ER62856

CA12 Rabbit Polyclonal Antibody

cat.: ER62856

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). Store at -20℃ long term.
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: 39 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within human CA12 aa 1-301.
Positive control:	A549 cell lysate, MCF7 cell lysate, HeLa cell lysate, U-87 MG cell lysate, NIH:OVCAR-3 cell lysate, Mouse colon tissue lysate, Rat colon tissue lysate, A549, human kidney tissue, human colon tissue, mouse colon tissue, rat colon tissue.
Subcellular location:	Membrane.
Recommended Dilutions: WB IHC-P IF-Cell FC	1:2,000 1:1,000 1:100 1:1,000
Uniprot #:	SwissProt: O43570 Human \| Q8CI85 Mouse
Alternative names:	CA 12 CA XII CA-XII CA12 CAH12_HUMAN Carbonate dehydratase XII Carbonic anhydrase 12 Carbonic anhydrase XII Carbonic dehydratase CAXII FLJ20151 HsT18816 T18816 Tumor antigen HOM RCC 3.1.3 Tumor antigen HOM-RCC-3.1.3

Images

Fig1: Western blot analysis of CA12 on different lysates with Rabbit anti-CA12 antibody (ER62856) at 1/2,000 dilution.

Lane 1: A549 cell lysate (15 µg/Lane)
Lane 2: MCF7 cell lysate (15 µg/Lane)
Lane 3: HeLa cell lysate (15 µg/Lane)
Lane 4: U-87 MG cell lysate (15 µg/Lane)
Lane 5: NIH:OVCAR-3 cell lysate (15 µg/Lane)
Lane 6: Mouse colon tissue lysate (20 µg/Lane)
Lane 7: Rat colon tissue lysate (20 µg/Lane)

Predicted band size: 40 kDa
Observed band size: 45 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 (ER62856) 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: Immunocytochemistry analysis of A549 cells labeling CA12 with Rabbit anti-CA12 antibody (ER62856) 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-CA12 antibody (ER62856) 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: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-CA12 antibody (ER62856) 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 (ER62856) 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 human colon tissue with Rabbit anti-CA12 antibody (ER62856) 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 (ER62856) 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 mouse colon tissue with Rabbit anti-CA12 antibody (ER62856) 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 (ER62856) 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 rat colon tissue with Rabbit anti-CA12 antibody (ER62856) 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 (ER62856) 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 A549 cells labeling CA12.

Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (ER62856, 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”.