ET1703-14

Laminin beta 1 Recombinant Rabbit Monoclonal Antibody [JM099-03]

cat.: ET1703-14

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, FC, mIHC
Clonality:	Monoclonal
Clone number:	JM099-03

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: 198 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human Laminin beta 1 aa 1,721-1,769 / 1,786.
Positive control:	HepG2 cell lysate, C6 cell lysate, mouse stomach tissue lysate, rat heart tissue lysate, human kidney tissue, mouse colon tissue, mouse prostate tissue, mouse kidney tissue, mouse small intestine tissue, A431.
Subcellular location:	Secreted.
Recommended Dilutions: WB IHC-P FC mIHC	1:2,000 1:50-1:200 1:50-1:100 1:1,000
Uniprot #:	SwissProt: P07942 Human \| P02469 Mouse Entrez Gene: 298941 Rat
Alternative names:	CLM Cutis laxa with marfanoid phenotype LAM B1 LAMB 1 LAMB1 LAMB1_HUMAN Laminin B1 Laminin B1 chain Laminin beta 1 chain Laminin beta 1 chain precursor Laminin beta1 Laminin subunit beta 1 Laminin subunit beta-1 Laminin-1 subunit beta Laminin-10 subunit beta Laminin-12 subunit beta Laminin-2 subunit beta Laminin-6 subunit beta Laminin-8 subunit beta LIS5 MGC142015

Images

Fig1: Western blot analysis of Laminin beta 1 on different lysates with Rabbit anti-Laminin beta 1 antibody (ET1703-14) at 1/2,000 dilution.

Lane 1: HepG2 cell lysate (15 µg/Lane)
Lane 2: C6 cell lysate (15 µg/Lane)
Lane 3: Mouse stomach tissue lysate (20 µg/Lane)
Lane 4: Rat heart tissue lysate (20 µg/Lane)

Predicted band size: 198 kDa
Observed band size: 250 kDa

Exposure time: 3 minutes 10 seconds;

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 (ET1703-14) 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: Fluorescence multiplex immunohistochemical analysis of mouse kidney (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-NPHS2 (ET7107-34, Red), anti-AQP1 (ET1703-34, Green), anti-Laminin beta 1 (ET1703-14, Cyan) and anti-aSMA (ET1607-53, Magenta) 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 four rounds of staining: in the order of ET7107-34 (1/1,000 dilution), ET1703-34 (1/5,000 dilution), ET1703-14 (1/1,000 dilution) and ET1607-53 (1/10,000 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: Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-Laminin beta 1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1703-14, 1/50) for 30 minutes 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 mouse colon tissue using anti-Laminin beta 1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1703-14, 1/50) for 30 minutes 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 prostate tissue using anti-Laminin beta 1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1703-14, 1/50) for 30 minutes 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 kidney tissue using anti-Laminin beta 1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1703-14, 1/50) for 30 minutes 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 small intestine tissue using anti-Laminin beta 1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1703-14, 1/50) for 30 minutes 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 Laminin beta 1 was done on A431 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1703-14, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.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”.