ET7109-16

IMPDH2 Recombinant Rabbit Monoclonal Antibody [JE40-87]

cat.: ET7109-16

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, FC, IP
Clonality:	Monoclonal
Clone number:	JE40-87

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 56 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within Human IMPDH2 aa 80-320 / 514.
Positive control:	A431 cell lysate, Daudi cell lysate, HeLa cell lysate, K-562 cell lysate, Mouse spleen tissue lysate, Rat spleen tissue lysate, K-562, human colon tissue, human tonsils tissue, mouse spleen tissue, rat spleen tissue.
Subcellular location:	Cytoplasm, Nucleus, cytosol.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P FC IP	1:1,000-1:2,000 1:100 1:50-1:200 1:200-1:1,000 1:1,000 1-2μg/sample
Uniprot #:	SwissProt: P12268 Human \| P24547 Mouse \| E9PU28 Rat
Alternative names:	IMDH2_HUMAN IMP (inosine monophosphate) dehydrogenase 2 IMP dehydrogenase 2 IMP oxireductase 2 IMPD 2 IMPD2 IMPDH 2 IMPDH II IMPDH-II Impdh2 Inosine 5' monophosphate dehydrogenase 2 Inosine 5' phosphate dehydrogenase 2 Inosine monophosphate dehydrogenase type II Inosine-5'-monophosphate dehydrogenase 2

Images

Fig1: Western blot analysis of IMPDH2 on different lysates with Rabbit anti-IMPDH2 antibody (ET7109-16) at 1/2,000 dilution.

Lane 1: A431 cell lysate
Lane 2: Daudi cell lysate
Lane 3: HeLa cell lysate
Lane 4: K-562 cell lysate
Lane 5: Mouse spleen tissue lysate
Lane 6: Rat spleen tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 56 kDa
Observed band size: 56 kDa

Exposure time: 25 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 (ET7109-16) at 1/2,000 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: Western blot analysis of IMPDH2 on different lysates with Rabbit anti-IMPDH2 antibody (ET7109-16) at 1/1,000 dilution.

Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-IMPDH2 KD cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 56 kDa
Observed band size: 56 kDa

Exposure time: 9 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 (ET7109-16) at 1/1,000 dilution was used in 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 K-562 cells labeling IMPDH2 with Rabbit anti-IMPDH2 antibody (ET7109-16) 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-IMPDH2 antibody (ET7109-16) 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-IMPDH2 antibody (ET7109-16) 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 (ET7109-16) 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 tonsils tissue with Rabbit anti-IMPDH2 antibody (ET7109-16) 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 (ET7109-16) 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 spleen tissue with Rabbit anti-IMPDH2 antibody (ET7109-16) 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 (ET7109-16) 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: Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-IMPDH2 antibody (ET7109-16) 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 (ET7109-16) 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.
	Fig8: Flow cytometric analysis of K-562 cells labeling IMPDH2. Cells were fixed and permeabilized. Then stained with the primary antibody (ET7109-16, 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).
	Fig9: IMPDH2 was immunoprecipitated from 0.2 mg K-562 cell lysate with ET7109-16 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET7109-16 at 1/2,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: K-562 cell lysate (input) Lane 2: ET7109-16 IP in K-562 cell lysate Lane 3: Rabbit IgG instead of ET7109-16 in K-562 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 9 seconds; ECL: K1801

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