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Calbindin Recombinant Rabbit Monoclonal Antibody [JF05-01]

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Specification

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Catalog# ET1702-54

Calbindin Recombinant Rabbit Monoclonal Antibody [JF05-01]

Application

  • WB

  • IF-Tissue

  • IHC-P

  • mIHC

  • IHC-Fr

  • VIVIT

Reactivity

  • Mouse

  • Rat

BSA and Azide free
Predicted reactivity

  • Cynomolgus monkey

  • Pig

Predicted species support after-sales service
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Calbindin Recombinant Rabbit Monoclonal Antibody [JF05-01]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human Calbindin aa 51-100 / 261.

Species Reactivity

Mouse, Rat (Predicted: Cynomolgus monkey, Pig)

Predicted species support after-sales service

Validated Applications

WB, IF-Tissue, IHC-P, mIHC, IHC-Fr, VIVIT

Molecular Weight

Predicted band size: 30 kDa

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.

Conjugation

unconjugated

Clone Number

JF05-01

RRID

AB_3070316

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:5,000

  • IF-Tissue

  • 1:500

  • IHC-P

  • 1:5,000-1:10,000

  • mIHC

  • 1:4,000

  • IHC-Fr

  • 1:2,000

Target

Function

Calbindin 1 is a protein that in humans is encoded by the CALB1 gene. It belongs to the calbindin family of calcium-binding proteins, along with calretinin (CALB2). The protein encoded by this gene is a member of the calcium-binding protein superfamily that includes calmodulin and troponin C. Originally described as a 27 kDa protein, it is now known to be a 28 kDa protein. It contains four active calcium-binding domains, and has two modified domains that are thought to have lost their calcium binding capability. This protein is thought to buffer entry of calcium upon stimulation of glutamate receptors. Depletion of this protein was noted in patients with Huntington disease.

Background References

1. Noble JW et al. Exploring Calbindin-IMPase fusion proteins structure and activity. Biochem Biophys Rep. 2022 May

2. Attig J et al. Human endogenous retrovirus onco-exaptation counters cancer cell senescence through calbindin. J Clin Invest. 2023 Jul

Sequence Similarity

Belongs to the calbindin family.

Subcellular Location

Cytosol, Nucleus.

UNIPROT

SwissProt: P05937 Human

SwissProt: P12658 Mouse

SwissProt: P07171 Rat

Synonyms

avian-type antibody

CAB27 antibody

CALB 1 antibody

CALB antibody

CALB1 antibody

CALB1_HUMAN antibody

Calbindin 1 28kDa antibody

Calbindin antibody

Calbindin D28 antibody

D 28K antibody

Expand

Images

  • Application: IHC-Fr<br /><br />Species: Mouse<br /><br />Site: Cerebellum <br /><br />Sample: Frozen section<br /><br />Antibody concentration: 1/2,000<br /><br />Antigen retrieval: Not required

    Application: IHC-Fr

    Species: Mouse

    Site: Cerebellum

    Sample: Frozen section

    Antibody concentration: 1/2,000

    Antigen retrieval: Not required

  • Fluorescence multiplex immunohistochemical analysis of mouse kidney (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-AQP1 (<a href="/products/ET1703-34" style="font-weight: bold;text-decoration: underline;">ET1703-34</a>, Violet), anti-CK18 (<a href="/products/ET1603-8" style="font-weight: bold;text-decoration: underline;">ET1603-8</a>, Green) and anti-Calbindin (<a href="/products/ET1702-54" style="font-weight: bold;text-decoration: underline;">ET1702-54</a>, 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&trade;MH010101, www.luminiris.cn). The section was incubated in three rounds of staining: in the order of <a href="/products/ET1703-34" style="font-weight: bold;text-decoration: underline;">ET1703-34</a> (1/5,000 dilution), <a href="/products/ET1603-8" style="font-weight: bold;text-decoration: underline;">ET1603-8</a> (1/3,000 dilution) and <a href="/products/ET1702-54" style="font-weight: bold;text-decoration: underline;">ET1702-54</a> (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.

    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.

  • Fluorescence multiplex immunohistochemical analysis of human kidney (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-CD31 (<a href="/products/M1511-8" style="font-weight: bold;text-decoration: underline;">M1511-8</a>, Red), anti-E-Cadherin (<a href="/products/HA601143" style="font-weight: bold;text-decoration: underline;">HA601143</a>, Green), anti-Calbindin (<a href="/products/ET1702-54" style="font-weight: bold;text-decoration: underline;">ET1702-54</a>, 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&trade;MH010101, www.luminiris.cn). The section was incubated in three rounds of staining: in the order of <a href="/products/M1511-8" style="font-weight: bold;text-decoration: underline;">M1511-8</a> (1/1,000 dilution), <a href="/products/HA601143" style="font-weight: bold;text-decoration: underline;">HA601143</a> (1/4,000 dilution) and <a href="/products/ET1702-54" style="font-weight: bold;text-decoration: underline;">ET1702-54</a> (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.

    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.

  • Application: IF-tissue<br /><br />Species: Mouse<br /><br />Site: Cerebellum<br /><br />Sample: Paraffin-embedded section<br /><br />Antibody concentration: 1/500

    Application: IF-tissue

    Species: Mouse

    Site: Cerebellum

    Sample: Paraffin-embedded section

    Antibody concentration: 1/500

  • Application: IF-tissue<br /><br />Species: Rat<br /><br />Site: Cerebellum<br /><br />Sample: Paraffin-embedded section<br /><br />Antibody concentration: 1/500

    Application: IF-tissue

    Species: Rat

    Site: Cerebellum

    Sample: Paraffin-embedded section

    Antibody concentration: 1/500

  • Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue with Rabbit anti-Calbindin antibody (<a href="/products/ET1702-54" style="font-weight: bold;text-decoration: underline;">ET1702-54</a>) at 1/10,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1702-54" style="font-weight: bold;text-decoration: underline;">ET1702-54</a>) at 1/10,000 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.

    Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue with Rabbit anti-Calbindin antibody (ET1702-54) at 1/10,000 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 1/10,000 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.

  • Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue with Rabbit anti-Calbindin antibody (<a href="/products/ET1702-54" style="font-weight: bold;text-decoration: underline;">ET1702-54</a>) at 1/10,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1702-54" style="font-weight: bold;text-decoration: underline;">ET1702-54</a>) at 1/10,000 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.

    Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue with Rabbit anti-Calbindin antibody (ET1702-54) at 1/10,000 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 1/10,000 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.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of Calbindin on different lysates with Rabbit anti-Calbindin antibody (<a href="/products/ET1702-54" style="font-weight: bold;text-decoration: underline;">ET1702-54</a>) at 1/5,000 dilution.<br /><br />Lane 1: Mouse brain tissue lysate<br />Lane 2: NIH/3T3 cell lysate (negative)<br />Lane 3: Rat brain tissue lysate<br />Lane 4: C6 cell lysate (negative)<br />Lane 5: Mouse kidney tissue lysate<br />Lane 6: Rat kidney tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 30 kDa<br />Observed band size: 30 kDa<br /><br />Exposure time: 6 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1702-54" style="font-weight: bold;text-decoration: underline;">ET1702-54</a>) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Relative expression (RE)

    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.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation