ET1702-54

Calbindin Recombinant Rabbit Monoclonal Antibody [JF05-01]

cat.: ET1702-54

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, mIHC, IHC-Fr
Clonality:	Monoclonal
Clone number:	JF05-01

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. 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: 30 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human Calbindin aa 51-100 / 261.
Positive control:	Mouse brain tissue lysate, rat brain tissue lysate, mouse kidney tissue lysate, rat kidney tissue lysate, human kidney tissue, mouse cerebellum tissue, PC-12, 293T, rat brain tissue, mouse brain tissue, mouse kidney tissue.
Subcellular location:	Cytosol, Nucleus.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P mIHC IHC-Fr	1:1,000-1:5,000 1:50-1:200 1:500 1:50-1:5,000 1:4,000 1:500
Uniprot #:	SwissProt: P05937 Human \| P12658 Mouse \| P07171 Rat
Alternative names:	avian-type CAB27 CALB 1 CALB CALB1 CALB1_HUMAN Calbindin 1 28kDa Calbindin Calbindin D28 D 28K D-28K D28K OTTHUMP00000166027 OTTHUMP00000225441 RTVL H protein Vitamin D dependent calcium binding protein Vitamin D dependent calcium binding protein avian type Vitamin D-dependent calcium-binding protein

Images

Fig1: Fluorescence multiplex immunohistochemical analysis of mouse kidney (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-AQP1 (ET1703-34, Violet), anti-CK18 (ET1603-8, Green) and anti-Calbindin (ET1702-54, Yellow) on kidney. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKit™MH010101, www.luminiris.cn). The section was incubated in three rounds of staining: in the order of ET1703-34 (1/5,000 dilution), ET1603-8 (1/3,000 dilution) and ET1702-54 (1/4,000 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Zeiss Observer 7 Inverted Fluorescence Microscope.

Fig2: Fluorescence multiplex immunohistochemical analysis of human kidney (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-CD31 (M1511-8, Red), anti-E-Cadherin (HA601143, Green), anti-Calbindin (ET1702-54, Magenta) on human kidney. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKit™MH010101, www.luminiris.cn). The section was incubated in three rounds of staining: in the order of M1511-8 (1/1,000 dilution), HA601143 (1/4,000 dilution) and ET1702-54 (1/4,000 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.

	Fig3: Immunofluorescence analysis of frozen mouse cerebellum tissue with Rabbit anti-Calbindin antibody (ET1702-54) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1702-54, green) at 1/500 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
	Fig4: Western blot analysis of Calbindin on different lysates with Rabbit anti-Calbindin antibody (ET1702-54) at 1/5,000 dilution. Lane 1: Mouse brain tissue lysate Lane 2: NIH/3T3 cell lysate (negative) Lane 3: Rat brain tissue lysate Lane 4: C6 cell lysate (negative) Lane 5: Mouse kidney tissue lysate Lane 6: Rat kidney tissue lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 30 kDa Observed band size: 30 kDa Exposure time: 6 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 (ET1702-54) at 1/5,000 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.
	Fig5: Immunofluorescence analysis of paraffin-embedded mouse cerebellum tissue labeling Calbindin with Rabbit anti-Calbindin antibody (ET1702-54) at 1/500 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1702-54, green) at 1/500 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

	Fig6: Immunofluorescence analysis of paraffin-embedded rat cerebellum tissue labeling Calbindin with Rabbit anti-Calbindin antibody (ET1702-54) at 1/500 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1702-54, green) at 1/500 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
	Fig7: Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue with Rabbit anti-Calbindin antibody (ET1702-54) 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 (ET1702-54) 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.
	Fig8: ICC staining Calbindin in PC-12 cells (red). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
	Fig9: ICC staining Calbindin in 293T cells (red). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

	Fig10: Immunohistochemical analysis of paraffin-embedded rat brain tissue using anti-Calbindin antibody. Counter stained with hematoxylin.
	Fig11: Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-Calbindin antibody. Counter stained with hematoxylin.
	Fig12: Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-Calbindin antibody. Counter stained with hematoxylin.
	Fig13: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue using anti-Calbindin antibody. Counter stained with hematoxylin.

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