HA610206

Claudin 7 Recombinant Mouse Monoclonal Antibody [6-G3-R]

cat.: HA610206

Product Type:	Recombinant Mouse monoclonal IgG1, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P, IF-Tissue, IP
Clonality:	Monoclonal
Clone number:	6-G3-R

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: 22 kDa
Isotype:	IgG1
Immunogen:	Synthetic peptide within Human Claudin 7 aa 162-211 / 211.
Positive control:	SW480 cell lysate, MCF7 cell lysate, LNCaP cell lysate, Caco-2 cell lysate, A431 cell lysate, mouse colon tissue lysate, rat colon tissue lysate, PC-12, human kidney tissue, human colon tissue, mouse colon tissue, rat colon tissue.
Subcellular location:	Cell membrane, Basolateral cell membrane, Cell junction, tight junction.
Recommended Dilutions: WB IF-Cell IHC-P IF-Tissue IP	1:1,000 1:100 1:5,000 1:200-1:1,000 1-2μg/sample
Uniprot #:	SwissProt: O95471 Human \| Q9Z261 Mouse \| Q9Z1L1 Rat
Alternative names:	CEPTR L2 CEPTRL 2 CEPTRL2 Claudin 1 Claudin 9 Claudin-7 Claudin1 Claudin7 Claudin9 CLD7_HUMAN CLDN 7 CLDN-7 CLDN7 Clostridium perfringens enterotoxin receptor like 2 CPETR L2 CPETRL 2 CPETRL2 Hs.84359

Images

Fig1: Western blot analysis of Claudin 7 on different lysates with Mouse anti-Claudin 7 antibody (HA610206) at 1/1,000 dilution.

Lane 1: SW480 cell lysate (20 µg/Lane)
Lane 2: MCF7 cell lysate (20 µg/Lane)
Lane 3: LNCaP cell lysate (20 µg/Lane)
Lane 4: Caco-2 cell lysate (20 µg/Lane)
Lane 5: A431 cell lysate (20 µg/Lane)
Lane 6: Mouse colon tissue lysate (40 µg/Lane)
Lane 7: Rat colon tissue lysate (40 µg/Lane)

Predicted band size: 22 kDa
Observed band size: 20 kDa

Exposure time: 1 minute; 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 (HA610206) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig2: Immunocytochemistry analysis of PC-12 cells labeling Claudin 7 with Mouse anti-Claudin 7 antibody (HA610206) 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 Mouse anti-Claudin 7 antibody (HA610206) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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 (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

	Fig3: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Mouse anti-Claudin 7 antibody (HA610206) 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 (HA610206) 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded human colon tissue with Mouse anti-Claudin 7 antibody (HA610206) 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 (HA610206) 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Mouse anti-Claudin 7 antibody (HA610206) 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 (HA610206) 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.

Fig6: Immunohistochemical analysis of paraffin-embedded rat colon tissue with Mouse anti-Claudin 7 antibody (HA610206) 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 (HA610206) 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.

Fig7: Claudin 7 was immunoprecipitated from 0.2 mg A431 cell lysate with HA610206 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA610206 at 1/2,000 dilution. Anti-Mouse IgG for IP Nano-secondary antibody (NBI02H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: A431 cell lysate (input)
Lane 2: HA610206 IP in A431 cell lysate
Lane 3: Mouse IgG instead of HA610206 in A431 cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 2 minutes; ECL: K1801

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