
Subcellular location:	Nucleus, Chromosome, telomere, PML body.
Recommended Dilutions: WB IF-Cell IHC-P IP	1:5,000-1:10,000 1:1,000 1:200-1:1,000 1-2μg/sample
Uniprot #:	SwissProt: P46100 Human \| Q61687 Mouse \| P70486 Rat
Alternative names:	Alpha thalassemia/mental retardation syndrome X linked homolog ATP dependent helicase ATRX ATP-dependent helicase ATRX ATR2 Atrx ATRX_HUMAN DNA dependent ATPase and helicase Helicase 2, X linked MGC2094 MRXHF1 RAD54 RAD54L SFM1 SHS Transcriptional regulator ATRX X linked helicase II X linked nuclear protein X-linked helicase II X-linked nuclear protein XH2 XNP Znf HX Znf-HX

Images

Fig1: Western blot analysis of ATRX on different lysates with Rabbit anti-ATRX antibody (HA751777) at 1/5,000 dilution.

Lane 1: SH-SY5Y cell lysate
Lane 2: U-2 OS cell lysate (negative)
Lane 3: HeLa cell lysate
Lane 4: MOLT-4 cell lysate
Lane 5: NIH/3T3 cell lysate
Lane 6: C2C12 cell lysate
Lane 7: L6 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 283 kDa
Observed band size: 400 kDa

Exposure time: 10 seconds; ECL: K1801;
3-8% 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 (HA751777) at 1/5,000 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: Western blot analysis of ATRX on different lysates with Rabbit anti-ATRX antibody (HA751777) at 1/10,000 dilution.

Lane 1: L6 cell lysate
Lane 2: C6 cell lysate
Lane 3: NRK cell lysate
Lane 4: NR8383 cell lysate
Lane 5: PC-12 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 283 kDa
Observed band size: 400 kDa

Exposure time: 3 minutes; ECL: K1801;
3-8% 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 (HA751777) at 1/10,000 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.

Fig3: Immunocytochemistry analysis of SH-SY5Y cells labeling ATRX with Rabbit anti-ATRX antibody (HA751777) at 1/1,000 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-ATRX antibody (HA751777) at 1/1,000 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.

Fig4: Immunocytochemistry analysis of NIH/3T3 cells labeling ATRX with Rabbit anti-ATRX antibody (HA751777) at 1/1,000 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-ATRX antibody (HA751777) at 1/1,000 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.

	Fig5: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-ATRX antibody (HA751777) at 1/200 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 (HA751777) at 1/200 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 human colon tissue with Rabbit anti-ATRX antibody (HA751777) at 1/200 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 (HA751777) at 1/200 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: Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-ATRX antibody (HA751777) 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 (HA751777) 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.

	Fig8: Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-ATRX antibody (HA751777) at 1/200 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 (HA751777) at 1/200 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.
	Fig9: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-ATRX antibody (HA751777) 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 (HA751777) 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.
	Fig10: Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-ATRX antibody (HA751777) at 1/200 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 (HA751777) at 1/200 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.

Fig11: ATRX was immunoprecipitated from 0.2 mg SH-SY5Y cell lysate with HA751777 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751777 at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: SH-SY5Y cell lysate (input)
Lane 2: HA751777 IP in SH-SY5Y cell lysate
Lane 3: Rabbit IgG instead of HA751777 in SH-SY5Y cell lysate

Blocking/Dilution buffer: primary antibody dilution (K1803)
Exposure time: 20 seconds; ECL: K1801

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