36-4900

antibody from Invitrogen Antibodies

Targeting: GJA5 CX40

Provider product page for 36-4900

Western blot

Immunoprecipitation

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [25]
Comments [0]
Validations
Western blot [1]
Immunohistochemistry [2]
Other assay [24]

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Product number: 36-4900 - Provider product page
Provider: Invitrogen Antibodies
Product name: Connexin 40 Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Reactivity: Human, Mouse, Rat
Host: Rabbit
Isotype: IgG
Vial size: 100 µg
Concentration: 0.25 mg/mL
Storage: -20°C

GJA5 protein structure - 36-4900 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry analysis of Connexin 40 showing staining in the membrane of paraffin-embedded human heart tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Connexin 40 polyclonal antibody (Product # 36-4900) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Connexin 40 showing staining in the membrane of paraffin-embedded mouse lung tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Connexin 40 polyclonal antibody (Product # 36-4900) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Peptide Competition and Induction: Immunoprecipitates prepared from lysates of Jurkat cells stimulated with TNF-α (1-4, 6) or left unstimulated (5) were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4øC and incubated with 0.75 µg/mL IKKα (pSpS176/180) antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 5, 6), the non-phosphopeptide corresponding to the immunogen (2), a generic phosphoserine containing peptide (3), or, the phosphopeptide immunogen (4). After washing, membranes were incubated with goat F (ab')2 anti-rabbit IgG HRP conjugate (Product # ALI4404) and bands were detected using the Pierce SuperSignal? method. The data show that the peptide corresponding to IKKα (pSpS176/180) blocks the antibody signal, thereby demonstrating the specificity of the antibody. The data also show that the phospho-signal is induced by the addition of TNF-α.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 8 LDL reduces connexin-40 expression in HL-1 myocytes. Effect of increasing doses of LDL (0, 50, 100 and 200 ug/mL) on connexin-40 (Cx40) and connexin-43 (Cx43) mRNA ( A ) or protein ( B ) levels in HL-1 myocyte cultures C Relationship between intracellular cholesteryl ester (CE) and Cx40 and Cx43 protein levels in HL-1 myocyte cultures. Results are shown as mean+-SEM of three independent experiments performed in duplicate. *P

Supportive validation

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Experimental details: Figure 2. Embryonic fourth PAA formation is disrupted by endothelial Hey1 deficiency. A , immunohistochemistry of Pecam1 and Nrp1 revealed defective tubular structure of fourth PAAs ( arrows ) in endothelial cKO ( Hey1 fl/ Delta ;Tek-Cre + ) embryos at E10.5 and E11.5. B , Gja5 immunohistochemistry at E11.5 also showed the impaired endothelial tube formation of fourth PAAs. Expression levels of these endothelial markers remained unchanged. C , expression of a smooth muscle marker Acta2 was almost undetectable in the affected fourth PAA. PAAs are numbered. Note that the staining of different markers is shown using serial sections for E11.5. Scale bars , 100 mum.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 4 sDCI imaging of IDs. Representative 3D sDCI images of en face IDs from murine atria immunolabeled for (A, B) Na V 1.5, Cx40, Cx43, and N-cad, and (C, D) Na V 1.5, beta1, Cx43, and N-cad.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2 Gene and protein expression ( A - C ). Expression levels of CACNA1C , SLC8A1 and the connexin members, GJA5 , GJA1 and GJC1 in HL-1 cells transfected with miR-NC (black columns) or miR-199a+miR-22 (white columns). Normalized to miR-NC ( D - F ). Protein expression levels of Cav1.2, NCX1 and Cx40, normalized to the endogenous control, tubulin ( G ). Representative WB experiments in HL-1 cells transfected with miR-22+199a and miR-NC. All proteins (Cav1.2, NCX1, connexin 40 and tubulin) were blotted on the same gel. This is a representative image of four independent experiments. Numbers within columns represent the number of independent experiments done. * p < 0.05, *** p < 0.0001, n.s. : non-significant.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 8 CX40 mediates TET1s-induced endothelial barrier reinforcement. (A) Heatmap of the top 20 selected upregulated genes by RNA sequencing. (B) RT-qPCR was used to test the mRNA levels of the top 5 upregulated genes from RNA-seq and three hemodynamic-sensitive genes. (C) The CX40 protein expression level was quantified by WB (n=6 per group). (D-L) Stable CX40 -/- p-HUVECs were generated by transfecting human connexin 40-specific CRISPR/Cas9 KO plasmids. Then, TET1s-adenovirus was used to transfect CX40 -/- and CX40 +/+ p-HUVECs to generate CX40 +/+ +NC, CX40 +/+ +OE, CX40 -/- +NC and CX40 -/- +OE p-HUVECs. (D) The fluorescence intensity of the lower chamber medium was tested as described in Fig. 3 C (n>6 per group). (E, H) Immunofluorescence staining for F-actin and VE-cadherin. The green dotted line indicates the intercellular space area. (F-G) Quantitative analysis of single-cell F-actin length and intercellular space area to image E (n>10 per group). (I-K) Quantitative analysis of VE-cadherin discontinuity, intercellular space area and ratio of VE-cadherin in several morphological categories to image H (n>10 per group). All data were presented as the mean +- SD.